Current studies have revealed the capacity of mesenchymal stem cells (MSCs) in term of immunomodulatory properties, and this distinct potential is downgraded according to the disease duration of patients-derived MSCs. In order to enhance the immunomodulatory and anti-tumorigenic properties of the rheumatoid arthritis (RA) joints-derived MSCs, we aggregate synovial fluid-derived MSCs from RA joints (RA-hMSCs) into 3D-spheroids by the use of hanging drop culture method. Cells were isolated from synovial fluids of RA joints with longstanding active status over 13 years. For aggregation of RA-hMSCs into 3D-spheroids, cells were plated in hanging drops in 30 μL of advanced DMEM (ADMEM) containing 25,000-30,000 cells/ drop and cultured for 48 h. To analyze the comparative immunomodulatory effects of 3D-spheroid and 2D monolayer cultured RA-hMSCs and then cells were cultured in ADMEM supplemented with 20% of synovial fluids of RA patients for 48 h and were evaluated by qRT-PCR for their expression of mRNA levels of inflammatory and antiinflammatory markers. Cellular aggregation of RA-hMSCs was observed and cells were aggregate into a single sphere. Following treatment of RA patient’s synovial fluids into the RA-hMSCs, spheroids formed RA-hMSCs showed significantly (p < 0.05) higher expression of TNFα stimulated gene/protein 6 (TSG-6) than the monolayer cultured RAhMSCs. Therefore, the 3D-spheroid culture methods of RA-hMSCs were more effective than 2D monolayer cultures in suppressing inflammatory response treated with 20% of RA-synovial fluids by expression of TNFα (TSG-6) according to the immune response and enhanced secretion of inflammatory factors.

The estrogen-mediated effect of mesenchymal stem cells (MSCs) is a highly critical factor for the clinical application of MSCs. However, the present study is conducted on MSCs derived from adult donors, which have different physiological status with steroid hormonal changes. Therefore, we explores the important role of 17β-estradiol (E2) in MSCs derived from female and male newborn piglets (NF- and NM-pBMSCs), which are non-sexually matured donors with steroid hormones. The results revealed that in vitro treatment of MSCs with E2 improved cell proliferation, but the rates varied according to the gender of the newborn donors. Following in vitro treatment of newborn MSCs with E2, mRNA levels of Oct3/4 and Sox2 increased in both genders of MSCs and they may be correlated with both estrogen receptor α (ERα) and ERβ in NF-pBMSCs, but NM-pBMSCs were only correlated with ERα. Moreover, E2-treated NF-pBMSCs decreased in β-galactosidase activity but no influence on NM-pBMSCs. In E2-mediated differentiation capacity, E2 induced an increase in the osteogenic and chondrogenic abilities of both pBMSCs, but adipogenic ability may increased only in NF-pBMSCs. These results demonstrate that E2 could affect both genders of newborn donor-derived MSCs, but the regulatory role of E2 varies depending on gender-dependent characteristics even though the original newborn donors had not been affected by functional steroid hormones.

It is very difficult to get the information about semen quality analysis in transgenic pigs because of limited numbers and research facilities. Therefore, in the present study, we analyzed the semen quality of transgenic boars generated for xenotransplantation research. Briefly, the semen samples were collected from 5 homozygous α1,3-Galactosyltransferase knock-out (GalT-/-) transgenic boars and immediately transported to the laboratory. These semen samples were decupled with DPBS and conducted to analyze semen parameters by a computer-assisted semen analysis (CASA) system. The boar semen were examined all 12 parameters such as total motility (TM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and hyperactivated (HYP), etc. In results, among the 5 GalT-/- boars, three boars (#134, 144, and 170) showed normal range of semen parameters, but #199 and 171 boars showed abnormal ranges of semen parameters according to standard ranges of semen parameters. Unfortunately, #171 boar showed azoospermia symptom with rare sperm counts in the original semen. Conclusively, assessment of semen parameters by CASA system is useful to pre-screening of reproductively healthy boar prior to natural mating and artificial insemination for multiplication and breeding.

Abnormal epigenetic reprogramming of donor nuclei is supposed to be one of the factors that causes low development efficiency of mammalian somatic cell nuclear transfer (SCNT). Trichostatin A (TSA) is an inhibitor of histone acetylase, and so development of SCNT embryos could be increased by treatment with TSA. In the present study, we examined the effect of TSA on in vitro development of porcine embryos derived from NT (nuclear transfer) by investigating the status of histone acetylation in TSA-treated and control NT embryos and the expression of developmental related genes. In this study, we found that incubating　NT embryos with 40nM TSA for 24h after activation could improved the blastocyst formation rate from 13.7% to 32.5%. Thechange in histone acetylation level as a reslut of TSA treatment were validated using immunofluorescence and confocal microscopy. Immunofluorescence results indicated that the level of aetylation at histone 3 lysine 18 (AcH3K18) was increased at early embryo development stage after TSA treatment. furthermore, we compared the expression patterns of several genes (developmental related genes; Oct4, Sox2, Nanog, Cdx2, the imprinting genes; igf2r). TSA treatment improved the expression of development related genes such as Oct4, Cdx2, Nanog as well as the imprinted genes like igf2r. In conclusion, our results demonstrated that TSA treatment improves the in vitro development of porcine NT embryos, increased the global histone acetylation (AcH3K18) and enhances the expression of some developmentally important genes (Oct4, Cdx2, Nanog) at blastocyst stages.

In reality, it is a green revolution of the entire agricultural matrix in Korea that integrated pest control plays an important role in the possible breakthrough in rice self-sufficiency. In paddy agroecosystem as man-modified environment, rice is newly established every year by transplantation under diverse water regimes which affect a microclimate. Standing water benefits rice by regulating the microclimate, but it favors the multiplication of certain pets through the amelioration of the microclimate. Further, the introduction of high yielding varieties with the changing of cultural practices results in changing occurrence pattern of certain pests. In general, japonica type varieties lack genes resistant to most of the important pests and insect-borne virus diseases, whereas indica type possesses more genes conferring varietal resistance. Thus, this differences among indica type, form the background of different approaches to pest management. The changes in rice cultivation such as double cropping, growing high-yielding varieties requiring heavy fertilization, earlier transplanting, intensvie-spacing transplanting, and intensive pesticide use as a consequence of the adoption of improves rice production technology, have intensified the pest problems rather than reduced them. The cultivation of resistant varieties are highly effective to the pest, their long term stability is threathened because of the development of new biotypes which can detroy these varieties. So far, three biotypes of N. lugens are reported in Korea. Since each resistant variety is expected to maintain several years the sequential release of another new variety with a different gene at intervals is practised as a gene rotation program. Another approach, breeding multilines that have more than two genes for resistance in a variety are successfully demonstrated. The average annual rice losses during the last 15 years of 1977-’91 are 9.3% due to insect pests without chemical control undertaken, wehreas there is a average 2.4% despite farmers’insecticide application at the same period. In other words, the average annual losses are prvented by 6.9% when chemical control is properly employed. However, the continuous use of a same group of insecticides is followed by the development of pest resistance. Resistant development of C. suppressalis, L. striatellus and N. cincticeps is observed to organophosphorous insecticides by the mid-1960s, and to carbamates by the early 1970s in various parts of the country. Thus, it is apparent that a scheduled chemical control for rice production systems becomes uneconomical and that a reduction in energy input without impairing the rice yield, is necessarily improved through the implementation of integrated pest management systems. Nationwide pest forecasting system conducted by the government organization is a unique network of investigation for purpose of making pest control timely in terms of economic thresholds. A wise plant protection is expected to establish pest management systems in appropriate integration of resistant varieties, biological agents, cultural practices and other measures in harmony with minimizing use of chemical applications as a last weapon relying on economic thresholds.

주차장 바닥재로 많이 사용되는 A사 에폭시 코팅제와 B사 바닥코팅제(A, B type)에 대하여 GC/MS(Gaschromatography /Mass spectrometer)를 이용하여 제품별로 함유하고 있는 휘발성유기화합물(VOCs)의 종류와 함량을 측정해보고, 휘발성유기화합물(VOCs)중 작업자나 입주자들에게 유해한 폼알데하이드(Formaldehyde), 톨루엔(Toluene), 크실렌(Xylene)을 외부 환경에 따른 오차를 줄이고자 가장 안 좋은 (밀폐) 환경을 가정하고 가스검지관을 이용하여 가스배출량을 측정해 보았다. 그 결과 A사 에폭시코팅제가 제품 자체에도 휘발성유기화합물(VOCs)을 가장 많은 양 함유하고 있고, A사 에폭시코팅제에 비하여 B사 바닥코팅제 A type은 약 79%, B type은 약 96% 이상의 적은 양을 나타냈다. 또한, 밀폐된 환경 조건에서 1시간과 8시간 경과 후 가스검지관을 이용하여 폼알데하이드(Formaldehyde), 톨루엔(Toluene), 크실렌(Xylene)에 대한 가스배출량을 측정하고 TWA값으로 환산한 결과에서도 A사 에폭시코팅제가 가장 높게 측정되었다. A사 에폭시코팅제보다 B사 바닥코팅제 A type은 약 42.3%이상의 적은 측정값을 나타냈으며, B type의 경우 밀폐된 환경 조건에서도 시간가중평균노출기준(TWA)을 모두 만족하였다.

One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-humanprimate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were 127 ± 18.9. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as α-1,3-galactosyltransferase knock-out /hCD46 for xenotransplantation.