민법 제812조 제1항은 “혼인은 「가족관계의 등록 등에 관한 법률」에 정한 바에 의하여 신고함으로써 그 효력이 생긴다.”고 규정하여 법률 혼주의를 채택하고 있다. 이에 따라 혼인에 관한 규정은 혼인신고를 한 법률상 부부에게 적용이 되며 혼인신고를 마치지 않은 사실혼은 원칙적으로 인정되지 않는다. 다만 사실혼을 보호할 필요성이 있는 경우 혼인 신고를 전제로 한 효과를 제외한 나머지 법률혼의 효과를 인정한다. 판례에 따르면 중혼적 사실혼도 사실관계에 따라 전혼인 법률혼이 사실상 이혼상태에 놓여있는 특별한 사정이 있다면 중혼적 사실혼도 보호할 수 있다. 그러나 판례에서 언급하고 있는 사실상 이혼은 민법 규정에 정한 바 없고, 이제까지 사실상 이혼의 성립요건 및 그 효과에 대하여 논의하고 있는 문헌도 많지 않다. 사실상 이혼을 인정하려면 어떠한 요건을 갖추어야 하고, 어떠한 법률효과가 발생 또는 제한이 되는지에 대하여 연구할 필요성이 있다. 사실상 이혼의 성립에서 살펴보아야 할 것은 부부가 장기간의 별거로 인하여 혼인관계의 실질이 존재하지 않는 경우에 이혼의사의 합치가 있었는가에 따라 다르게 볼 것인가이다. 이와 관련하여 우리 민법상 법률 혼주의와 유책주의 이혼법을 근거로 검토해 보았다.
The purpose of the present study is to investigate the effect of time remaining to use mileage in the notification message from retailers on consumer responses. A total of 577 consumers participated in experiments involving different notification messages of the time remaining to use mileage. Results showed: 1) a significant difference in mileage benefit perception, positive emotion, negative emotion, attitude toward retailers, and repurchase intention according to the remaining time to use mileages, 2) benefit perception positively affected positive emotion and negatively affected negative emotion; positive emotion positively affected and negative emotion negatively affected attitude toward retailers; and attitude positively affected repurchase intention on retailers, and 3) the remaining time to use mileages moderates the relationship between attitude and repurchase intention. Findings highlighted the importance of timing of the message to notify the consumer as to remaining time to use mileage. In the case of a message indicating long remaining time to use mileage, consumers showed more positive responses toward retailers than did consumers who had a message indicating short remaining time to use mileage. These results can be used as guidelines to select the optimal time to send notification messages of remaining time to use mileage in order to generate positive consumer responses.
한국산개구리(Rana coreana)의 난소주기를 파악하기 위해 암컷 성체를 대상으로 gonadosomatic index(GSI)와 난소 내 여포난자의 크기와 난황축적 정도를 기준으로 발달과정을 연중 조사하였다. 난소무게와 GSI는 3월부터 5월까지 가장 낮게 나타났으며 모든 여포난자들은 난황축적 전단계의 상태로 존재하여 난황형성이 중단된 것으로 판단된다. 난소무게와 GSI가 증가하기 시작한 6월의 난소에서는 난황축적 전기단계의 여포난자가 출현하였고 8월에는 난황축적 중기단계와 난황축적 전단계의 여포난자가 존재하여 난소무게와 GSI도 증가하여 나타났다. 이러한 현상은 이 시기에 난황축적 현상이 활발하게 진행되는 것을 의미하며 난소무게와 GSI가 높게 나타난 9월에서 11월까지는 난황축적 중기단계의 여포난자들과 난황축적이 거의 완성된 난황축적 후기단계의 여포난자들이 존재하였다. 동면중인 12월부터 난황축적을 마치고 성장이 완료된 여포난자들이 출현하였으며 2월의 난소에서는 성장이 완료된 여포난자가 전체적으로 존재하여 여포난자의 성장기에는 난소내의 모든 여포난자들이 동시적(synchronized)으로 진행되지 않고 각각의 여포난자에 따라 진행되다가 배란시기에 성장이 완료된 상태를 유지하는 난소주기를 나타내었다.
한국산개구리 (Rana coreana)의 정소주기를 파악하기 위해 수컷 성체를 대상으로 gonadosomatic index (GSI)와 정소 내 생식세포의 변화를 연중 조사하였다. 정소의 세정관내 정자형성은 8월부터 시작되어져 9월의 정소에서 가장 활발하게 진행되었으며 이 시기에 GSI의 값이 가장 컸고 세정관의 단면적도 가장 넓게 나타났다. 2월의 정소에서는 정자배출 후 단계의 세정관들이 출현하였으며 이후 3월~7월까지 일정기간동안 정자형성이 정지되었고 GSI와 세정관의 단면적도 최저치를 나타내었다. 본 결과들로 보아 한국산개구리 수컷의 GSI는 7월에서 8월 사이에 유의하게 변화했고 정자형성과정이 불연속적으로 진행되는 정소주기를 나타내며 번식기는 2월로 확인되었다.
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.
Embryoid bodies (EBs) generated from human embryonic stem cells (hESCs) include spontaneously induced endodermal lineage cells (ELCs). Activin-A plays important roles in the endoderm differentiation of hESCs. Despite studies on the generation of ELCs from hESCs with treatment of Actvin-A, it was unclear for localization and pattern of ELCs by Activin-A during differentiation of hESCs. Accordingly in this study, we knew that Actvin-A increased the cystic EBs formation, including the highly enriched AFP (endoderm lineage specific marker)-expressing cells in the surface of cystic EBs. To induce the EBs formation from undifferentiated hESCs, cells were transferred onto petri-dish and cultured in suspension condition without bFGF removed hESC media (EB media) for 3 days. Next to investigate the effect of Activin-A, EBs were subsequently cultured in EB media supplement with 100 ng/ml Activin-A for 3 days. After 5~7 days of Activin-A treatment, cystic EBs began to appear which increased in numbers reaching ~60% of initially formed EBs over 5 days. Endoderm lineage marker, AFP were highly expressed and specifically localized at the surface region of cystic EBs comparison with normal EBs. We next attached the cystic EBs onto gelatin-coated plates and cultured for 5 days. In the results of real-time PCR and immunocytochemistry analysis, AFP-expressing cells migrated and localized at the outgrowth region of attached cystic EBs. To obtain the AFP-expressing cells of the outgrowth region, we manually isolated by using micro- dissection and cultured them. These cells strongly express AFP over 70% of isolated cells post re-plating. Here, we first showed an expression pattern of specifically localized ELCs by Activin-A during differentiation of hESCs. From this observation, we could highly purified ELCs from undifferentiated hESCs. Taken together, our system will provide a novel and efficient option to generate ELCs from hESCs.
Mesenchymal stem cells (MSCs) have the multipotent capacity and this potential can be applied for obtaining valuable cell types which can use for cell therapy on various regenerative diseases. However, insufficient availability of cellular source is the major problem in cell therapy field using adult stem cell sources. Recently, human embryonic stem cells (hESCs) have been highlighted to overcome a limitation of adult cellular sources because they retain unlimited proliferation capacity and pluripotency. To use of hESCs in cell therapy, above all, animal pathogen free culture system and purification of a specific target cell population to avoid teratoma formation are required. In this study, we describe the differentiation of a mesenchymal stem cell-like cells population from feeder-free cultured hESCs(hESC-MSCs) using direct induction system. hESC-MSCs revealed characteristics similar to MSCs derived from bone marrow, and undifferentiated cell markers were extremely low in hESC-MSCs in RT- PCR, immunostaining and FACS analyses. Thus, this study proffer a basis of effective generation of specialized human mesenchymal stem cell types which can use for further clinical applications, from xenofree cultured hESCs using direct induction system.