Epithelial-mesenchymal interaction is well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular micro-environment which provide a epithelial-mesenchymal interaction. The aims of this study were to develop and evaluate the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted human normal oral kertinocyte(NHOK) and immortalized human oral keratinocytes(IHOK) by histological and immunohistochemical analysis. The results were as follows; 1. Best condition of three dimensionally reconstituted IHOK & HaCaT cells are 14 days air-exposure cultivation, 3 days of submerged state, and dermal equivalent consisting type I collagen and IGF cells. 2. In comparison to IHOK, there was better preservation of the overall epidermal strucutures in oral cracioma cells (HN30) & HaCaT cells by organotypic cultures. But orgnaotypic co-culture of the normal keratinocyte showed the thinnest epithelial layer formation. 3. PCNA was detected primarily in the basal layer of normal mucosa and NHOK, whereas was shown throught the epithellium except surface layer of IHOK cells, and it's expression was similar to that of CIS of biopsied patient's tissue 4. Involucrin is expressed in the upper layer of oral mucosa and NHOK raft, but staining for involucrin was induced in the IHOK rafts indicating differentiation is incomplete, and the staining pattern in the IHOK raft was not uniform. 5. Normal oral keratinocyte raft showed weak immunostaining for p53, and p53 expression of IHOK raft increased rahter than in NHOK. In organotypic cultures of normal cells and IHOK, p53 expression was restricted to the proliferative part of epithelium. This is consistent with expression pattern in biopsy specimens of the normal and CIS tissue. 6. In artifically reconstructed NHOK, the pattern of keratin staining showed both similarity and differences from that of intact normal mucosa. An obvious difference was increased expression of CK10 & CK19, and decreased expression of CK6 in a reconstructed NHOK rather than in normal mucosa, and similar expression was in CK4 and CK16. 7. CK19 & CK16 were strongly positive in HPV immortlaized keratinocyte rafts rahter than in NHOK, an indicator of premalignant or malignant changes, while CK10 & CK6 were decreased, and organotypic cultures of IHOK express was similar keratin expression as epithelial dysplasia or CIS tissue. These results suggest that three-dimensional organotypic co-culture of normal oral & immortalized keratinocytes with dermal equivalent consisting type I collagen and fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. Thus this organotypic system can be used for studing mechanism of epitehlial-mesenchymal interaction in particular regulating epidermal diffenentiation and morphogenes

We used three-dimensional Matrigel culture system to examine the morphognesis of normal and malignant salivary glands cell in vitro including acinar cells(AC), myoepithelial cell(MC), salivary gland adenocarcinoma cells(SGT), mucoepidermoid carcinoma cells(MEC), and immortalized human salivary gland cells(HSG). For this purpose, normal and salivary gland tumor cells cultured in 3-D Matrigel, and characterized histologically and immunohistochemically, compared with same cells grown on monolayer culture and patient tissue from biopsy. 1. In three-dimensional Matrigel culture, HSG cells form acinar structure, SGT cells shows duct like structure, and other AC, MC and MEC cells dont' form any structure , and their morphology was different from that of monolayer cells. 2. Matrigel involved cell proliferation at a similar pattern to cells on plastic monolayer cell cultures, and monolayer cell revealed higher cell viability than that of Matrigel cultured cells. 3. All salivary glands cells on Matrigel or monolayer showed strong PCNA expression, and there is no expression difference in these cells. But some cells including myoepithelial cells in normal and salivary gland tumor tissue showing PCNA lavelling, so there is PCNA expression difference among normal and tumor tissue cells. 4. Actin expression was noted in AC cells on Matrigel, were rare expressed in the other cells except in MEC cells, and was present in myoepithelial cell and ductal cells of normal gland tissue. There is actin expression difference between tissue and cultured cells . 5. S-100 immunoreaction was moderateively positive in MC cells of monolayer culture, myoepithelial cells of normal tissue and pleomorphic adenoma, all cancer cells of mucoepidermoid carcinoma tissue, but significantly decreased in all salivary cells on Matrigel. 6. TGase 2 expression was prominent in MC cells of monolayer and Matrigel cultured, in myoepithelial cells of normal gland and pleomorphic adenoma, epidermoid cells of mucoepidermopid carcioma, and strong reaction in MEC and AC cells of monolayer and Matrigel cultured. 7. Expression of CK in monolayer culture showed strong reaction to CK6 in all sailvary gland cells, and mild reaction to CK10 and CK16 for all salivary cells, CK16 and CK19 expression in monolayer culture was similar to that of Matrigel culture. 8. CK6 and CK10 expression was strongest in AC and MC cells on Matrigel, and CK 4 was negative reaction in AC, SGT, MEC cells, strong reaction in MC cells but mild in SGT cells on Matrigel. Expression of CK was rare in HSG cells compared with other salivary gland cells, CK16 was prominent in SGT cells, CK10 and CK16 showed strongest expression in MEC cells of Matrigel. 9. Monolayer culture of HSG cell shwoing strong reaction to CK6, moderate to CK19 and mild to the others CK, but 3D cultured HSG cells reveal mild expression to CK16, and rare to others CK, intercallated duct in normal gland tissue showing strong to CK19, and mild to the others Ck, so there are CK expression difference in tissue, monolayer and 3-D cultured cells. 10. Monolayer culture of MEC cells represent strong reaction to CK6, mild to other CK, 3-D cells showing increased CK expression including CK6, epidermoid cells and intermediate cells in mucoepidermoid carcinoma tissue reveal positive to CK6 and CK16, mucous cell positive to CK10 and CK19, so Matrigel showed similar CK pattern compared to mucoepidermoid carcinoma tissue rather than monolThese data indicate that the interaction of salivary gland cells with basement membrane is an important factor in salivary gland development and cytodifferentiation, so this model system will be useful to study acinar or ductal differentiation in vitro.ayer cultred.

Technology used by human beings has developed drastically over the years. Although people enjoy affluent lives as a result of this development, the depletion of resources has brought about a variety of environmental problems such as emission of fine dust, treatment of waste water, and global warming. Although studies on environmental pollution are being conducted continuously, there are a limited number of studies that analyze research trends from quantitative and qualitative perspectives. In order to examine the current research landscape, we employed Scopus to combine research interest in environmental science with bibliographic analysis. Among 74,089 papers published in 57 journals of environmental science, 3,212 papers were published by Korean authors and citations per publications and Field-Weighted Citation Impact (FWCI) of those papers were 7.3 and 1.0, respectively. By assessing the bibliometric indicators in the field of environmental science, this study provides insight into research trends and related data to aid researchers in developing research strategies.

본 연구는 제주 자생 식물들의 항산화 활성을 측정함으로써 천연항산화 소재로의 활용 가능성이 있는 유망 자원을 발굴하 고자 하였다. 제주도에서 자생하는 식물 11종을 대상으로 총 폴 리페놀 화합물 함량 측정 및 항산화 활성을 측정하였다. 항산화 기능이 좋은 식물 8종을 우선 선별하여 유기용매 분획을 실시하 고 같은 실험을 농도별로 진행하여 가능성이 있는 유망 후보를 선별 하였다. DPPH free radical 소거 활성은 구실잣밤나무의 ethyl acetate 층과 애기달맞이의 buthanol 층에서 각각 IC50값 이 1.6 ㎍/㎖, 2.4 ㎍/㎖로 좋은 활성을 나타내었다. Nitric oxide 생성 저해 활성은 밤나무의 ethyl acetate 층에서 강한 저 해율 보였다. Xanthine oxidase의 억제 효과는 밤나무의 ethyl acetate 층에서 IC50 값이 16 ㎍/㎖로 우수한 활성을 나타내었 다. Superoxide radical 소거 효과는 구실잣밤나무의 ethyl acetate 층에서 IC50 7 ㎍/㎖로 좋은 활성을 나타내었고, hydroxyl radical 소거 활성은 구실잣밤나무의 buthanol 층에서 76%로 우수한 활성을 나타냄을 확인 할 수 있었다. 밤나무를 비롯한 항 산화 효과가 우수한 식물종에 대해 단일 물질에 대한 분리 작업 과 함께 항염증이나 항암, 항노화 등의 실험이 더 깊이 있게 진 행된다면 새로운 천연물 유래 생리 활성 물질로서 학술 및 산업 적 활용이 가능할 것으로 기대된다.

EMS was commonly used to induce mutations for various organisms, causing nucleotides to mispair with their complementary bases. So, chemical mutagenesis has become the best method for inducing mutations in genetic studies. Simple PCR-based detection and high-throughput technologies helped to screen and identify mutations. Degenerate oligonucleotide primed PCR (DOP-PCR) became getting attention for mutation survey because the requirement of sequence information and high cost for designing primers could be diminished. Also, high-throughput sequencing instruments, such as GS-FLX, allowed characterization of nucleic acids and massive mutant analysis. A total of 6,696 aligned pairs for Sinpaldalkong 2 vs. SS2-2 and 6,935 for Sinpaldalkong 2 vs. 25-1-1 were formed for mutation detection. A mutation every 437 bp in SS2-2 and every 402 bp in 25-1-1 was observed. About 2/3 of a total of mutations were single base variation in both comparisons. Mutated and non-mutated fragments from SS2-2 and 25-1-1 were distributed on all LGs. The 25-1-1 had more mutations than SS2-2 compared with their wild type, Sinpaldalkong 2. Local compositional bias was also observed around the mutated G. Our modified DOP-PCR primers were successfully amplified and their amplicons were located on randomly but somewhat targeted regions of soybean genome.