• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.187-191
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• 2007

This study was canted out to develop cell lines overexpressing human H-transferase (HT). One of the approaches to prevent hyperacute rejection in xenotransplantation might be the expression of human HT in porcine cells. In this study, we cloned human HT gene from HepG2 cells using RT-PCR to establish HT-overexpressing vector. The full-length cDNA of human HT was inserted into the 3' end of CMV promoter for construction of the overexpression vector pRc/CMV-hHT. Using ietPEI DNA transfection reagent, the vector was introduced into porcine ear skin fibroblasts from newborn piglets. Transfected cells were selected by treatment of $300{\mu}g/ml$ G418 for 12 days. After antibiotic selection, survived colonies with approximately 5mm in diameter were picked and analysed for transgene human HT by PCR. The colonies proven to be human HT transfectants were analysed by RT-PCR to determine their expressions or human HT. In all colonies tested, human HT mRNA was detected. This result demonstrates the establishment of porcine cell lines overexpressing human HT, and these cell lines may be used for the development of transgenic pigs for xenotransplantation.

• Korean Journal of Poultry Science
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• v.33 no.3
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• pp.181-188
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• 2006

Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.

• Journal of Radiation Protection and Research
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• v.36 no.2
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• pp.93-98
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• 2011

We induced the activation of melanocytes in the epidermis of C57BL/6 mice by ultraviolet-B (UV-B) irradiation, and observed the effect of an herbal preparation (HemoHIM, HH) on the formation, and decrease of UV-B-induced epidermal melanocytes. C57BL/6 mice were irradiated by UV-B $80\;mJ{\cdot}cm^{-2}$ ($0.5\;mW{\cdot}sec^{-1}$) daily for 7 days, and HH was intraperitoneally, orally or topically applied pre- or post-irradiation. For the estimation of change of epidermal melanocytes, light microscopic observation with dihydroxyphenylalanine (DOPA) stain was performed. Split epidermal sheets prepared from the ear of untreated mice exhibited 13~15 melanocytes${\cdot}mm^{-2}$, and one week after UV irradiation, the applied areas showed an increased number of strongly DOPA-positive melanocytes with stout dendrites. But intraperitoneal, oral or topical treatment with HH before each irradiation interrupted UV-B-induced pigmentation and resulted in a marked reduction in the number of epidermal melanocytes as compared to the number found in UV-B-irradiated, untreated control skin. The number and size of DOPA-positive epidermal melanocytes were also significantly decreased in intraperitoneally injected or topically applicated group after irradiation with HH at 3rd and 6th weeks after irradiation. The present study suggests the HH as inhibitor of UV-B-induced pigmentation, and depigmenting agent.

• Korean Journal of Poultry Science
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• v.32 no.2
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• pp.125-133
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• 2005

Morphometric changes in testicular Sertoli and Leydig cells from hatching to adulthood were studied using Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study was to understand the developmental phase of the Sertoli and Leydig cells with age. Testis of chickens was fixed by whole body perfusion using a fixative containing $2.5\%$ glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using 1 Um sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. The average volume of a testis of 1 week old Korean native chickens was determined as $0.148\;cm^3$ and the parameter increased linearly from 1 week to 21 weeks days $(28.86\;cm^3)$, and did not change from 21 weeks to 64 weeks. The volume density of the seminiferous tubules increased with age from $32.6\%$ at week 1 to $92.89\%$ at week 64. The volume density of the interstitium represents $67.4\%$ of the testicular parenchyma at week 1. This proportion progressively diminished during development to reach a value of $7.11\%$ at week 64. The volume density of the Leydig cells decreased almost linearly from 1 week $(4.9\%)$ to 14 weeks $(1.7\%)$ and remained unchanged thereafter. In contrast, the Sertoli cells occupied a volume density of $3.4\%$ at week 1, increased progressively up to 18 weeks of age $(10.79\%)$ and remained unchanged thereafter. The absolute volume of the Leydig and Sertoli cells per testis increased significantly from week 1 to week 21 but did not change significantly from week 24 to week 64. The number of Leydig cells per testis increased almost linearly from 1 week to 21 weeks, remained high and unchanged with advancing age. The number of Sertoli cells per testis increased gradually with age from 1 week to 14 weeks and remained unchanged thereafter.

• Korean Journal of Food Science and Technology
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• v.42 no.1
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• pp.109-114
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• 2010

The in vivo and in vitro effects of oriental herb extracts of Cassia obtusifolia, Taraxacum platycarpum and Ulmusmacrocarpa on anti-atopic allergic reaction were evaluated in this study. A mixture of these extracts exhibited more potent anti-allergic activities in human mast cells than those from individual extracts. The herbal mixture significantly inhibited the release of compound 48/80-induced $\beta$-hexosaminidase release in the human mast cell line, HMC-1. The mixture also suppressed the production of PMA and A23187-induced inflammatory cytokines in HMC-1 cells. To further investigate the in vivo effects of the herbal mixture, a Dermatophagoides farinae (DF)-induced atopic dermatitis mouse model was utilized. Oral administration of the herbal mixture significantly decreased the ear thickness and swelling in DF treated NC/Nga mice in a dose dependent manner. Furthermore, serum levels of IgE and interleukin-4 (IL-4) were significantly decreased, whereas interferon-gamma (IFN-$\gamma$) levels were increased in the mixture administrated groups when compared to the control. Taken together, our data indicate the possibility of using a mixture of the oriental herb extract to relieve symptoms of atopic dermatitis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.712-716
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• 2011

We investigated changes in the extraction yield and whitening activity of polysaccharides from Undaria pinnatifida sporophyll due to gamma irradiation. U. pinnatifida sporophyll was gamma-irradiated at doses of 10, 30, 50, 70, and 100 kGy, then extracted with hot water and precipitated with ethanol to extract polysaccharides. Crude polysaccharide yields increased with an increase in irradiation dosage, but tyrosinase inhibition activity did not change. Melanin synthesis did not significantly differ between B16BL6 cells treated with irradiated and non-irradiated samples. In conclusion, gamma irradiation increased the crude polysaccharide extraction yield but did not change the whitening activity of U. pinnatifida sporophyll. This implies that gamma irradiation can be used to increase yields in the cosmetic industry, thus increasing profits.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.8
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• pp.1090-1098
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• 2016

The anti-inflammatory effects of ethanol extract from Grateloupia crispata (GCEE) were investigated in lipopolysaccharide (LPS)-stimulated murine macrophages. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There was no cytotoxic effect on proliferation of macrophages treated with GCEE compared to the control. GCEE significantly inhibited production of pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis $factor-{\alpha}$, and $IL-1{\beta}$] as well as nitric oxide in LPS-stimulated RAW 264.7 cells. In addition, GCEE suppressed expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ in a dose-dependent manner. GCEE significantly reduced activation of mitogen-activated protein kinases. In the in vivo test, evaluation of anti-inflammatory activity of GCEE was performed using croton oil-induced ear edema in ICR mice. Oral administration of 10 mg/kg to 250 mg/kg of GCEE significantly reduced ear edema in a dose-dependent manner compared to croton oil-induced mice. Moreover, GCEE reduced ear thickness and the number of mast cells compared to croton oil-induced mice in the histological analysis. These data suggest that GCEE could be used as a potential source for anti-inflammatory agents.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.171-178
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• 2003

This study was conducted to investigate the effect of electric fusion methods on cell fusion rate and embryo development by somatic cell nuclear transfer in Korean Native Cattle. The KNC ear cell was cultured in vitro for confluence in serum starvation condition(DMEM＋0.05％ FBS) for cell confluence. The zona pellucida of IVM oocytes were partially dissection using micro pipette. Ear cells were transferred into an enucleated oocyte. The reconstructed embryos were electrically fused with Zimmermann Cell Fusion Medium(ZCFM). Nuclear transfer embryos were activated with a combination of 10${\mu}{\textrm}{m}$ calcium ionophore(5 min) and 2.0mM 6-DMAP(3 hr). The activated embryos were cultured in CR1 -aa medium contains 0.3％ BSA or 10％ FBS at 37$^{\circ}C$, 90％ $N_2$, and 5％ $CO_2$in incubator for 6 days. The fusion rates were 51.6％(chamber) and 68.9％(needle), respectively and there were significantly difference between the fusion method(P<0.05). But, lysis rates were not significantly different(10.7％, 11.5％), respectively. The cleavage rates were significantly different between the chamber method(73.2％) and needle method(80.3％), respectively(P<0.05). The rates of early embryos(2∼4cells) and blastocysts of chamber and needle methods were 54.1％, 61.1％ and 18.4％, 26.3％ respectively, and needle method was significantly higher than chamber method(P<0.05). But, morulae formation rate were not significantly differences between the chamber(6.7％) and needle(6.2) method(P <0.05). These result suggest that electric fusion of needle method was to be profitable for nuclear transfer embryo fusion rate, blastocyst formation rate and reduce of oocyte lysis.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.233-240
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• 2003

This study was conducted to examine the effects of oocyte maturation period, phytohemagglutinin-P (PHA-P) treatment and activation agent on the enucleation, fusion, activation or in vitro development of bovine nuclear transfer embryos. Bovine oocytes were enucleated at 16∼24 h of in vitro maturation (IVM). Adult ear skin cells treated or non-treated with PHA-P were transferred into enucleated oocytes. Reconstituted oocytes treated or non-treated with PHA-P were fused by a pulse of 1.5 kV/cm for 30 $\mu$sec. Fused oocytes were activated with a combination of calcium ionophore (A23187) and cycloheximide (CHXM) or dimethylaminopurine (DMAP), and cultured in vitro for 7∼9 days. Enucleation rate was significantly increased when oocytes were matured for 16∼18 h (70.2∼92.3%, P<0.05) compared to that of oocytes were matured for 20∼24 h (44.3∼53.4%). The location of metaphase-II plate was far off from the 1st polar body as maturation time was increased. PHA-P treatment of donor cells or reconstituted oocytes significantly improved fusion rate (P<0.05). Cleavage and blastocyst formation rates were significantly increased after activation with a combination of A23187 and DMAP (78.6% and 32.9%, respectively) compared to those of embryos activated with a combination of A23l87 and CHXM (48.5 and 15.2%, respectively). From the present result, it is suggested that high enucleation efficiency can obtained by using oocytes matured for 18 h. It also shows that PHA-P treatment can improve the fusion rate, and activation with a combination of A23187 and DMAP can enhance the embryo development.

• Korean Journal of Food Science and Technology
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• v.46 no.1
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• pp.87-93
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• 2014

We investigated the effects of an ethanol extract from Perilla frutescens sprouts (PFSE) as an antioxidant, and its effects on edema and inflammation in RAW 264.7 cells and HMC-1 cells. The antioxidant activities (DPPH and ABTS radical scavenging) of PFSE were similar to those of butylated hydroxytoluene (BHT) and (${\pm}$)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox). We also investigated the anti-inflammatory effects of PFSE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and HMC-1 cells stimulated with phorbol 12-myristate 13-acetate (PMA) with the calcium ionophore A23187. TNF-${\alpha}$ and IL-$1{\beta}$ production, which had been increased by treatment with LPS or PMA plus A23187, were significantly inhibited by PFSE in a dose-dependent manner. Furthermore, PFSE significantly reduced the xylene-induced ear edema and the carrageenan-induced paw edema of ICR mice in a dose-dependent manner. The effects of PFSE (200 mg/kg) in reducing ear and paw edema were similar to those of aspirin (50 mg/kg). These results suggest that PFSE can be potentially used as a medicine for treating oxidative stress, an edematous and inflammatory disease.