• Asian-Australasian Journal of Animal Sciences
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• v.32 no.11
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• pp.1789-1800
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• 2019

Objective: Although alveolar macrophages play a key role in the respiratory immunity of livestock, studies on the mechanism of differentiation and survival of alveolar macrophages are lacking. Therefore, we undertook to investigate changes in the lipid metabolism and survival rate, using 3D4/31 macrophages and Dudleya brittonii which has been used as a traditional asthma treatment. Methods: 3D4/31 macrophages were used as the in vitro porcine alveolar macrophages model. The cells were activated by exposure to phorbol 12-myristate 13-acetate (PMA). Dudleya brittonii extraction was performed with distilled water. For evaluating the cell survival rate, we performed the water-soluble tetrazolium salt cell viability assay and growth curve analysis. To confirm cell death, cell cycle and intracellular reactive oxygen species (ROS) levels were measured using flow cytometric analysis by applying fluorescence dye dichlorofluorescein diacetate and propidium iodide. Furthermore, we also evaluated cellular lipid accumulation with oil red O staining, and fatty acid synthesis related genes expression levels using quantitative polymerase chain reaction (qPCR) with SYBR green dye. Glycolysis, fatty acid oxidation, and tricarboxylic acid (TCA) cycle related gene expression levels were measured using qPCR after exposure to Dudleya brittonii extract (DB) for 12 h. Results: The ROS production and cell death were induced by PMA treatment, and exposure to DB reduced the PMA induced downregulation of cell survival. The PMA and DB treatments upregulated the lipid accumulation, with corresponding increase in the acetyl-CoA carboxylase alpha, fatty acid synthase mRNA expressions. DB-PMA co-treatment reduced the glycolysis genes expression, but increased the expressions of fatty acid oxidation and TCA cycle genes. Conclusion: This study provides new insights and directions for further research relating to the immunity of porcine respiratory system, by employing a model based on alveolar macrophages and natural materials.

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.13-19
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• 2006

To search of new porcine pheromonal odorants for biostimulation control system technologies to improve reproductive efficiency in livestock species, the comparative molecular field analysis (CoMFA) for binding affinity constant $(p(Od)_{50})$ between porcine odorant binding protein (pOBP) and ligands of odorant 2-(cyclohexyloxy) tetrahydrofurane derivatives as substrate molecule was conducted and discussed. In the optimized CoMFA model AIV with chirality $(C_1'(R),\;C_2(S))$ in substrate molecule and atom based fit alignment (A) of odorants, the statistical results showed the best predictability of the binding affinities $(p(Od)_{50})$ based on the LOO cross-validated value $r^2_{cv}.\;(q^2=0.886)$ and non-cross-validated conventional coefficient $(r^2_{ncv}.=0.984)$. the binding affinity constants exhibited a good correlation with steric (40.8%), electrostatic (14.6%) and hydrophobic (44.6%) factors of the substrate molecules. from the analytical results of the contour maps, which may give us some valuable informations to the modification of odorants for effective binding affinity.

• Korean Journal of Agricultural Science
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• v.43 no.1
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• pp.52-60
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• 2016

In animal reproduction, the quality of oocytes and embryos has been evaluated by the expression of specific molecules. Follistatin (FST), which was isolated from follicular fluid, binds and bio-neutralizes the TGF-${\beta}$ superfamily members. Previous studies using the bovine model showed FST could be an important molecular determinant of embryo developmental competence. However, the effect of FST treatment on porcine embryo developmental competence has not been established. In this study, the effect of exogenous FST on porcine embryo developmental competence was investigated during in vitro culture. FST (10 ng/ml) treatment induced a significant decrease in the rate of cell arrest at the 4-cell stage. The expression levels of DNA-methyltransferase 1 (DNMT1), histone deacetylase 1 (HDAC1), and histone deacetylase 2 (HDAC2) were decreased in 4-cell stage embryos. FST treatment also resulted in significant improvements in developmental competence of embryos in terms of blastocyst formation rate and OCT-4 mRNA levels, the latter being related to pluripotency. In conclusion, during in vitro culture, FST treatment significantly ameliorated 4-cell block during embryonic development and improved embryo developmental competence. Therefore, FST treatment may potentially have a functional role in porcine embryogenesis that is broadly applicable to enhance in vitro embryo development.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.3
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• pp.215-222
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• 2020

Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.

• BMB Reports
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• v.34 no.6
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• pp.559-565
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• 2001

Protein carboxyl O-methyltransferase (E.C.2.1.1.24) may play a role in the repair of aged protein that is spontaneously incorporated with isoaspartyl residues. The porcine brain carboxyl O-methyltransferase was cloned in the pET32 vector, and overexpressed in E.coh (BL21) that harbors pETPCMT, which encodes 227 amino acids, including tagging proteins at the N-terminus. The protein sequence of the cloned porcine brain PCMT (r-pbPCMT) shares a 98% identity with that of human erythrocyte PCMT and rat brain PCMT. It is 100% identical with that of bovine brain. The r-pbPCMT was purified using Ni-NTA affinity chromatography and digested by enterokinase in order to remove the protein tags. Then Superdex 75HR gel filtration chromatography was performed. The r-pbPCMT exhibited similar in vitro substrate specificities with the PCMT that was purified from porcine brain. The molecular weight of the enzyme was estimated to be 24.5 kDa on the SDS polyacrylamide gel electrophoresis. The $K_m$ value was $1.1{\times}10^{-7}\;M$ for S-adenosyl-L-methionine. S-adnosyl-L-homocysteine was a competitive type of inhibitor with the $K_i$ value of $1.38{\times}10^{-4}\;M$. The enzyme has optimal activity at pH 6.0 and $37^{\circ}C$. These results indicate that the expressed enzyme is functionally similar to the natural protein. It also suggests that it may be a suitable model to further understand the function of the mammalian enzyme.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.10
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• pp.1499-1512
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• 2014

The purposes of this study are to assess pig farmers' preference for highly pathogenic porcine reproductive and respiratory syndrome (PRRS) vaccine, and estimate the cost and benefit of PRRS vaccination in Vietnam. This study employed an integrated epidemiological and economic analysis which combined susceptible-infectious-recovered (SIR) model, choice experiment (CE) and cost-benefit analysis (CBA) together. The result of SIR model showed the basic reproduction number ($R_0$) of PRRS transmission in this study is 1.3, consequently, the optimal vaccination percentage is 26%. The results of CE in this study indicate that Vietnam pig farmers are showing a high preference for the PRRS vaccine. However, their mean willingness to pay is lower than the potential cost of PRRS vaccine. It can be considered to be one of the reasons that the PRRS vaccination ratio is still low in Vietnam. The results of CBA specified from the whole society's point of view (Social perspective), the benefits of PRRS vaccination are 2.3 to 4.5 times larger than the costs. To support policy making for increasing the PRRS vaccination proportion, this study indicates two ways to increase the vaccination proportion: i) decrease vaccine price by providing a subsidy, ii) provide compensation of culling only for PRRS vaccinated pigs.

• Medical Lasers
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• v.8 no.2
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• pp.64-73
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• 2019

Background and Objectives The effectiveness of many physiotherapy modalities in reducing subcutaneous fat has been investigated in numerous previous studies. However, to the best of our knowledge, there have been no attempts to determine the effectiveness of physiotherapy modalities in body contouring. The present report determined the effect of 448-kHz capacitive resistive monopolar radiofrequency (CRMRF) in a porcine model. Materials and Methods This study investigated the effect of selective destruction of the subcutaneous fat layer in abdominal fat tissue using CRMRF. The effects of two types of CRMRF (capacitive electric transfer (CET) and resistive electric transfer (RET)) treatment were evaluated using regular digital photography in addition to thermal imaging evaluation, ultrasound measurement, hematological evaluation, and histologic analyses (H&E (hematoxylin and eosin), Oil red O, and immunohistochemistry staining). Results Preclinical evaluation was performed to obtain the data for comparison of the safety and efficacy of the subcutaneous fat reduction after applying CRMRF using CET and RET. After treatment, the thermal transmission was effective, and a 42-47℃ temperature change was observed in the fat layer while an approximately temperature of 42℃ was confirmed on the skin surface. Moreover, after the application of both types of CRMRF treatment, fibrotic septa were observed in the adipose tissue induced by heat at the treatment sites. TUNEL staining was also performed to confirm the process of apoptosis in the adipocytes. Conclusion These results suggest that both CET and RET for CRMRF treatment are safe and effective for subcutaneous fat reduction in a porcine model.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.123-129
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• 2011

The pig has been considered to serve as an appropriate model of human disease. Therefore, establishment of porcine embryonic stem cell lines is important. The purpose of the present study was to further work in this direction. We produced porcine parthenogenetic embryos, and separately aggregated two of each of two-cell ($2{\times}2$), four-cell ($2{\times}4$), and eight-cell ($2{\times}8$) embryos derived by parthenogenesis. After culture for 4 days, the developmental ability of the aggregates and total blastocyst cell numbers were evaluated. The percentage of blastocysts was significantly higher in both $2{\times}4$- and $2{\times}8$-aggregated embryos ($58.3{\pm}1.9%$ and $37.2{\pm}2.8%$, respectively) than in the control or $2{\times}2$-aggregated embryos ($23.6{\pm}1.1%$ and $12.5{\pm}2.4%$, respectively). Total blastocyst cell numbers were increased in the $2{\times}4$- and $2{\times}8$-aggregated embryos (by $44{\pm}3.0%$ and $45{\pm}3.3%$, respectively) compared with those of control or $2{\times}2$-aggregated embryos ($30.5{\pm}2.1%$ and $30.7{\pm}2.6%$, respectively; p<0.05). The levels of mRNA encoding Oct-4 were higher in both the $2{\times}4$- and $2{\times}8$-aggregated embryos than in the control. When blastocysts derived from $2{\times}4$- aggregated embryos or intact normal embryos were cultured on mouse embryonic fibroblast feeder cells to obtain porcine stem cells, blastocysts from aggregated embryos formed colonies that were better in shape compared with those derived from intact blastocysts. Together, the data show that aggregation of porcine embryos not only improves blastocyst quality but also serves as an efficient procedure by which porcine embryonic stem cells can become established.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.33 no.11
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• pp.1202-1208
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• 2009

An inverse finite element (FE) model parameter estimation algorithm can be used to characterize mechanical properties of biological tissues. Using this algorithm, we can consider the influence of material nonlinearity, contact mechanics, complex boundary conditions, and geometrical constraints in the modeling. In this study, biomechanical experiments on macro and micro samples are conducted and characterized with the developed algorithm. Macro scale experiments were performed to measure the force response of porcine livers against mechanical loadings using one-dimensional indentation device. The force response of the human liver cancer cells was also measured by the atomic force microscope (AFM). The mechanical behavior of porcine livers (macro) and human liver cancer cells (micro) were characterized with the algorithm via hyperelastic and linear viscoelastic models. The developed models are suitable for computing accurate reaction force on tools and deformation of biomechanical tissues.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.735-739
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• 2002

Large quantities of porcine hepatocyte aggregates with various degrees of aggregation (DA) could be obtained by controlling the suspension periods (0,9,24, and 48 h), and by entrapping the hepatocyte aggregates in model materials of encapsulation such as Ca-alginate and type-I collagen gels. The effects of DA on liver-specific functions of hepatocytes were evaluated in order to obtain optimum DA for the cell source of bioartificial liver (BAL) systems. Irregular rugged aggregates (size $75 \pm 28$ $\mu\textrm{m}$) farmed by 24 h of suspension culturing showed peak viability and hepatic functions such as ammonia removal and albumin secretion in the two types of entrapment systems, thus offering themselves as a stable cell source of a BAL system for hepatic functions and scale-up.