• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.95-96
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• 2001

The mutagenic and antimutagenic activities of catechin, hamamelitannin and two proanthocyanidin fractions prepared from the bark of Hamamelis virginia L. - a commonly used medicinal herb - were investigated in a human derived hepatoma (Hep G2) cell line using the single cell gel electrophoresis (SCGE, syn. Comet assay) for the detection of DNA-migration. The cells possess different phase I and phase II enzymes involved in the biotransformation of xenobiotics.(omitted)

• Molecules and Cells
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• 제24권2호
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• pp.268-275
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• 2007

Protein phosphorylation plays a key role in signal transduction in cells. Since phosphoproteins are present in low abundance, enrichment methods are required for their purification and analysis. Chemical derivatization strategies have been devised for enriching phosphoproteins and phosphopeptides. In this report, we employed a strategy that replaces the phosphate moieties on serine and threonine residues with a biotin-containing tag via a series of chemical reactions. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO)-depleted protein extracts prepared from Arabidopsis seedlings were chemically modified for 'biotin-tagging'. The biotinylated (previously phosphorylated) proteins were then selectively isolated by avidin-biotin affinity chromatography, followed by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This led to the identification of 31 protein spots, representing 18 different proteins, which are implicated in a variety of cellular processes. Despite its current technical limitations, with further improvements in tools and techniques this strategy may be developed into a useful approach.

• Restorative Dentistry and Endodontics
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• 제45권2호
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• pp.20.1-20.14
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• 2020

Objectives: To minimize the tooth sensitivity caused by in-office bleaching, many dentists use non-steroidal anti-inflammatory drugs and topical desensitizing gels containing potassium nitrate and sodium fluoride. This study aimed to evaluate the influence of these substances on inflammation and the expression of substance P and calcitonin gene-related peptide in pulp nerve fibers. Materials and Methods: Seventy-two rats were divided into 6 groups as follows: GI, control; GII, only dental bleaching; GIII, only ibuprofen; GIV, ibuprofen administered 30 minutes before and after the bleaching treatment and every 12 hours until the analysis; GV, only topical application of a desensitizing agent; and GVI, topical application of a desensitizing agent before dental bleaching. Placebo gel was applied to the upper left jaw and the bleaching agent was applied to the upper right jaw in all groups. Subsequently, the groups were divided into 3 subgroups based on the time of analysis: 0, 24, and 48 hours after bleaching (n = 8). The rats were euthanized and the maxillae were processed and evaluated by histopathological and immunohistochemical analyses. The data were analyzed using the Kruskal-Wallis test, followed by the Dunn test (p < 0.05). Results: In the bleaching groups, the inflammatory process and expression of neuropeptides decreased over time. The animals in which a desensitizing agent was applied showed better results within 24 hours. Conclusions: The use of a desensitizing agent had positive effects on inflammation and pain-related neuropeptide expression, minimizing the painful effects of dental bleaching treatment.

• 한국양식학회지
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• 제19권1호
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• pp.33-39
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• 2006

Anticoagulant activities of a fermented edible brown alga, Laminaria ochotensis was investigated. L. ochotensis was fermented with 15% sugar (w/v) at $25^{\circ}C$ for 10 weeks. Anticoagulant activity was measured from the supernatant of algal mixture at biweekly intervals up to $10^{th}$ week by activated partial thromboplastin (APTT), prothrombin time (PT) and thrombin time (TT) assay using citrated human plasma. Sample having high APTT activity $(6^{th}\;week)$ was filtered, ethanol precipitated and freeze-dried. The polysaccharide compound having anticoagulant activity was purified by DEAE ion exchange chromatography followed by Sepharose-4B gel filtration chromatography. Anticoagulant activity, polysaccharide concentration, and heparin like activity were determined for the collected fractions by APTT, $phenol-H_2SO_4$, and glycosaminoglycan assay, respectively. The anticoagulant activity assay showed that the activity was increased up to $6^{th}$ week, and decreased thereafter. The concentration of our purified compound was $31.0{\mu}g/ml$ and showed higher APTT activity than commercial heparin. At the same concentration of $31.0{\mu}g/ml$, the heparin showed 186.5 sec activity while our purified compound showed an activity of 386 sec. Single spot on agarose gel electrophoresis showed that the compound was purified and polyacrylamide gel electrophoresis (PAGE) results revealed that the molecular mass of the purified polysaccharide compound was between 60 and 500 kDa. Therapeutic interest of the algal polysaccharide as an anticoagulant has recently been in highlighted. This purified anticoagulant compound from fermented L. ochotensis can be used as a model for anticoagulant agent or could be developed as an anticoagulant agent. This study can be extended to identify the structure and chemical composition of the purified polysaccharide, and to establish a relationship between structure and the function of the identified anticoagulant compounds.

• KSBB Journal
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• 제26권6호
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• pp.517-522
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• 2011

In our previous study, we isolated and characterized a 1-deoxynojirimycin (DNJ)-producing bacterium, Bacillus subtilis MORI, from chungkookjang, a Korean traditional food. B. subtilis MORI was subjected to ${\gamma}$-irradiation and the resulting bacteria were screened for increased DNJ production. A mutant was identified that produced 7.6 times more DNJ and named B. subtilis MORI 3K-85. In this study, the protein profiles of both strains were compared by one-dimensional and two-dimensional gel electrophoresis (1-DE and 2-DE, respectively) under both native and denaturing conditions. The 1-DE native-PAGE and 1-DE SDS-PAGE analyses identified 5 and 7 bands, respectively, that were found at higher concentrations in B. subtilis MORI 3K-85 than in B. subtilis MORI. Similarly, 2-DE analyses identified 20 protein spots which were found at higher concentrations in B. subtilis MORI 3K-85. The peptide mass profiles of these 20 proteins were analyzed by MALDI-TOF and compared with peptide sequences of B. subtilis and B. amyloliquefaciens in the MASCOT database. This screening suggested that three dehydrogenases, an aldolase, a synthetase, an isomerase, a reductase, and a peroxidase are elevated in B. subtilis MORI 3K-85. Based on this data, one or more of the elevated 8 enzymes might be related to the DNJ biosynthetic pathway.

• BMB Reports
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• 제38권3호
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• pp.307-313
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• 2005

Thiobacillus ferrooxidans is one of the most important bacterium used in bioleaching, and can utilize $Fe^{2+}$ or sulphide as energy source. Growth curves for Thiobacillus ferrooxidans have been tested, which show lag, logarithmic, stationary and aging phases as seen in other bacteria. The logarithmic phases were from 10 to 32 hours for Thiobacillus ferrooxidans cultivated with $Fe^{2+}$ and from 4 to 12 days for Thiobacillus ferrooxidans cultivated with elemental sulphur. Differences of protein patterns of Thiobacillus ferrooxidans growing on elemental sulphur and $Fe^{2+}$ separately were investigated after cultivation at $30^{\circ}C$ by the analysis of two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ ionization (MALDI)-Mass spectrometry and ESI-MS/MS. From the 7 identified protein spots, 11 spots were found more abundant when growing on elemental sulphur. By contrast 6 protein spots were found decreased at elemental cultivation condition. Among the proteins identified, cytochrome C have been previously identified as necessary elements of electron-transfering pathway for Thiobacillus ferrooxidans to oxidize $Fe^{2+}$; ATP synthase alpha chain and beta are expressed increased when Thiobacillus ferrooxidans cultivated with $Fe^{2+}$ as energy source. ATP synthase Beta chain is the catalytic subunit, and ATP synthase alpha chain is a regulatory subunit. The function of ATPase produces ATP from ADP in the presence of a proton gradient across the membrane.

• Korean Journal of Acupuncture
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• 제29권2호
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• pp.260-270
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• 2012

Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

• 한국산학기술학회논문지
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• 제9권3호
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• pp.800-806
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• 2008

자연발효된 청국장으로부터 혈전용해활성을 가지는 세균 Bacillus licheniformis HK-12를 농화분리하여, 배양기간 동안 생산된 혈전용해활성을 가지는 단백질에 대해 프로테옴 분석을 실시하였다. B. licheniformis HK-12를 액체 영양배지에 접종하여 얻어진 배양상등액을 피브린 평판법(fibrin plate method)을 사용하여 효소활성을 측정하였다. 그 결과, HK-12의 혈전용해 활성은 대조구인 plasmin보다 약 2.3배정도 높은 활성을 나타내었다. 효소는 배양상등액을 ammonium sulfate 침전, DEAE-cellulose chromatography, Sephadex chromatography 등을 수행하여 분리정제하였으며, 정제된 혈전용해효소의 분자량은 SDS-PAGE를 통해 약 23 kDa로 측정되었다. 배양시간에 따른 HK-6의 세포외 단백질의 변화를 2-D PAGE 분석을 통하여 분석하였다. 그 결과 36시간배양 후에 가장 현저하게 유도된 spot #1을 분리하였으며, MALDI-TOF MS를 이용하여 단백질 동정을 실시한 결과, 유도된 단백질의 아미노산 서열은 $^1EKKIEKYREEEORLK^{15}$으로서, serine protein kinase (PrkA) (AAU22526)로 확인되었다.

• 한국축산식품학회지
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• 제26권2호
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• pp.263-268
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• 2006

최근 들어 병원성 미생물의 저감화를 위하여 기존의 항생제 계열의 항균물질이 아닌 새로운 개념의 신소재 개발이 활발하게 진행 중에 있다. 특히, 이러한 신개념의 병원성 저감화 소재 중 인간의 장내에 존해하는 probiotics 균주의 특성을 이용하여 병원성 미생물을 예방하는 것은 보다 효과적인 방법 중의 하나가 될 수 있을 것으로 판단된다. 본 실험에서는 HT-29 cell을 대상으로 L. acidophilus 균체와 세포 파쇄물을 대상으로 STEC ATCC 43894의 장 상피세포 부착 억제능력을 측정하였다. 10 mg/mL의 세포 파쇄물이 존재하였을 때 $10^9cfu/mL$의 균체가 존재했을 때와 유사한 수준으로 STEC ATCC 43894의 부착 저해 효과가 관찰되었다. 하지만, L. acidophilus A4의 상등액에서는 그 저해 효과가 세포 파쇄물의 $5{\sim}10%$ 정도 수준으로 관찰되어 그 효과는 매우 적은 것으로 판단되었다. 또한, L. acidophilus A4의 세포 파쇄물이 STEC의 부착에 미치는 영향을 관찰하기 위하여 10mg/mL의 세포 파쇄물이 첨가된 배지에서 STEC의 단백질발현 양상을 확인하였다. 각 gel의 image에서 평균적으로 800개의 spot을 관찰할 수 있었으며 이중 2배 이상의 발현차이를 보이는 13개의 spot을 선발하였다. 7개의 spot은 세포파쇄물이 첨가되었을 때 발현이 증가하였으며 3개의 spot은 발현이 감소하였다. 흥미롭게도 3개의 단백질 spot은 세포파쇄물이 존재할 때만 발현되는 것을 확인하였다. 명확하지는 않지만 이러한 L. acidophilus A4의 세포 파쇄물에 존재하는 물질은 (1)STEC의 부착과 관련된 특정 단백질의 발현을 저해하거나 (2)STEC과 장상 피세포에서의 수용체 경합을 통해 부착을 억제하는 것으로 생각된다. 앞으로 이와 관련된 보다 세부적인 작용 메카니즘 연구 및 생화학적연구가 필요할 것으로 판단된다.

• 한국균학회지
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• 제6권2호
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• pp.1-10
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• 1978

검정곰팡이(Aspergillus niger)의 액침배양과 액체표면배양을 통한 동조적 형태분화에 있어서 체외효소인 단백질분해효소, 알파 및 굴루크아밀라제의 신생적 생합성 상황을 연구하였다. 굴루크아미라제는 경자(phialide)가 성숙하는 단계 즉 포자 형성의 전단계에서만 그 활성이 왕성하였다. 단백질분해효소(산성, 중성 및 알칼리성)들은 분생자병의 성장단계에서 활성이 약간 증가하였으나, 경자의 성숙단계에서는 활성이 극히 활발하였다. 알파아밀라제는 경자의 성숙시기와 포자형성기에서 활성이 활발하였으며 그 활성은 장기간 지속 되었다. 알파아밀라제의 활성은 포자형성 기간중 계속 증가하였으므로 신생적으로 생합성된다고 할 수 있으며, 포도당배지에서 많은 량이 생합성되었고, 또 포도당량의 고갈에 즈음하여 그 생합성이 개시되었으므로 이 효소는 구성적 효소이며 이화 대사물의 억제작용(catabolite repression)을 받는 효소라고 할 수 있다. 포리아크릴아미드 젤(polyacrylamide gel)을 이용한 전기영동으로서 포자형성기와 그 전단계의 균체로부터 다양하고 선명한 세포외성 단백질을 분리할 수 있었다. 균사형성기나 포자발아기의 균체로부터는 극소수의 선명치 못한 분리상을 얻었다. C-14 우라실이 균체의 RNA핵산으로 섭취되어 들어가는 비율과 C-14 굴루탐산이 균체 단백질으로 섭취되어 들어가는 비율은 포자형성전기에서 왕성하였으며 포자형성 기간중에는 극히 저조하였다. 알파아밀라제의 신생적 생합성과 포자형성이 일치하는 현상은 유전인자의 표현과정이 내포되는 분화(포자형성)라는 점에서 볼 때 의의와 인과관계가 있을 것으로 사료된다.(1.9 eV)와 $e_g$ $(2.8{\sim}3.0\;eV)$로의 전이 즉, $O^{2-}(2p){\rightarrow}Mn^{4+}(3d)$ 및 $O^{2-}$에서 $Mn^{3+}$ 이온의 $t_{2g}$ (2.3 eV)와 $e_g$ ($3.4{\sim}3.6$ eV)로의 전이 즉, $O^{2-}(2p){\rightarrow}Mn^{3+}(3d)$ 등에 의한 것으로 해석된다. 또한, 1.6, 1.8, 1.9 eV 부근에서 관측된 좁은 에너지 영역의 흡수구조 들은 팔면체 $Mn^{3+}$ 이온 내에서의 d-d 결정장(crystal-field) 전이에 의한 것으로 해석된다. 이러한 흡수구조는 Ni 치환량이 증가함에 따라 그 강도가 감소한다. x = 0.6의 경우 $e_g$ 상태와 관련된 CT 전이구조 들이 $t_{2g}$ 상태와 관련된 전이구조 들에 비하여 큰 폭으로 감소하는데 이것은 Jahn-Teller 효과에 의해서 격자상수가 tetragonal 구조로 확장됨에 따라 $e_g$ 상태와 $O^{2-}(2p)$ 상태 간의 파동함수 중첩이 감소한 것에 기인하는 것으로 해석된다.)의 영향(影響)을 더 많이 받고 있었다. 마. total ginsenosides의 분해반응시(分解反應時)의 활성화(活性化)에너지($E_a$)는 17.7kcal/mole이었고