• 한국약용작물학회지
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• 제14권1호
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• pp.36-40
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• 2006

증숙단계별 고려홍삼의 조직학적 특성을 조사하기 위하여 본 연구를 수행하였다. 시료는 6년근 고려인삼을 60분, 120분동안 증숙한 후 수분함량을 $13{\pm}0.5%$ 까지 건조 후, 수직 수평으로 절단하였다. 조직은 표피, 피층, 목질부로 구분하여 주사전자현미경으로 비교 관찰하였다. 증숙 없이 건조된 시료는 세포막이 유지되고 그 안에 전분입자의 결정성이 유지되어 있었다. 반면에 증숙시간이 연장될수록 호화 후 건조에 따른 수축으로 인하여 조직이 치밀하여지고, 세포조직 사이의 도관, 수지관 등의 동공 및 세포막의 구분이 없어진 것이 특징이었다. 또한 수산화칼슘 족정은 증숙시간 60분 이상에서는 발견되지 않았다. 이러한 원인은 내용성분이 호화 된 후 건조로 인해 조직이 수축되기 때문으로 생각된다. 이로서 증숙단계에 따른 홍삼 조직의 분명한 특성을 비교 관찰할 수 있었다.

• Investigative Magnetic Resonance Imaging
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• 제2권1호
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• pp.73-82
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• 1998

목적 : 간특정 MR 조영제를 이요하여 간세포의 세포막을 통한 물분자의 교환 및 세포막 투과율을 정확히 측정 할 수 있는 MR 기법을 개발하고자 하였다. 대상 및 방법 : 쥐의 간세포를 분리하여 낸 후 NMR 측정을 시도하엿다. 모든 실험은 0.02MHz부터 60 MHz까지 양성자의 Larmor 주파수를 변화시킬 수 있는 IBM형 field cycling relaxometer를 사용하여 시행하였으며 spin-echo 펄스열을 사용하여 T1 자기이완시간을 측정하였다. 전오도의 간특정 조영제인 Gd-EOB-DTPA를 함유하고 있는 간세포 샘플로부터 획득한 T1 데이터를 연속분포 분석법을 사용하여 분석하였으며 이때 이론적 모델로는 Two compartmental exchange 모델을 이용하였다. 결과 : 간세포내의 물분자의 평균 거주시간은 약 250 msec이며 간세포막의 투과율에 대한 최저치는 $(1.3{\pm}0.1){\;}{\times}{\;}10^{-3}cm/sec$ 이었다. 자기이완시간의 연속적인 분포도를 구할 수 있는 CONTIN 분석기법을 적용한 결과 확산적 물분자 교환이 일어남을 밝혔고 이러한 확산적 교환의 정도가 간세포의 경우 세포내 공간에서는 작지 않다는 사실을 규명 할 수 있었다. 결론 : 연속분포 분석기법을 적용하는 경우 Gd-EOB-DTPA는 간세포에서의 물분자의 교환정도 및 세포막의 물분자에 대한 투과율을 측정하는데 매우 유용한 방법임을 확인하였고 간세포에서의 물분자의 교환속도는 적혈구에서의 물분자 교환 속도에 비해 매우 느리다는 사실을 확인하였다. 따라서 조직 특정 조영제는 해당 조직 혹은 세포의 세포막 투과율과 같은 생리학적 정보를 알아낼 수 있는 기능적 조영제로서의 유용성을 입증할 수 있었다.

• Applied Microscopy
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• 제17권1호
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• pp.98-114
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• 1987

Degranulation of the rat peritoneal mast cell induced by intraperitoneal injection of horseradish peroxidase(HRP) was studied using light and electron microscopes. 1. Rat peritoneal mast cells in the Tyrode's buffered salt solution injected control group did not show any particular morphological changes following the specified time course. 2. Under the light microscope, the majority of mast cells observed 10 minutes after HRP injection were nearly the same as those of the control group. However, after 30 minutes, granule densities or staining properties of certain cells began to decrease and these appearances increased gradually until 12 hours after injection, at which time small groups of granules being stained pale-red or pink with toluidine blue were easily identified in the cytoplasm of many cells, and numerous extruded granuleg were scattered around these cells. 3. In the mast cells representing the early stage of degranulation induced by HRP, the electron densities of certain granules decreased as the size enlarged, and perigranular cavities were formed by perigranular membrane expansion. As a result, a thin cytoplasmic septum was formed between the expanded perigranular membrane and the cytoplasmic membrane in the cell periphery, and fusion of the adjacent perigranular membranes was observed in the inner side of the cell. 4. In some mast cells, one or two changes in the peripheral cytoplasmic septum could be seen. One was a focal rupture of the peripheral septum and the other was the formation of a saccule containing one or more vesicles. This saccule was thought to be used for granule-extrusion site and/or material absorptive apparatus judging from the morphological characteristics. 5. As the degranulation proceeded, the granule was extruded from the cell after partial rupture of the peripheral cytoplasmic septum. This phenomenon proceeded to-ward the inner side of the cell through the fused perigranular cavities, and consequently several distinct cavities containing a few unextruded membrane-free granules were formed throughout the cytoplasm after 12 hours. As a rule, the granule-extrusion sites were relatively fewer while the cytoplasmic cavities resulting from degranulation were more numerously observed. Thus, it was thought that the granule-extrusion sites tended to be restricted in the HRP-induced degranulation.

• Journal of Ginseng Research
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• 제16권2호
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• pp.129-137
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• 1992

This study was carried out to investigate the localization of lipids and lipase activity with lipid staining and cytochemical technique in endosperm cells of Panax ginseng C.A. Meyer seed. In endosperm cells of indehiscent seed, protein bodies facing the umbiliform layer are different in electron density during the various degraded processes. Gradually, protein matrix near the cell wall was lysed and electron lucent inclusions appeared on umbiliform layer. The protein body with high electron density and the spherosome with low electron density were observed in endosperm cells. As a result of lipid staining, electron density of spherosome is more intense than those of the protein matrix within the protein body in endosperm cells of indehiscent seed. Free spherical spherosomes within the umbiliform layer have a high electron density. The spherical spherosomes were more electron densed and were uniform in comparison with the cytoplasmic proteinaceous granules in endosperm cells of seed with red seed coat. The major component of spherosome was determined to be lipid. Lipase activity occurs in the spherosome and near the endosperm cell wall facing the umbiliform layer. Cytochemical reaction products of lipase were observed in the spherosome membrane and in the inner regions of spherosome. After protein bodies were digested, lipase activities were observed in free spherosomes and near the cell wall of endosperm cells. Umbiliform layer composing of fibrillized wall and digested materials of the endosperm cell showed a little lipase reaction products.

• Biomolecules & Therapeutics
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• 제27권1호
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• pp.78-84
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• 2019

Cell therapeutic agents for treating degenerative brain diseases using neural stem cells are actively being developed. However, few systems have been developed to monitor in real time whether the transplanted neural stem cells are actually differentiated into neurons. Therefore, it is necessary to develop a technology capable of specifically monitoring neuronal differentiation in vivo. In this study, we established a system that expresses cell membrane-targeting red fluorescent protein under control of the Synapsin promoter in order to specifically monitor differentiation from neural stem cells into neurons. In order to overcome the weak expression level of the tissue-specific promoter system, the partial 5' UTR sequence of Creb was added for efficient expression of the cell surface-specific antigen. This system was able to track functional neuronal differentiation of neural stem cells transplanted in vivo, which will help improve stem cell therapies.

• 농업생명과학연구
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• 제44권2호
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• pp.57-66
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• 2010

자색고구마 추출물의 항산화 효과와 산화적 스트레스로 유도된 PC12 신경세포에 대한 보호효과에 대하여 연구하였다. 자색고구마 추출물의 총 페놀함량은 44.25 mg/g, monomeric anthocyanin 함량은 2,394 mg/L로 나타났다. 자색고구마 추출물의 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis-(3-ethylben-zthiazoline-6-sulfonic acid) (ABTS) radical 소거활성, ferric reducing/antioxidant power (FRAP) 및 환원력은 농도 의존적으로 항산화 활성이 증가하였다. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazoliumbromide} reduction assay를 이용하여 자색고구마 추출물의 신경세포 보호효과를 측정한 결과, 세포 생존율이 두드러지게 증가하는 것으로 나타났다. 산화적 스트레스는 신경세포막 손상 정도를 증가시키기 때문에 lactate dehydrogenase (LDH) release assay와 neutral red uptake assay를 이용하여 세포막 손상 보호효과를 조사한 결과 자색고구마 추출물 처리구는 대조구에 비하여 산화적 스트레스로 유도된 세포막 손상 보호효과가 농도 의존적으로 나타났다. 따라서 자색고구마 추출물은 천연 항산화 소재 및 알츠하이머성 치매와 같은 신경퇴행성 질환의 예방 소재로서의 활용 가능성이 기대된다.

• The Korean Journal of Physiology and Pharmacology
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• 제13권1호
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• pp.39-47
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• 2009

Gaegurin 4(GGN 4), an antimicrobial peptide isolated from a Korean frog, is five times more potent against Gram-positive than Gram-negative bacteria, but has little hemolytic activity. To understand the mechanism of such cell selectivity, we examined GGN4-induced $K^+$ efflux from target cells, and membrane conductances in planar lipid bilayers. The $K^+$ efflux from Gram-positive M. luteus(2.5 ${\mu}g/ml$) was faster and larger than that from Gram-negative E. coli(75 ${\mu}g/ml$), while that from RBC was negligible even at higher concentration(100 ${\mu}g/ml$). GGN4 induced larger conductances in the planar bilayers which were formed with lipids extracted from Gram-positive B. subtilis than in those from E. coli(p<0.01), however, the effects of GGN4 were not selective in the bilayers formed with lipids from E. coli and red blood cells. Addition of an acidic phospholipid, phosphatidylserine to planar bilayers increased the GGN4-induced membrane conductance(p<0.05), but addition of phosphatidylcholine or cholesterol reduced it(p<0.05). Transmission electron microscopy revealed that GGN4 induced pore-like damages in M. luteus and dis-layering damages on the outer wall of E. coli. Taken together, the present results indicate that the selectivity of GGN4 toward Gram-positive over Gram-negative bacteria is due to negative surface charges, and interaction of GGN4 with outer walls. The selectivity toward bacteria over RBC is due to the presence of phosphatidylcholine and cholesterol, and the trans-bilayer lipid asymmetry in RBC. The results suggest that design of selective antimicrobial peptides should be based on the composition and topology of membrane lipids in the target cells.

• 대한임상검사과학회지
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• 제39권2호
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• pp.122-127
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• 2007

This study is a report about a specific patient whose primary stomach adenocarcinoma metastasized to uterine cervix adenocarcinoma. A thirty-nine year old female patient was initially diagnosed as having metastatic adenocarcinoma in the supraclavicular lymph node. Upon further examination, she was diagnosed with stomach adenocarcinoma. 8 months later, a cervix punch biopsy was performed. The stains used for examination were H&E stain, PAS stain, Alcian blue stain, Mucicarmine stain, Papanicolaou's (Pap.) stain, and as immunohistochemical stains, cytokeratin 7 and 20 were done. In the H&E stain, the tumor cells showed prominent and eccentric nuclei, thin nuclear membrane in abundant mucous cytoplasm, and cylinder shape. In the PAS stain, intracytoplasmic mucin vacuoles were stained with pink, and in Alcian blue and Mucicarmine stains, intracytoplasmic mucin vacuoles were stained with blue and red. As in the above results, she was diagnosed with undifferentiated adenocarcinoma. As found on the cytologic smear preparation of the uterine cervix stained by Papanicolaou's stains, the background was relatively clear, the number of malignant cells was relatively low, and large and eccentric nuclei in abundant cytoplasm were observed. Upon observing the tissue preparation of the uterine cervix biopsy by H&E stain, a clear background, large and eccentric nuclei, and a signet ring cell types were observed, and the number of malignant cells were fewer than in the primary uterine cervix adenocarcinoma. The vacuoles in cytoplasm were observed. The nuclear membrane and chromatin were thick and very rough, and upon observation by cytokeratin 7 and 20 of immunohistochemical stain, the tumor cells indicated a positive rate of 70% and 20%, respectively. According to these results, also she was diagnosed with metastasized uterine cervix adenocarcinoma. In summary of the results of pathologic findings on stomach biopsy and cytologic, histopathologic, and immunohistochemical finding on uterine cervix biopsy, the adenocarcinoma of her uterine cervix could assert the adenocarcinoma of signet ring cell type that was metastasized from the primary undifferentiated adenocarcinoma in stomach.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.55-64
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• 2012

Peroxiredoxin II (Prdx II; a typical 2-Cys Prdx) has been originally isolated from erythrocytes, and its structure and peroxidase activity have been adequately studied. Prdx II has been reported to protect a wide range of cellular environments as antioxidant enzyme, and its dysfunctions may be implicated in a variety of disease states associated with oxidative stress, including cancer and aging-associated pathologies. But, the precise mechanism is still obscure in various aspects of aging containing ovarian aging. Identification and relative quantification of the increased proteins affected by Prdx II deficiency may help identify novel signaling mechanisms that are important for oxidative stress-related diseases. To identify the increased proteins in Prdx $II^{-/-}$ mice, we performed RBC comparative proteome analysis in membrane fraction and cytosolic fractions by nano-UPLC-$MS^E$ shotgun proteomics. We found the increased 86 proteins in membrane (32 proteins) and cytosolic (54 proteins) fractions, and analyzed comparative expression pattern in healthy RBCs of Prdx $II^{+/+}$ mice, healthy RBCs of Prdx $II^{-/-}$ mice, and abnormal RBCs of Prdx $II^{-/-}$ mice. These proteins belonged to cellular functions related with RBC lifespan maintain, such as cellular morphology and assembly, cell-cell interaction, metabolism, and stress-induced signaling. Moreover, protein networks among the increased proteins were analyzed to associate with various diseases. Taken together, RBC proteome may provide clues to understand the clue about redox-imbalanced diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제16권1호
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• pp.71-77
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• 2012

Mitochondrial dynamics and distribution is critical for their role in bioenergetics and cell survival. We investigated the consequence of altered fission/fusion on mitochondrial function and motility in INS-1E rat clonal ${\beta}$-cells. Adenoviruses were used to induce doxycycline-dependent expression of wild type (WT-Mfn1) or a dominant negative mitofusin 1 mutant (DN-Mfn1). Mitochondrial morphology and motility were analyzed by monitoring mitochondrially-targeted red fluorescent protein. Adenovirus-driven overexpression of WT-Mfn1 elicited severe aggregation of mitochondria, preventing them from reaching peripheral near plasma membrane areas of the cell. Overexpression of DN-Mfn1 resulted in fragmented mitochondria with widespread cytosolic distribution. WT-Mfn1 overexpression impaired mitochondrial function as glucose- and oligomycin-induced mitochondrial hyperpolarization were markedly reduced. Viability of the INS-1E cells, however, was not affected. Mitochondrial motility was significantly reduced in WT-Mfn1 overexpressing cells. Conversely, fragmented mitochondria in DN-Mfn1 overexpressing cells showed more vigorous movement than mitochondria in control cells. Movement of these mitochondria was also less microtubule-dependent. These results suggest that Mfn1-induced hyperfusion leads to mitochondrial dysfunction and hypomotility, which may explain impaired metabolism-secretion coupling in insulin-releasing cells overexpressing Mfn1.