Diabetes mellitus, the most common metabolic disorder, is divided into two types: type 1 and type 2. The essential treatment of type 1 diabetes, caused by immune-mediated destruction of β-cells, is transplantation of the pancreas; however, this treatment is limited by issues such as the lack of donors for islet transplantation and immune rejection. As an alternative approach, stem cell therapy has been used as a new tool. The present study revealed that bone marrowderived mesenchymal stromal cells (BM-MSCs) could be transdifferentiated into pancreatic cells by the insertion of a key gene for embryonic development of the pancreas, the pancreatic and duodenal homeobox factor 1 (PDX1). To avoid immune rejection associated with xenotransplantation and to develop a new cell-based treatment, BM-MSCs from α-1,3-galactosyltransferase knockout (GalT KO) pigs were used as the source of the cells. Transfection of the EGFP-hPDX1 gene into GalT KO pig-derived BM-MSCs was performed by electroporation. Cells were evaluated for hPDX1 expression by immunofluorescence and RT-PCR. Transdifferentiation into pancreatic cells was confirmed by morphological transformation, immunofluorescence, and endogenous pPDX1 gene expression. At 3∼4 weeks after transduction, cell morphology changed from spindle-like shape to round shape, similar to that observed in cuboidal epithelium expressing EGFP. Results of RT-PCR confirmed the expression of both exogenous hPDX1 and endogenous pPDX1. Therefore, GalT KO pig-derived BM-MSCs transdifferentiated into pancreatic cells by transfection of hPDX1. The present results are indicative of the therapeutic potential of PDX1-expressing GalT KO pig-derived BM-MSCs in β-cell replacement. This potential needs to be explored further by using in vivo studies to confirm these findings.
Pluripotency of human embryonic stem cell (hESC) is one of the most valuable ability of hESCs for applying cell therapy field, but also showing side effect, for example teratoma formation. When transplant multipotent stem cell, such as mesnchymal stem cell (MSC) which retains similar differentiation ability, they do not form teratoma in vivo, but there exist limitation of cellular source supply. Accordingly, differentiation of hESC into MSC will be promising cellular source with strong points of both hESC and MSC line. In this study, we described the derivation of MSC like cell population from feeder free cultured hESC (hESC- MSC) using direct differentiation system. Cells population, hESC-MSC and bone marrow derived MSC (BM-MSC) retained similar characteristics in vitro, such as morphology, MSC specific marker expression and differentiation capacity. At the point of differentiation of both cell populations, differentiation rate was slower in hESC-MSC than BM-MSC. As these reason, to verify differentially expressed molecular condition of both cell population which bring out different differentiation rate, we compare the molecular condition of hESC-MSC and BM-MSC using 2-D proteomic analysis tool. In the proteomic analysis, we identified 49 differentially expressed proteins in hESC-MSC and BM-MSC, and they involved in different biological process such as positive regulation of molecular function, biological process, cellular metabolic process, nitrogen compound metabolic process, macromolecule metabolic process, metabolic process, molecular function, and positive regulation of molecular function and regulation of ubiquitin protein ligase activity during mitotic cell cycle, cellular response to stress, and RNA localization. As the related function of differentially expressed proteins, we sought to these proteins were key regulators which contribute to their differentiation rate, developmental process and cell proliferation. Our results suggest that the expressions of these proteins between the hESC-MSC and BM-MSC, could give to us further evidence for hESC differentiation into the mesenchymal stem cell is associated with a differentiation factor. As the initial step to understand fundamental difference of hESC-MSC and BM-MSC, we sought to investigate different protein expression profile. And the grafting of hESC differentiation into MSC and their comparative proteomic analysis will be positively contribute to cell therapy without cellular source limitation, also with exact background of their molecular condition.
Aujeszky’s disease (AD), also called pseudorabies, is an infectious viral disease, caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. AD affects many countries and regions in the world, causing important economic losses, mainly due to international trade restrictions. In this study, to determine the characteristics of the Aujeszky’s disease virus (ADV), NYJ strain, which was isolated from the serum of an infected pig in 1987, we investigated the nucleotide sequence and expression of the glycoproteins gB, gC, and gD using the bBpGOZA system. We found that the glycoproteins gB, gC, and gD of NYJ consisted of 2751 bp, 1443 bp, and 1203 bp, respectively. Comparison of the NYJ with the other strains revealed nucleotide sequence identity ranging from 91.tito 99.0%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The NYJ strain was formed a distinct branch with high bootstrap support. The expression of glycoprotein gD in insect cells was characterized by SDS-PAGE and Western blotting with an anti-ADV polyclonal antibody. Glycoprotein gD of approximately 45 kDa was detected. The results of this study have implications for both the taxonomy of ADV and vaccine development.
The porcine reproductive and respiratory syndrome virus (PRRSV) has six structural proteins which encoded by ORFs 2 to 7 are designated as GP2, 3, 4, 5, M and N, repectively. In this study, we determined the expression of each protein using novel transfer vector, pBmKSK4 which has the polyhedrin promoter of BmNPV and 6xHis tag. The recombinant transfer vector was co-transfected into Bm5 cells along with bBpGOZA DNA. Recombinant virus was purified by plaque assay and amplified in Bm5 cells. Expression of each protein was identified by SDS-PAGE and Western blot analysis using anti-6xHis monoclonal antibody. The expression levels of the structural proteins in Bm5 cells were stronger than the expression system using pBacPAK9 transfer vector in Sf21 cells. As expected, GP5 was expressed at low levels from its structural properties and its toxicity for cells. In addition, each recombinant protein was purified using Ni-NTA spin columns. The ability to produce each protein in the baculovirus system indicates that these could be major candidates for the development of a vaccine against PRRSV.
본 고에서는 BM특허 및 의약특허의 미완성발
명 및 명세서 기재불비관련 최근 주요 판례에서 판
단기준을 재해석하고 미완성 발명과 명세서 기재
불비의 상호관계를 정립함으로써 심사 또는 심판
처리에서의 정확성 및 공정성을 전보다 한층 더 도
모하고자 한다.
BM특허 및 의약특허의 경우 구체성이 없고 현
재의 기술수준으로는 실시가 불가능한 경우에도
미완성발명으로 취급하여 특허법제29조제1항본
문을 적용하여 명세서 기재불비와 구별할 필요가
있으며, 그 이외의 사항에 대해서는 특허법제42조
제3항의 명세서 기재불비로 거절결정하는 것이 심
사 심판실무에서 바람직 할 수 있다.
종래에는 산업에 해당되지 않는다고 생각되어
온 의료관련 행위가 바이오 기술의 발전에 따라 제
한 없이“산업화”경향을 나타내고 있으며, 오히려
의료업 자체가 신규산업의 창출과 산업 발전에 불
가결한 것으로 여겨지고 있다. BM 기술의 인터넷
산업도 상기와 거의 같다.
결론적으로, SW특허 및 BM특허 등 모든 기술
분야에 예외 없이 특허보호를 인정하려는 추세를
고려하여 볼 때, 법적 해결이 아닌 법해석에 의해
거절이유를 제시한다는 것은 적절치 않다. 즉, 진정
한 산업발전을 위해서는 특허법의 취급을 해석론
의 틀에서 논쟁을 벌일 것이 아니라 입법적으로 해
결해야 한다고 본다.
A new display device is required, which has concepts of flatness and slimness. FED can be one of the solutions. When we use flat panel, we can save the raw material and reduce the production time by eliminating the printing process, drying process, and wa
A new display device development using CRT and CDT technology is required, which has concepts of flatness and slimness. Screen printing technology can be one of the solutions. In this case, good panel flatness is the precondition. Therefor, we did process capability analysis of panel flatness and regression analysis between panel flatness and BM(black matrix) position by experiments.
In this study, β-1,3/1,6-glucan, lactic acid bacteria, and β-1,3/1,6-glucan+lactic acid bacteria were tested for 10 weeks using an immunodeficient animal model infected with LP-BM5 murine AIDS virus On the immune activity. Cytokines production, plasma immunoglobulin concentration, T cell and B cell proliferation were measured. As a result, the T cell proliferative capacity which was weakened by immunization with LP-BM5 murine AIDS virus increased significantly T cell proliferative capacity compared with the red ginseng control group. B cell proliferative capacity was significantly higher than the infected control group. Increased B cell proliferation was reduced. In the cytokine production, IL-2, IL-12 and IL-15 in the Th1-type cytokine increased the secretion of IL-2, IL-12 and IL-15 compared to the infected control. The proliferative capacity of the treated group was higher than that of the mixed treatment group. TNF-α was significantly decreased compared with the infected control group. The IL-4, IL-6 and IL-10 levels were significantly inhibited in the infected control group and the Th1/Th2 type cytokine expression was regulated by immunohistochemistry. IgE, IgA, and IgG levels were significantly lower in the immunoglobulin secretion assay than in the control. As a result, the immunomodulatory effect of β-1,3/1,6-glucan+lactic acid bacteria was confirmed by mixing with LP-BM5 murine AIDS virus-infected immunodeficient animal model.
Background : barley (Hordeum vulgare L.), is a member of the grass family Poaceae (Gramineae) consider the fourth most important cereal crop and primarily used to feed the animals. The second most common use of barely is for malt, which is used mostly in beer, but also in hard liquor, malted milk and flavoring in a variety of foods. As immense economic importance a fast and high germination is required in order to achieve its desired production. In regard to this the present study was performed to evaluate the effect of beneficial microbes (BM), on barely seed germination when mixed with in the synthetic germination medium. Methods and Results : Barely seeds were surface sterilized using 0.01% HgCl2 solution and transfer for germination in the medium supplemented with 200 and 500 X diluted BM separately. Effectiveness of the BM were determined by calculating the germination percentage (GP), germination index (GI), mean germination time (MGT), germination time 50% (T50) and compared with control seeds germinating in absence of BM. Results obtained from the study suggest that BM at 200 X dilution is much efficient to speed up the germination process (66.7 ± 2.3 %) in contrast to 500X diluted BM (61.4 ± 3.6 %). Results of the other germination parameters (GI, MGT and T50) supported the 200X BM effective for seed germination as compare to 500X and control. Conclusion : Our results signify that BM has potential to enhance the germination efficacy of barley seeds and thus could be used to improve the germination.
Recycling of food wastes was tried based on fermenting and composting food wastes using a microbial consortium. Manufactured compost (using 11.3% food waste) turned out to be effective in increasing soil fertility and crop growth (radish; Raphanus sativus). More specifically, the treatment of the composted food wastes enabled a stimulated growth of radish leaves by 80% and an increased uptake of δ15NAIR by 250% compared with a commercial organic compost. Moreover, the compost derived from the wastes appeared to allow a sustainable management of nitrogen fertilizer compared with the chemical fertilizer, minimizing nitrogen pollution. The microbial community analysis showed significant difference in the microbial community pattern in soil treated with the composted food wastes relative to soil treated with a commercial organic fertilizer or a chemical fertilizer. The results may indicate that the wastes processed by the consortium could result in an efficient recycling of the nuisance materials such as food wastes and other organic solid wastes.
본 연구에서는 선박에서 발생하는 오․폐수의 처리를 위하여 SBR공정에 유효미생물을 주입하는 변법을 이용하여 Lab scale 실험을 수행하였다. 유해물질 유입에 따른 생물학적 처리 장치의 효율 저하 문제를 해결하기 위하여 SBR공정에 유효미생물을 주입하는 변법은 크루 즈선이라는 특수 환경과의 접목성과 생물학적 처리 시 야기될 수 있는 문제를 대비하기 위한 대안으로 선박환경에 매우 적합한 공정으로 평가 되었다. 슬러지 관찰 결과 기존의 활성슬러지에 유효미생물의 주입함으로써 슬러지의 안정성을 확보할 수 있었으며, 슬러지의 EPS 함량도 40% 이상 높아진 것을 확인할 수 있었다. 또한 슬러지의 미생물 분석 결과 유효미생물 주입으로 인해 수처리에 유리한 미생물종이 다수 출현 하여 휘발성 유기화합물과 같은 유기 유해물질이 생분해되어 안전한 물질로 전환되는 것을 확인할 수 있었으며. 중금속과 같은 무기 유해물질 도 중금속의 종류와 유입농도에 영향을 받지 않고 70% 이상의 안정적인 처리 효율도 확보할 수 있는 것으로 확인되었다.
본 연구에서는 유입수의 변동이 심하고 전문가가 부재한 환경인 선박에서 발생하는 오수의 효과적인 처리를 위하여 RCM공법을 선박오수처리장치에 적용하는 실험실 규모의 실험을 수행하였다. 질소·인의 고도처리 효율과 선박이라는 특수환경과의 접목성을 검토한 결과 RCM공정에 유효미생물을 주입하는 방법은 선박환경에 적합한 것으로 평가되었다. 또한 RCM공정은 활성슬러지 공정에서 배출되는 슬러지 는 배출시키지 않고 슬러지액화분해조(SDC)에서 재분해하여 순환함으로써, 최근 해양투기가 금지됨으로 인해 문제가 되고 있는 슬러지의 발 생량을 최소한으로 하여 친환경적인 수처리가 가능하다. 복합미생물제제 주입 후 미생물 관찰결과 고도처리에 유리한 미생물종의 출현을 확 인하였으며 이들의 상호기작으로 질소·인의 처리에 도움을 주어 처리효율이 높은것이라 판단된다. 유기물 제거효율 실험결과 BOD5, CODcr T-N, T-P의 처리효율이 각각 96, 97, 78, 81.68 %로 나타나 Membrane이나 Filter없이도 강화되어가는 해양오염기준을 충족시킬 수 있는 공 정으로 판단된다.
본 연구에서는 크루즈선에서 발생하는 오 폐수의 고도처리를 위하여 SBR공정에 유효미생물을 주입하는 변법을 이용하여 실험실 규모의 실험을 수행하였다. 질소 인의 고도 처리 효율과 크루즈선이라는 특수 환경과의 접목성을 검토한 결과 SBR공정에 유효미생물을 주입하는 변법은 선박환경에 매우 적합한 것으로 평가되었다. 또한 기존의 활성슬러지에 유효미생물의 주입함으로써 슬러지 팽화 등의 문제를 해결할 수 있어 슬러지의 안정성을 확보할 수 있었으며, 미생물 관찰 결과 고도처리에 유리한 미생물종의 출현으로 질소 인의 처리 효율이 높아지는 것을 확인할 수 있었다. 이로 인해 총질소와 인의 제거 효율은 74%, 75%로 나타났으며 이러한 결과로 미루어 강화되어가는 해양오염기준을 충만족시킬 수 있을 공정으로 판단된다.
본 연구는 크루즈선에서 발생하는 오 폐수의 생물학적 처리시 야기될 수 있는 악취 문제를 해결하기 위하여 SBR공정에 유효미생물을 주입하여 Lab scale 실험을 수행하였다. 악취물질의 저감 정도와 크루즈선이라는 특수 환경과의 접목성을 검토한 결과 유효미생물의 주입은 선박환경에 매우 적합한 공정으로 평가되었다. 기존의 활성슬러지에 유효미생물의 주입함으로써 슬러지 내 미생물의 다양성과 높은 개체수를 확보할 수 있었으며, 미생물 간의 우점종도 악취처리에 유리한 미생물종으로 전환되는 것을 확인할 수 있었다. 또한 이로 인하여 악취물질의 악취 강도도 20배 이상 낮출 수 있는 것으로 확인되었다.