• The Korean Journal of Physiology and Pharmacology
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• 제14권6호
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• pp.385-390
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• 2010

Excessive extracellular matrix (ECM) accumulation is the main feature of chronic renal disease including diabetic nephropathy. Plasminogen activator inhibitor (PAI)-1 is known to play an important role in renal ECM accumulation in part through suppression of plasmin generation and matrix metalloproteinase (MMP) activation. The present study examined the effect of PAI-1 antisense oligodeoxynucleotide (ODN) on fibronectin upregulation and plasmin/MMP suppression in primary mesangial cells cultured under high glucose (HG) or transforming growth factor (TGF)-${\beta}1$, major mediators of diabetic renal ECM accumulation. Growth arrested and synchronized rat primary mesangial cells were transfected with $1\;{\mu}M$ phosphorothioate-modified antisense or control mis-match ODN for 24 hours with cationic liposome and then stimulated with 30 mM D-glucose or 2 ng/ml TGF-${\beta}1$. PAl-1 or fibronectin protein was measured by Western blot analysis. Plasmin activity was determined using a synthetic fluorometric plasmin substrate and MMP-2 activity analyzed using zymography. HG and TGF-${\beta}1$ significantly increased PAI-1 and fibronectin protein expression as well as decreased plasmin and MMP-2 activity. Transient transfection of mesangial cells with PAI-1 antisense ODN, but not mis-match ODN, effectively reversed basal as well as HG- and TGF-${\beta}1$-induced suppression of plasmin and MMP-2 activity. Both basal and upregulated fibronectin secretion were also inhibited by PAI-1 antisense ODN. These data confirm that PAI-1 plays an important role in ECM accumulation in diabetic mesangium through suppression of protease activity and suggest that PAI-1 antisense ODN would be an effective therapeutic strategy for prevention of renal fibrosis including diabetic nephropathy.

• Nutrition Research and Practice
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• 제4권1호
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• pp.23-29
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• 2010

Poly-gamma-glutamic acid ($\gamma$-PGA) is a mucilaginous and biodegradable compound produced by Bacillus subtilis from fermented soybeans, and is found in the traditional Korean soy product, cheongkukjang. This study was carried out to evaluate the effects of $\gamma$-PGA from a food source on the concentration of the neurotransmitter GABA and its metabolic precursor glutamate in diet-induced obese rats. Eight-week old male Sprague-Dawley rats (n=60) were used. The rats were divided into two groups and obesity was induced by providing either a 10% control fat or 45% high fat diet for 5 weeks. The rats were then blocked into 6 groups and supplemented with a 0.1% $\gamma$-PGA diet for 4 weeks. After sacrifice, brain and serum GABA and glutamate concentrations were analyzed by high performance liquid chromatography with fluorometric detection. The rats fed the high fat diet had significantly increased body weights. $\gamma$-PGA supplementation significantly increased serum concentrations of glutamate and GABA in the control fat diet groups while this effect was not found in the high fat groups. In the brain, glutamate concentrations were significantly higher in the $\gamma$-PGA supplemented groups both in rats fed the normal and high fat diets than in the no $\gamma$-PGA controls. GABA concentrations showed the same tendency. The results indicated that $\gamma$-PGA intake increased GABA concentrations in the serum and brain. However, the effects were not shown in obese rats.

• The Korean Journal of Physiology and Pharmacology
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• 제18권6호
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• pp.497-502
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• 2014

Extracellular nicotinamide adenine dinucleotide (NAD) cleaving activity of a particular cell type determines the rate of the degradation of extracellular NAD with formation of metabolites in the vicinity of the plasma membrane, which has important physiological consequences. It is yet to be elucidated whether intact human neutrophils have any extracellular NAD cleaving activity. In this study, with a simple fluorometric assay utilizing $1,N^6$-ethenoadenine dinucleotide (etheno-NAD) as the substrate, we have shown that intact peripheral human neutrophils have scant extracellular etheno-NAD cleaving activity, which is much less than that of mouse bone marrow neutrophils, mouse peripheral neutrophils, human monocytes and lymphocytes. With high performance liquid chromatography (HPLC), we have identified that ADP-ribose (ADPR) is the major extracellular metabolite of NAD degradation by intact human neutrophils. The scant extracellular etheno-NAD cleaving activity is decreased further by N-formyl-methionine-leucine-phenylalanine (fMLP), a chemoattractant for neutrophils. The fMLP-mediated decrease in the extracellular etheno-NAD cleaving activity is reversed by WRW4, a potent FPRL1 antagonist. These findings show that a much less extracellular etheno-NAD cleaving activity of intact human neutrophils compared to other immune cell types is down-regulated by fMLP via a low affinity fMLP receptor FPRL1.

• 한국수산과학회지
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• 제29권2호
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• pp.165-173
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• 1996

무각류의 와편모조인 C. polykrikoides에 의한 적조는 맹독성으로 알려져 있으며, 이로 인한 어패류의 폐사로 매년 막대한 손실을 초래하고 있어, 어패류 폐사의 원인 독성분을 규명하고, 유용 조류로서 활용하기 위한 기초 자료를 얻을 목적으로 93년 10월 경남 통영군 욕지도 인근에서 발생한 적조시 채취한 천연 조체를 시료로 수종의 생리활성을 검색하였다. MeOH 추출물의 마우스 치사 독성은 없었으며 (지용성 획분 : 50mg, 수용성 획분 : 10mg), 미량 형광 HPLC 분석에서도 마비성이나 설사성 패류독은 검출되지 않았다. 또한, 5mg/ml의 농도에서는 어독성을 나타나지 않았으며, 항균성도 없었다 (10mg/disc). P-388 종양 세포에 대하여는 지용성 획분이 $180{\mu}g/m1$의 농도에서 $20\%$의 생육 저지활성을 나타내었고, 아미노산 분석에서 2개의 미확인 아미노산이 검출되었다. 용혈활성을 나타내는 성분은 지용성, 수용성 양 획분에 모두 존재하였으며, 지용성 획분중의 성분으로서는 EPA와 DHA가 동정되었고, 이들이 총지방산의 $40.6\%$를 차지하여 고도 불포화 지방산 자원으로서 이용 가능성을 시사하였다.

• KSBB Journal
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• 제20권4호
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• pp.291-298
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• 2005

본 연구는 재조합 대장균과 S.cerevisiae의 발효공정에서 형광스펙트럼 데이터를 수집하였으며, SOM을 이용하여 형광스펙트럼 데이터를 특정 그룹으로 분류하고 발효공정을 분석하고자 하였다. 배출가스 내 이산화탄소농도와 세포농도 같은 공정변수들은 SOM 알고리즘으로부터 얻은 분산 및 정규화된 가중치들과 좋은 연관성을 나타내었다. 전체 스펙트럼 데이터의 분류는 생물공정 모델링을 위한 매우 중요한 단계인데 그 이유는 몇몇 여기파장과 방출파장의 유의한 조합들이 전체영역의 스펙트럼 데이터로부터 추출되기 때문이다. 예를 들면, 본 연구에서 SOM을 이용하여 추출한 98개의 스펙트럼 데이터의 예제들은 부분최소자승법이나 감독신경망 (supervised neural network)을 이용한 공정의 모델링에 사용될 수 있다.

• 대한의생명과학회지
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• 제21권1호
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• pp.9-14
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• 2015

Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

• 한국환경과학회지
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• 제15권8호
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• pp.727-735
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• 2006

In this study, the PHYTO-PAM-fluorometric method was used to evaluate the ETR$_{max}$ in terms of sensitivity to DIN/DIP against 14 microalgae: Prorocentrum micans, Heterocapsa triquetra, Gymnodinium impudicum, Cymnodinium catenatum, Amphidinium caterae, Chlorella vulgaris, Chroococcus minutus, Microcystis aeruginosa, Chlorella ellipsoidea, Nannochloris oculata, Oocystis lacustris, Chroomonas salina, Gloeocystis gigas, and Prymnessium parvum. We found that P. micans, H. triquetra, and A. caterae exposed to the maximum level of DIN/DIP were significantly smaller in the ETR$_{max}$ than that of the minimum and moderate mixture. Unlikely the ETR$_{max}$, the initial slope alpha was not significantly different at the level of 60 DIN/DIP. In G. catenatum, the moderate levels of 15 and 20 in DIN/DIP were found to be significantly different from the ETR$_{max}$ at Chl-Ch4. Gymnodinium impudicum had a higher value than that of the ETR$_{max}$ than that of dinoflagellates used in this study, ranging from 306.1 (Ch4, DIN/DIP: 10) to 520.1 (Ch4, DIN/DIP: 30). The ETR$_{max}$ value obtained from other microalgae was similar to C. impudicum at any of the ratios of DIN/DIP and channels. Consequently, the influence of offshore water current assures us of the suppression of photosynthesis and electron transport rate in dinoflagellates. Gymnodinium impudicum has not been researched in the area of red tides in Korea, but it will be enough to creat the massive algal blooms in the future because of higher potential photochemical availability.

• 한국수산과학회지
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• 제24권3호
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• pp.193-202
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• 1991

4종류의 남해안 해면 Galichondria sp., H. okadai, H. japonica 및 Haliclona permollis 중의 항균, 항곰팡이 물질을 paper disk plate법으로 검색하였다. 항균물질은 4종류의 해면에 모두 존재하였으나 항곰팡이 물질은 Halichondria sp.와 H. okadai의 $CH_2Cl_2$와 BuOH구에만 나타났다 이들 두 종류의 해면(184kg)을 아세톤으로 추출하여 각종 column을 이용하여 정제 후 핵자기공명 및 질량분석 등 각종 기기분석을 통하여 benzoic acid, okadaic acid(OA), dinophysistoxin-1(DTX1)의 3성분을 항균, 항곰광이 물질로 동정하였다. 한천 배지상에서의 OA및 DTX1의 미생물 생육저지능력은 비슷하였으며 특히 곰팡이에 대하여 저지효과가 크게 나타났다. 9-Anthryldiazomethane(ADAM)을 이용한 형광 HPLC법을 개발하여 해면중의 OA 및 DTX1을 분석한 결과 이들 성분은 H. okadai 및 Halichondria sp. 에만 존재하였다. 또한, 이들 두 종류의 해면중의 함량은 습중양 1kg당 OA가 $550{\~}600{\mu}g,\;DTX1$이 $400{\~}490{\mu}g$이었다.

• 한국식품영양과학회지
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• 제7권2호
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• pp.35-42
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• 1978

General components of comfrey collected during the period of July-September 1978 in Pusan area were investigated with a view to determine its possible value as a foodstuff by means of titrimetry, chromometric analysis and fluorometric analysis. As a result, the following data were obtained. 1) The contents of moisture, crude lipid, crude protein and carbohydrates of the comfrey were similar to those of other green vegetables such as spinach, crowndaisy, lettuce, leek and mallow. 2) The inorganic Ca, P and Fe of the comfrey were fairly greater in contents than those of other green vegetables. Compared with spinach, for instance, there were 4 times as much Ca and about 2 times as much Fe in it, respectively. 3) The contents of vitamins of the comfrey were also found in an amount much greater than those of other green vegetables. V-A of the comfrey, for instance, were 6 times greater in content than that of lettuce, and about 2 times greater than that of crowndaisy, leek and mallow, respectively. It also showed a greater content of V-A than even that of spinach, known as a vegetable most rich in V-A. V-B of the comfrey showed a little lower level of content than that of spinach but showed about the same level of content as that of leek and lettuce. V-B of the comfrey was 6 to 8 times greater in content than that of lettuce and leek. The content of niacin of the comfrey was also found much greater than those of other foodttuffs. V-C of the comfrey compared 3.5 times greater with that of lettuce, 2 times greater with that of mallow, and generally a higher level with those of other green vegetables. The above findings seem to firmly establish the food scientific value of the comfrey. Furthermore, the vegetable is very prolific and grows easily. If the general public is made aware of these facts, the vegetable is thought to contribute not a little to their dietary life as a nutritious foodstuff.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.12-25
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• 2003

L-camitine ($\beta$ -hydroxy-${\gamma}$ -trimethyl-ammoniumbutyric acid) is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-camitine, we investigated the effects of in vitro MMP inhibition and activity and expression of UVA-induced MMP 1 in human skin fibroblasts. Fluorometric assays of the proteolytic activities of MMP-l were performed using fluorescent collagen substrates. ELISA (enzyme linked immuno sorbent assay), gelatin-substrate zymography, and RT-PCR ELISA techniques were used for the effects of L-camitine on MMP expression and activity, MMP mRNA expression in UVA irradiated fibroblast. L-camitine inhibited the activities of MMP-l in a dose-dependent manner and the $IC_{50}$/ values calculated from semi-log plots were 2.45mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP expression was reduced 40% by treated with L-carnitine, and MMP-l mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibition the MMP activity, regulation of MMP expression in protein and mRNA level. All these results suggest that L-carnitine may be useful as new anti-aging cofactor for protection against UVA induced MMP expression and activity.