• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.77-77
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• 2003

Chimeric animals are referred to as an organism composed of tissues derived from more than one species. In order to examine if a pluripotency of embryonic stem cells can cross the limitation of a species, we tried to establish human-mouse chimeric animals. Human embryonic stem cells were genetically modified to express eGFP using eukaryonic expression vector pcDNA 3.1 (In Vitrogene) for an easy identification. After selection with neomycin, approximately 15 cells were implanted into mouse blastocoele cavity. Ten chimeric blastocysts were transferred to one of the uterine horn of 2.5 days pesudopregnent ICR female. Out of 272 blastocysts transferred to pseudopregnant recipients 20 live newborn were obtained after 20 days. When newborn were obtained, pups were quickly removed immersed into 4% PFA. By histological examination using fluorescent microscope, green fluorescence was observed from the liver, heart, and spleen in newborn mice. Three weeks after born, presence of eGFP sequence within mouse genome (tail and kidney) was reconfirmed by PCR. eGFP sequence was amplified from the progenies of the animal suggesting a genetic transmission of the transgene. These chimeric mice having human cells at the beginning of development, are expected to recognize human cells as “self”, therefore, human cells or tissues will be able to escape the immunological surveillance of the host if grafted into the animal. These animals will serve as a good model system for studying the graft rejection in tissue transplantation and the potential of the cells to work well in many human disease.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.88-88
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• 2003

본 연구는 배란후 48시간째 회수된 난소로부터 황체세포를 체외배양 후 progesterone 분비에 대한 LH, IGF-1 및 TGF$\beta$첨가효과에 대해 검토하였다. 축산기술연구소에서 사육중인 돼지(체중 145$\pm$kg) 12두로부터 발정을 유도시켜 배란후 약 48시간째 도축하여 난소를 회수하였다. 회수된 난소로부터 황체를 분리하여 세절한 후 0.25% collagenase용액(0.025mg DNase, 50mM EDTA, 50mM Dithiothreitol)으로 37$^{\circ}C$의 진탕수조에서 30분간 배양하여 황체세포를 분리 회수하였다. 회수된 황체세포는 D-MEM 용액(GIBCO, 10% FCS와 antibiotics 첨가)으로 2회 세정하여 1$\times$$10^{6}$live cell/$m\ell$이 되도록 희석한 후 24 well culture plate(Coming, New Tork 14831)에 분주하여 $CO_2$ 배양기($CO_2$: 5%)에서 12시간 간격으로 배양액을 회수하였으며 48시간까지 배양하였다. 배양액 1$m\ell$당 LH, IGF-I 및 TGF$\beta$1을 단독 혹은 공배양하여 회수된 배양액내의 progesterone 농도를 RIA법으로 분석하였다. 돼지황체세포는 배양후 24시간째에 높은 농도의 progesterone이 측정되었으며 LH를 첨가한 대조구에 비해 유의적으로 높은 progesterone이 측정되었다. 또한 IGF-1 과 TGF$\beta$1을 첨가한 군에서도 높은 농도의 progesterone이 측정되었다 그러나 TGF$\beta$ 혹은 IGF-I 단독으로는 대조구의 결과와 큰 차이가 없었다 따라서 돼지 황체세포에서의 progesterone 분비는 TGF$\beta$ 및 IGF-I 그리고 LH의 황체세포의 progesterone 분비기능을 촉진하는 것으로 나타났다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권10호
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• pp.1557-1566
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• 2007

Ten male crossbred (Thai $Native{\times}Anglo$ Nubian) goats with average live weight of $27{\pm}2$ kg were randomly assigned according to a $5{\times}5$ replicated Latin square design to receive five diets, $T_1$ = concentrate with 0% cassava chip (CC), $T_2$ = 25% CC, $T_3$ = 50% CC, $T_4$ = 75% CC and $T_5$ = 100% CC. Fresh elephant grass (FEG) was offered ad libitum as the roughage. A metabolism trial lasted for 21 days during which liveweight changes and feed intakes were measured. Based on this experiment, there were no significant differences (p>0.05) among treatment groups regarding DM intake and digestion coefficients of nutrients (DM, OM, NDF and ADF), except for $T_5$ (100% CC) which was lowest (p<0.05) in digestion coefficient of CP than $T_1$ and $T_3$. Rumen parameters (ruminal pH, $NH_3$-N and volatile fatty acids), blood urea nitrogen, blood glucose and packed cell volume were similar among treatments. Moreover, rumen microorganism populations were not affected (p>0.05) by cassava inclusion. The amount of N absorption and retention were similar among treatments, except for $T_5$ which tended to be slightly lower. Based on this experiment, it could be concluded that the optimal level of cassava inclusion to replace corn in goat diets was in the range of 25-75% of CC when fed with FEG and it was a good approach in exploiting the use of local feed resources for goat production.

• 한국동물생명공학회지
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• 제35권2호
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• pp.119-141
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• 2020

The Dromedary camel (Camelus dromedaries) is an important species because of its ability to produce good quality meat, milk, and fibers under harsh environmental conditions. Camels are also crucial for transportation, racing, and as draft animals in agriculture. Therefore, dromedary camels play a critical role in the economy for millions of people living in the arid part of the world. The inherent capability of camels to produce meat and milk is highly correlated with their reproductive performance. Compared with other domestic species, the reproductive efficiency in camelids is low. Although recent reproductive technologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) have been successfully applied to camelids and the birth of live offspring following these technologies has been reported; in vitro embryo production (IVP) has lagged in this species. The development of the IVP system for dromedary camels may be a useful tool for the genetic improvement of this species. IVP in farm animals includes three main steps; in vitro maturation (IVM) of an oocyte, IVF of a matured oocyte, and in vitro culture (IVC) of fertilized oocyte up to the blastocyst stage. This review aims to summarize various factors that influence oocyte quality, IVM, and in vitro embryo development in dromedary camel.

• Reproductive and Developmental Biology
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• 제32권2호
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• pp.123-126
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• 2008

The objective of this study was to determine the potential hazardous effects of sorting process by flowcytometry on the quality of boar spermatozoa by flowcytometer. Freshly collected boar semen was diluted and divided into two groups; control none sorted and sorted. Sperms in sorted group were processed with flowcytometer for cell sorting with $100\;{\mu}M$ nozzle under the 20 psi pressure. Measurements on each parameter were made at two time points, 0hr (right after sorting) and 24 hr post sorting. Although there was a tendency of lower viability in sorted group than none sorted control group, the percentage of live cells in control ($75.83{\pm}6.92\;&\;59.53{\pm}10.34$) was not significantly different from sorted ($59.70{\pm}7.37\;&\;43.97{\pm}3.76$) at both 0 and 24 hr post sorting. However, sorted sperm showed significantly lower mitochondrial function compared to the control at both 0 h ($79.37{\pm}3.22\;vs.\;63.50{\pm}10.05$) and 24 hr ($67.27{\pm}3.22$ vs. $46.97{\pm}5.37$) time points (p<0.007). Sperm DNA fragmentation rate was significantly lower in control ($22.0{\pm}7.04$) than that of sorted ($32.27{\pm}7.49$) at 24 hr time point (p<0.0002). Taken together, these data suggested thatsorting process by flowcytometer may have influenced sperm motility rather than viability. Also high speed sperm sorting by flowcytometer has significant effects on DNA fragmentation on elapsed time after sorting.

• Journal of Microbiology and Biotechnology
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• 제26권3호
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• pp.469-476
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• 2016

This study was undertaken to evaluate the immunomodulatory effect of dead nano-sized Lactobacillus plantarum (nLp) in RAW 264.7 cells and murine primary splenocytes. nLp is a dead, shrunken, processed form of L. plantarum nF1 isolated from kimchi (a traditional Korean fermented cabbage) and is less than 1 μm in size. It was found that nLp treatment stimulated nitric oxide (NO) production more in RAW 264.7 macrophages than pure live L. plantarum (pLp), and that the stimulatory properties were probably largely derived from its cell wall. In addition, nLp induced murine splenocyte proliferation more so than pLp; in particular, a high dose of nLp (1.0 × 10¹¹ CFU/ml) stimulated proliferation as much as lipopolysaccharide at 2 μg/ml. Moreover, according to our cytokine profile results in splenocytes, nLp treatment promoted Th1 (TNF-α, IL-12 p70) responses rather than Th2 (IL-4, IL-5) responses and also increased Th17 (IL-6, IL-17A) responses. Thus, nLp stimulated NO release in RAW 264.7 cells and induced splenocyte proliferation more so than pLp and stimulated Th1 and Th17 cytokine production. These findings suggested that dead nLp has potential use as a functional food ingredient to improve the immune response, and especially as a means of inducing Th1/Th17 immune responses.

• Journal of Microbiology and Biotechnology
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• 제26권7호
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• pp.1198-1205
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• 2016

Lactic acid bacteria (LAB) have beneficial effects on intestinal health and skin diseases. Lipoteichoic acid (LTA), a cell wall component of gram-positive bacteria, is known to induce the production of several cytokines such as TNF-α, IL-1β, and IL-8 and affect the intestinal microflora, anti-aging, sepsis, and cholesterol level. In this study, Weissella cibaria was isolated from Indian dairy products, and we examined its immune-enhancing effects. Live and heat-killed W. cibaria did not induce the secretion of immune-related cytokines, whereas LTA isolated from W. cibaria (cLTA) significantly increased the secretion of TNF-α, IL-1β, and IL-6 in a dose-dependent manner. cLTA increased the phosphorylation of nuclear factor kappa-light-chain-enhancer of activated B cells, p38 mitogen-activated protein kinases, and c-Jun N-terminal kinases in THP-1 cells. The secretion of TNF-α and IL-6 was also increased in the cLTA-treated mouse splenocytes. These results suggest that cLTA, but not W. cibaria whole cells, has immune-boosting potential and can be used to treat immunosuppression diseases.

• 대한수의학회지
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• 제43권4호
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• pp.689-696
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• 2003

Canine sarcoptic mite (Sarcoptes scabiei var. canis) burrow usually in the stratum corneum of the skin of dogs and rabbits. Antigens from the burrowing mites induce cutaneous inflammatory reaction and humoral and cell-mediated immune response in the host. The effect of immunization induced by somatic antigens of house dust mite (Dermatophagoides spp.) has been evaluated to control the canine sarcoptic mite in this experiment. Twelve common antigens (187, 142, 126, 120, 109, 92, 80, 68, 51, 30, 25, 17 kDa) were found using SDS-PAGE with silver staining and Western blot between canine sarcoptic mite and house dust mite. In order to evaluate the immunologic effect of these common antigens 10 New Zealand white rabbits were divided as 4 groups such as negative control (group I), positive challenged control (group II), vaccinated (group III), and vaccinated-challenged (group IV) groups. Group II was artificially infested with about 1,000 canine sarcoptic mites and group III and IV were immunized with somatic antigens of house dust mite. In addition group IV was artificially infested with about 1,000 canine sarcoptic mites and group II, IV were treated with ivermectin. At the 8 weeks of the vaccination with common antigen, the antibody titers of all groups of II, III and IV had been increased. Both infestation score and live canine sarcoptic mite counts of group IV were lower than group III. Infestation score of group II become 0 by 2 weeks and group IV by 4 weeks after infestation. These results suggest that house dust mite, which is easy to culture in vitro, can be a vaccine candidate for protection of canine sarcoptic mite infestation.

• International Journal of Oral Biology
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• 제35권2호
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• pp.43-49
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• 2010

Enterococcus faecalis, a gram-positive bacterium, has been implicated in endodontic infections, particularly in chronic apical periodontitis. Proinflammatory cytokines, including tumor necrosis factor-$\alpha$ (TNF-$\alpha$), are involved in the pathogenesis of these apical lesions. E. faecalis has been reported to stimulate macrophages to produce TNF-$\alpha$. The present study investigated the mechanisms involved in TNF-$\alpha$ production by a murine macrophage cell line, RAW 264.7 in response to exposure to E. faecalis. Both live and heat-killed E. faecalis induced high levels of gene expression and protein release of TNF-$\alpha$. Treatment of RAW 264.7 cells with cytochalasin D, an inhibitor of endocytosis, prevented the mRNA up-regulation of TNF-$\alpha$ by E. faecalis. In addition, antioxidant treatment reduced TNF-$\alpha$ production to baseline levels. Inhibition of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinase also significantly attenuated E. faecalis-induced TNF-$\alpha$ expression by RAW 264.7 cells. Furthermore, activation of NF-${\kappa}B$ and AP-1 in RAW 264.7 cells was also stimulated by E. faecalis. These results suggest that the phagocytic uptake of bacteria is necessary for the induction of TNF-$\alpha$ in E. faecalis-stimulated macrophages, and that the underlying intracellular signaling pathways involve reactive oxygen species, ERK, p38 MAP kinase, NF-${\kappa}B$, and AP-1.

• 한국환경보건학회:학술대회논문집
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• 한국환경보건학회 2002년도 춘계 국제 학술대회
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• pp.49-51
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• 2002

To identify and evaluate the dichlorobenzidine(DCB)-DNA adducts in liver cell and bladder epithelial cells by $^{32}$ P-postlabeling and GC/MS-SIM, we orally exposed the dichlorobenzidine (20mg/kh body wt.,/day)to male sprague-dawley rats for 14 days. Two kinds of DCB-DNA adduct were found at the same site of thin layer chromatogram of $^{32}$ P-postlabeling method in liver cells and bladder epithelial cells. In liver cells, relative adduct labeling(RAL) $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 34.1$\pm$3.71 and 69.9$\pm$5.02, that of adduct A2 were 74.1$\pm$10.1 and 105.1$\pm$10.1 on 10 and 14 days after treatment, respectively. And in bladder epithelia cells, RAL $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 5.92$\pm$1.60 and 15.9$\pm$1.31, that of adduct A2 were 9,81$\pm$2.81 and 22.8$\pm$1.79 on 10 and 14 days after treatment, respectively. DCB metabolites formed DNA adducts were monoacetyl-dichlorobenzidine(acDCB) and diacety1-dichlorobenzidine(di-acDCB), which was identify by gas chromatography/mass spectrometry-scan ionization mode(GC/MS-SIM), along with hydrolysis, extraction and TFA(trifluoroacetyl anhyride) derivatization with DCB-DNA adducts isolated from live cells and bladder epithelial cells. The base peak of acDCB were 252 and 294 m/z, and that of di-acDCB were 252, 294 and 336 m/z. In conclusion, the exposed DCB formed two kinds of DCB-DNA adduct, the proximate materials of that were acDCB and di-acDCB in liver and bladder epithlial cells. And the above GC/MS-SIM method was found the DCB-DNA adducts could be monitoring by gas chromatography.