• Korean Journal of Food Science and Technology
• /
• v.42 no.3
• /
• pp.287-291
• /
• 2010

In this study, a simple and rapid pre-treatment method based on liquid extraction was applied for the simultaneous determination of three macrolides (spiramycin, tylosin, and tilmicosin) residues. In these studies, the stock farm products was used as a matrix sample. When the liquid extraction method was compared with the solid phase extraction (SPE) method, the former showed higher recovery percentages and simpler steps than the latter. The macrolids were separated using a reverse-phase C18 ($250\;mm{\times}4.6\;mm$, $5\;{\mu}m$) column and a gradient elution with mobile phases consisting of phosphate buffer (pH 2.5) and acetonitrile. Tylosin and tilmicosin were detected at 288 nm and spiramycin was detected at 232 nm. The average recovery percentage ranged between 83.0-90.2% for samples spiked with the three macrolids at 50 and 100 ng/g The validation results showed that the limit of detection (7 (spiramycin), 12 (tilmiconsin), 12 (tylosin) ng/g)) was under the regulatory tolerances and the linearity from calibration curves was satisfactory for determining the multi-residue of three macrolids in farm products. Monitoring samples were collected at the main cities in Korea as Seoul, Busan, Deajeon, Incheon, Deagu, and Gwangju. Microlide antibiotics were not detected in most samples.

• Korean Journal of Poultry Science
• /
• v.36 no.2
• /
• pp.177-186
• /
• 2009

It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

• Journal of Life Science
• /
• v.21 no.5
• /
• pp.631-646
• /
• 2011

Mesenchymal stem cells (MSC) are multipotent and can be isolated from diverse human tissues including bone marrow, fat, placenta, dental pulp, synovium, tonsil, and the thymus. They function as regulators of tissue homeostasis. Because of their various advantages such as plasticity, easy isolation and manipulation, chemotaxis to cancer, and immune regulatory function, MSCs have been considered to be a potent cell source for regenerative medicine, cancer treatment and other cell based therapy such as GVHD. However, relating to its supportive feature for surrounding cell and tissue, it has been frequently reported that MSCs accelerate tumor growth by modulating cancer microenvironment through promoting angiogenesis, secreting growth factors, and suppressing anti-tumorigenic immune reaction. Thus, clinical application of MSCs has been limited. To understand the underlying mechanism which modulates MSCs to function as tumor supportive cells, we co-cultured human adipose tissue derived mesenchymal stem cells (ASC) with cancer cell lines H460 and U87MG. Then, expression data of ASCs co-cultured with cancer cells and cultured alone were obtained via microarray. Comparative expression analysis was carried out using DAVID (Database for Annotation, Visualization and Integrated Discovery) and PANTHER (Protein ANalysis THrough Evolutionary Relationships) in divers aspects including biological process, molecular function, cellular component, protein class, disease, tissue expression, and signal pathway. We found that cancer cells alter the expression profile of MSCs to cancer associated fibroblast like cells by modulating its energy metabolism, stemness, cell structure components, and paracrine effect in a variety of levels. These findings will improve the clinical efficacy and safety of MSCs based cell therapy.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.43 no.10
• /
• pp.1477-1483
• /
• 2014

This study investigated the anti-obesity effects of African mango (Irvingia gabonesis, IGOB $131^{TM}$) extract in leptin-deficient obese mice. Experimental groups were treated with two different doses of IGOB $131^{TM}$ (1% and 2% in each AIN93G supplement) for 8 weeks. Treatment of obese mice with both low and high dose of IGOB $131^{TM}$ significantly reduced body weight gain by 10.9% and 13.3%, respectively, compared to control obese mice. Subcutaneous adipose tissue weight of mice was significantly reduced by 18% by low-dose and 23% by high-dose supplementation. This result was supported by micro-CT analysis around the abdominal regions of mice, indicating that the adipose tissue area and volume were significantly reduced by treatment with IGOB $131^{TM}$. Serum levels of triglycerides in the low- and high-dose groups were reduced by 36.5% and 43.8%, respectively, upon treatment with IGOB $131^{TM}$, whereas total cholesterol levels were reduced by 31.8% and 35.4%. Interestingly, the serum LDL level decreased upon treatment with IGOB $131^{TM}$ while the serum level of HDL dramatically increased upon high-dose treatment with IGOB $131^{TM}$, resulting in a significant reduction in the LDL to HDL ratio of 59.2%. These results were supported by the expression levels of enzymes and proteins related to lipid metabolism assessed by real-time PCR. There was a significant increase of in adiponectin expression as well as significant decreases in the expression of FAS, LPL, and lipid regulatory transcription factors such as PPAR-${\gamma}$, C/EBP, and SREBP upon both low- and high-dose IGOB $131^{TM}$ treatment. However, there was no statistical difference between low- and high-dose treatments. These results suggest that IGOB $131^{TM}$ is able to regulate the serum lipid profiles by reducing triglyceride and increasing HDL levels as well as regulate expression of lipid metabolic factors, resulting in reduction of a weight gain in leptin-deficient obese mice.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.51-52
• /
• 2017

"Plant genetic resources for food and agriculture" means any genetic material of plant origin of actual or potential value for food and agriculture. "Genetic material" means any material of plant origin, including reproductive and vegetative propagating material, containing functional units of heredity. (Internal Treaty on Plant Genetic Resources for Food and Agriculture, ITPGRFA). The "Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable Sharing of Benefits Arising from their Utilization (ABS) to the Convention on Biological Diversity (shortly Nagoya Protocol)" is a supplementary agreement to the Convention on Biological Diversity. It provides a transparent legal framework for the effective implementation of one of the three objectives of the CBD: the fair and equitable sharing of benefits arising out of the utilization of genetic resources. The Nagoya Protocol on ABS was adopted on 29 October 2010 in Nagoya, Japan and entered into force on 12 October 2014, 90 days after the deposit of the fiftieth instrument of ratification. Its objective is the fair and equitable sharing of benefits arising from the utilization of genetic resources, thereby contributing to the conservation and sustainable use of biodiversity. The Nagoya Protocol will create greater legal certainty and transparency for both providers and users of genetic resources by; (a) Establishing more predictable conditions for access to genetic resources and (b) Helping to ensure benefit-sharing when genetic resources leave the country providing the genetic resources. By helping to ensure benefit-sharing, the Nagoya Protocol creates incentives to conserve and sustainably use genetic resources, and therefore enhances the contribution of biodiversity to development and human well-being. The Nagoya Protocol's success will require effective implementation at the domestic level. A range of tools and mechanisms provided by the Nagoya Protocol will assist contracting Parties including; (a) Establishing national focal points (NFPs) and competent national authorities (CNAs) to serve as contact points for information, grant access or cooperate on issues of compliance, (b) An Access and Benefit-sharing Clearing-House to share information, such as domestic regulatory ABS requirements or information on NFPs and CNAs, (c) Capacity-building to support key aspects of implementation. Based on a country's self-assessment of national needs and priorities, this can include capacity to develop domestic ABS legislation to implement the Nagoya Protocol, to negotiate MAT and to develop in-country research capability and institutions, (d) Awareness-raising, (e) Technology Transfer, (f) Targeted financial support for capacity-building and development initiatives through the Nagoya Protocol's financial mechanism, the Global Environment Facility (GEF) (Nagoya Protocol). The Rural Development Administration (RDA) leading to conduct management agricultural genetic resources following the 'ACT ON THE PRESERVATION, MANAGEMENT AND USE OF AGRO-FISHERY BIO-RESOURCES' established on 2007. According to $2^{nd}$ clause of Article 14 (Designation, Operation, etc. of Agencies Responsible for Agro-Fishery Bioresources) of the act, the duties endowed are, (a) Matters concerning securing, preservation, management, and use of agro-fishery bioresources; (b) Establishment of an integrated information system for agro-fishery bioresources; (c) Matters concerning medium and long-term preservation of, and research on, agro-fishery bioresources; (d) Matters concerning international cooperation for agro-fishery bioresources and other relevant matters. As the result the RDA manage about 246,000 accessions of plant genetic resources under the national management system at the end of 2016.

• Journal of Life Science
• /
• v.21 no.3
• /
• pp.435-443
• /
• 2011

The purpose of this study is to identify the effects of exercise training [ET, 10~18 m/min (speed), 20~30 min (exercise duration)/a day for 5 day/wk, 6 wk) on PGC-$1{\alpha}$, GLUT-1, Tfam, Cu,Zn-SOD and Mn-SOD proteins in brain of STZ-induced diabetic rats. The male Sprague-Dawley (SD) rats were single-injected intraperitoneally with 50mg/kg of streptozotocin (STZ) to produce STZ-induced diabetic rats. Rats were divided into 3 experimental groups with 8 rats in each group, as follows: (1) non-STZ group (n=8), (2) STZ-CON group (n=8), (3) STZ-EXE group (n=8). The results of this study suggest that i) serum glucose level was significantly reduced in STZ-EXE group compared with STZ-CON group (p<0.05), ii) PGC-$1{\alpha}$ (p<0.001), mtPGC-$1{\alpha}$ (p<0.001), GLUT-1 (p<0.001), and mtTfam (p<0.001) proteins in brain of STZ-induced diabetic rats were significantly increased in STZ-EXE group compared with STZ-CON group, iii) Cu,Zn-SOD (p<0.001) and Mn-SOD (p<0.01) proteins in the STZ-induced diabetic rats were significantly increased in STZ-EXE group compared with STZ-CON group. In conclusion, the findings of the present study reveal that treadmill exercise training increases brain GLUT-1 protein level possibly through up-regulation of PGC-$1{\alpha}$ and Tfam proteins which represent key regulatory components of stimulation of brain mitochondrial biogenesis. In addition, treadmill exercise training may prevent oxidative stress by up-regulation of Cu,Zn-SOD and Mn-SOD proteins in the STZ-induced diabetic rats.

• Journal of Life Science
• /
• v.27 no.2
• /
• pp.155-163
• /
• 2017

Mori Folium, the leaf of Morus alba, is a traditional medicinal herb that shows various pharmacological activities such as antiinflammatory, antidiabetic, antimelanogenesis, antioxidant, antibacterial, antiallergic, and immunomodulatory activities. However, the mechanisms of their inhibitory effects on adipocyte differentiation and adipogenesis remain poorly understood. In the present study, we investigated the inhibition of adipocyte differentiation and adipogenesis by ethanol extracts of Mori Folium (EEMF) in 3T3-L1 preadipocytes. Treatment with EEMF suppressed the terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in the lipid droplet number and lipid content through Oil Red O staining. EEMF significantly reduced the accumulation of cellular triglyceride, which is associated with a significant inhibition of pro-adipogenic transcription factors, including sterol regulatory element-binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR{\gamma}$), and CCAAT/enhancer-binding proteins ${\alpha}$ ($C/EBP{\alpha}$) and ${\beta}$ ($C/EBP{\beta}$). In addition, EEMF potentially downregulated the expression of adipocyte-specific genes, including adipocyte fatty acid binding protein (aP2) and leptin. Furthermore, EEMF treatment effectively increased the phosphorylation of the AMP-activated protein kinase (AMPK) and acetyl CoA carboxylase (ACC); however, treatment with a potent inhibitor of AMPK, compound C, significantly restored the EEMF-induced inhibition of pro-adipogenic transcription factors and adipocyte-specific genes. These results together indicate that EEMF has preeminent effects on the inhibition of adipogenesis through the AMPK signaling pathway, and further studies will be needed to identify the active compounds in Mori Folium.

• Journal of Life Science
• /
• v.26 no.9
• /
• pp.991-998
• /
• 2016

NF-κB acts as a critical transcription factor for the survival of cells via the induction of antiapoptotic genes. Constitutive activation of NF-κB in many types of solid tumors suggests that the inhibition of NF-κB might prevent or inhibit tumorigenesis. Although a number of studies demonstrated that Hsp70 regulated NF-κB activity, the exact mechanism is not clear. This study investigated the functional relationship of Hsp70 and IKKγ in the regulation of NF-κB activation using expression plasmids of components of the IKK complex. Wild-type and deletion mutants of IKKγ were expressed together with Hsp70, and the combined regulatory effect of Hsp70 and IKKγ on NF-κB activation was assayed. Hsp70 suppressed the activation of NF-κB in a reporter plasmid assay. Hsp70 also suppressed the phosphorylation and degradation of IκBα. The suppressive effect of Hsp70 on NF-κB activation was synergistically elevated by IKKγ. The N-terminal IKKβ binding site, C-terminal leucine zipper, and zinc finger domains of IKKγ were not necessary for the suppressive effect. Furthermore, Hsp70 and IKKγ synergistically suppressed the induction of COX-2 expression by lipopolysaccharides in RAW264.7 cells. These results suggest that overexpression of Hsp70 and IKKγ may be a strategic method for inhibition of NF-κB and related diseases.

• Clinical and Experimental Reproductive Medicine
• /
• v.34 no.2
• /
• pp.95-106
• /
• 2007

Objective: We previously identified differentially expressed genes (DEGs) between germinal vesicle (GV) and metaphase II (MII) mouse oocyte. The present study was accomplished as a preliminary study to elucidate the role of ribose 5-phosphate isomerase A (Rpia), the essential enzyme of the pentose phosphate pathway (PPP), in oocyte maturation. We observed expression of Rpia in the mouse and porcine oocytes. Methods: Expression pattern of the 11 MII-selective DEGs in various tissues was evaluated using RT-PCR and selected 4 genes highly expressed in the ovary. According to the oocyte-selective expression profile, we selected Rpia as a target for this study. We identified the porcine Rpia sequence using EST clustering technique, since it is not yet registered in public databases. Results: The extended porcine Rpia nucleotide sequence was submitted and registered to GenBank (accession number EF213106). We prepared primers for porcine Rpia according to this sequence. In contrast to the oocyte-specific expression in the mouse, Rpia was expressed in porcine cumulus and granulosa cells as well as in oocytes. Conclusion: This is the first report on the characterization of the Rpia gene in the mouse and porcine ovarian cells. Results of the present study suggest that the mouse and porcine COCs employ different mechanism of glucose metabolism. Therefore, the different metabolic pathways during in vitro oocyte maturation (IVM) in different species may lead different maturation rates. It is required to study further regarding the role of Rpia in glucose metabolism of oocytes and follicular cell fore exploring the regulatory mechanism of oocyte maturation as well as for finding the finest culture conditions for in vitro maturation.

• Journal of Life Science
• /
• v.20 no.7
• /
• pp.1054-1065
• /
• 2010

SH21B is a natural composition composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). In our previous study, we reported that SH21B inhibited adipogenesis and fat accumulation in 3T3-L1 cells through modulation of various regulators in the adipogenesis pathway. The aim of this study was to analyze the transcriptome profiles for the anti-adipogenic effects of SH21B in 3T3-L1 cells. Total RNAs from SH21B-treated 3T3-L1 cells were reverse-transcribed into cDNAs and hybridized to Affymetrix Mouse Gene 1.0 ST array. From microarray analyses, we identified 2,568 genes of which expressions were changed more than two-fold by SH21B, and the clustering analyses of these genes resulted in 9 clusters. Three clusters among the 9 showed down-regulation by SH21B (cluster 4, cluster 6 and cluster 9), and two clusters showed up-regulation by SH21B (cluster 7 and cluster 8) during the adipogenesis of 3T3-L1 cells. It was found that many genes related to cell proliferation and adipogenesis were included in these clusters. Clusters 4, 6 and 9 included genes which were related with adipogenesis induction and cell cycle arrest. Clusters 7 and 8 included genes related to cell proliferation as well as adipogenesis inhibition. These results suggest that the mechanisms of the anti-adipogenic effects of SH21B may be the modulation of genes involved in cell proliferation and adipogenesis.