• BMB Reports
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• 제36권3호
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• pp.258-264
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• 2003

This study examined the effect of dietary polyunsaturated fatty acids (PUFA) that were supplemented with vitamin E on lipid peroxidation, glutathione-dependent detoxifying enzyme system activity, and lipogenic fatty acid synthase (FAS) expression in rat liver. Male Sprague-Dawley rats were fed semipurified diets containing either 1% (w/w) corn oil or 10% each of beef tallow, corn oil, perilla oil, and fish oil for 4 wk. Alpha-tocopherol was supplemented in perilla oil (0.015%) and fish oil (0.019%). Hepatic thiobarbituric acid reactive substances, an estimate of lipid peroxidation, were not significantly different among the dietary groups. The glutathione peroxidase, glutathione reductase, and glutathione S-transferase activities were all elevated by the polyunsaturated fats, especially fish oil. The activity of FAS was reduced in the polyunsaturated fat-fed groups in the order of fish oil, perilla oil, and corn oil. The mRNA contents decreased in rats that were fed the 10% fat diets, particularly polyunsaturated fats, compared with the rats that were fed the 1% corn oil diet. Similarly, the inhibitory effect was the greatest in fish oil. These results suggest that lipid peroxidation can be minimized by vitamin E; PUFA in itself has a suppressive effect on lipogenic enzyme.

• Biomolecules & Therapeutics
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• 제26권4호
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• pp.409-416
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• 2018

Vasicinone, a quinazoline alkaloid from Adhatoda vasica Nees. is well known for its bronchodilator activity. However its anti-proliferative activities is yet to be elucidated. Here-in we investigated the anti-proliferative effect of vasicinone and its underlying mechanism against A549 lung carcinoma cells. The A549 cells upon treatment with various doses of vasicinone (10, 30, 50, $70{\mu}M$) for 72 h showed significant decrease in cell viability. Vasicinone treatment also showed DNA fragmentation, LDH leakage, and disruption of mitochondrial potential, and lower wound healing ability in A549 cells. The Annexin V/PI staining showed disrupted plasma membrane integrity and permeability of PI in treated cells. Moreover vasicinone treatment also lead to down regulation of Bcl-2, Fas death receptor and up regulation of PARP, BAD and cytochrome c, suggesting the anti-proliferative nature of vasicinone which mediated apoptosis through both Fas death receptors as well as Bcl-2 regulated signaling. Furthermore, our preliminary studies with vasicinone treatment also showed to lower the ROS levels in A549 cells and have potential free radical scavenging (DPPH, Hydroxyl) activity and ferric reducing power in cell free systems. Thus combining all, vasicinone may be used to develop a new therapeutic agent against oxidative stress induced lung cancer.

• 생명과학회지
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• 제19권11호
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• pp.1529-1537
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• 2009

L-arginine 구조유사체인 L-canavanine의 인체 급성백혈병 Jurkat T 세포에 대한 apoptosis 유도활성이 단백질 티로신키나아제 $p56^{lck}$에 어떻게 조절되는지를 규명하기 위해 $p56^{lck}$를 발현하는 Jurkat T 세포주 E6.1과 $p56^{lck}$-결손 Jurkat T 세포주 JCaM1.6에 있어서 L-canavanine의 세포독성, L-canavanine에 의해 유도되는 apoptotic DNA fragmentation 및 apoptotic sub-$G_1$ peak를 비교하여 본 바, $p56^{lck}$-negative JCaM1.6 세포가 $p56^{lck}$-positive E6.1 세포에 비해 L-canavanine의 apoptotis 유도활성에 훨씬 더 민감한 것으로 나타났다. 이러한 $p56^{lck}$-negative JCaM1.6 세포의 민감성은 JCaM1.6 세포에 $p56^{lck}$ 유전자를 transfection시켜 발현시키면 현저히 감소되었다. L-Canavanine에 의해 유도되는 apoptosis관련 현상들을 $p56^{lck}$-stable transfectant인 JCaM1.6/lck 세포와 empty vector-transfectant 인 $p56^{lck}$-negaive JCaM1.6/vector 세포에서 Western blot analysis로 비교한 결과, L-canavanine에 의해 유도되는 mitochondrial membrane potential (${\Delta\Psi}m$)의 감소, caspase-9, -8, -7 및 -3의 활성화, 그리고 PARP 및 $PLC{\gamma}$-1의 분해가 JCaM1.6/vector 세포에 비해 JCaM1.6/lck 세포에서 더 약하게 나타났다. JCaM1.6/lck 세포를 2.5 mM L-canavanine으로 처리한 다음 세포 내 $p56^{lck}$ kinase 활성의 변화를 $\alpha$-casein을 기질로 하여 시간 별로 측정한 결과, L-canavanine의 처리 후 15분만에 $p56^{lck}$ kinase의 활성이 약 1.6배 증가되었으며 이후 6시간 동안은 약 1.3~1.4 배정도 증가된 수준으로 kinase 활성이 유지되는 것으로 확인되었다. L-Canavanine에 의한 apoptosis의 개시에 Fas/FasL 상호작용이 관련되는지를 규명하기 위해 FADD-negative Jurkat T 세포주 I2.1, caspase-8-negative Jurkat T 세포주 I9.2 및 wild-type Jurkat T 세포주 A3에 대한 L-canavanine의 세포독성을 비교한 결과, A3와 I2.1 세포의 경우는 L-canavanine의 세포독성이 동일하게 나타났고, 특히 caspase-8가 결손된 I9.2 세포의 경우는 L-canavanine의 세포독성에 대한 민감성이 A3와 I2.1 세포에 비해 단지 미약하게만 완화되는 것으로 나타나, L-canavanine의한 apoptosis에는 Fas/FasL 상호작용이 관련되어 있지 않으며, 또한 caspase-8의 역할이 필수적이지 않음을 시사하였다. Jurkat T 세포에 있어서 L-canavanie에 의해 유도되는 sub-$G_1$ peak 및 caspases 활성화에 미치는 pan-caspase inhibitor (z-VAD-fmk), caspase-9 inhibitor (z-LEHD-fmk), caspase-3 inhibitor (z-DEVD-fmk), caspase-4 inhibitor (z-LEVD-fmk) 및 caspase-12 inhibitor (z-ATAD-fmk)의 영향을 조사한 결과, L-canavanie에 의한 apoptosis는 ${\Delta\Psi}m$의 감소, caspase-9 및 caspase -3의 활성화에 뒤따른 caspase-8 및 caspase-7의 활성화, 그리고 PARP의 분해의 순서로 유도되는 것으로 나타났으며, 아울러 caspase-9의 활성화와 함께 caspase-12의 활성화가 L-canavanine 처리에 따른 caspase-3의 활성화에 요구되는 것으로 확인되었다. 결론적으로, L-canavanine 처리에 의한 Jurkat T 세포의 apoptosis는 ${\Delta\Psi}m$ 감소, caspase-9, caspase-3 및 caspase-7의 활성화에 의해 유도되며, 이들 apoptosis 현상들은 $p56^{lck}$에 의해 negative regulation되었다.

• Fisheries and Aquatic Sciences
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• 제9권3호
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• pp.123-128
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• 2006

An epizootic causing mortality among cultured marine finfishes occurred in 1999 in the province of Kyoungsang, Korea. The disease was characterized by the presence of enterocele, abdominal swelling, and gastroenteritis. The causative bacteria were isolated from olive flounder (Paralichthys olivaceus), black rockfish (Sebastes schlegeli), turbot (Scophthalmus maximus) and the rearing water. These bacteria showed swarming activity on agar plates and yellowish or greenish colonies on thiosulfate-citrate-bile salts-sucrose (TCBS) agar plates, but no luminescence. The pathogen was identified as Vibrio harveyi based on morphological and biochemical characteristics and the sequence of l6S rDNA. The lethal doses (LD$_{50}$) of olive flounder and black rockfish were estimated to be $1.24\times10^6-1.36\times10^8$ and $3.24\times10^5-5.8\times10^7$ CFU/fish respectively following intraperitoneal injection.

• Fisheries and Aquatic Sciences
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• 제5권4호
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• pp.258-262
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• 2002

Effects of Escherichia coli genomic DNA on the production of phagocyte activating supernatants by the head kidney leucocytes isolated from olive flounder (Paralichthys olivaceus) were investigated. Phagocyte activating activity of the supernatants was estimated by. measuring reactive oxygen species (ROS) production in target head kidney phagocytes. All supernatants from olive flounder head kidney leucocytes-stimulated with E. coli DNA induced significantly (P<0.01) higher ROS production from target pagocytes than the unstimulated control supernatant. Maximum enhancement of chemiluminescent response was observed $5.0-10.0{\mu}g\;mL^{-1}$ of bacterial DNA while the increment ability was decreased significantly (P<0.01) at the concentration of $20.0{\mu}mL^{-1}$. The results demonstrate that olive flounder head-kidney leucocytes stimulated with bacterial DNA release a soluble phagocyte activating cytokines capable of enhancing the respiratory burst activity from target phagocytes.

• Fisheries and Aquatic Sciences
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• 제18권4호
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• pp.417-420
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• 2015

We investigated the relationship between viability and IGF-1 receptor (IGF-1R) activity in D-shaped and umbo larvae of the surf clam Spisula sachalinensis after treatment with vitrification solution (VS) or freezing. In a toxicity assay, VS1, containing 5 M dimethyl sulfoxide (DMSO), was very harmful to D-shaped and umbo larvae. However, VS2, containing 5 M ethylene glycol (EG), was not harmful to either larval stage. Although VS2 had a promising toxicity test outcome, none of the larvae survived vitrification. After immersion into VSs and freezing, IGF-1R ${\beta}$-subunits were detected in all larvae; however, tyrosine phosphorylation of intracellular ${\beta}$-subunits was detected only in the control and live groups. These results suggest that activation of IGF-1R may influence surf clam larvae viability.

• Fisheries and Aquatic Sciences
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• 제7권1호
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• pp.29-33
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• 2004

The potential of phenanthrene to induce xenobiotic responses was investigated. Olive flounder (Paralichthys olivaceus) was exposed to different levels of phenanthrene $(0.5,\;1\;and\;{\mu}M)$ for 4 weeks, Gill CYP450 content and EROD (ethoxyresorufin O-deethylation) activity were found to be significant in the flounders treated with the higher concentration of phenanthrene $(>1.0\;{\mu}M)$, however, there were no significant changes in gill PROD (penthoxyresorufin O-deethylation) activity in all treated group compared to the controls. This study demonstrated that phenanthrene has potential to induce gill cytochrome P450 and EROD enzyme in olive flounder.

• Fisheries and Aquatic Sciences
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• 제8권3호
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• pp.118-121
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• 2005

Arylsulfatase cloned from a marine bacterium, Pseudoalteromonas carrageenovora, was over-expressed in Escherichia coli. Most of the recombinant arylsulfatase was found in the cell lysate with induction up to $10{\mu}M$ IPTG. However, enzyme activity was observed both in the culture supernatant and cell lysate by induction with IPTG concentration of $50-5,000{\mu}M$. Most of the recombinant enzyme was localized in the periplasmic space with $10{\mu}M$ IPTG induction, while half of the enzyme was distributed in the periplasmic space with $50{\mu}M$ IPTG induction. Cell growth and arylsulfatase activity did not change with the induction time, and the level of recombinant arylsulfatase expression was maintained at 4-5 U/mL after 6 to 14 hr of culture.

• Fisheries and Aquatic Sciences
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• 제15권3호
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• pp.259-263
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• 2012

To isolate an alginate-degrading bacterium, we conducted a single colony isolation using a solid medium containing alginate as the sole carbon source. A marine bacterium Y-4 capable of degrading alginate was isolated from seawater. The strain was identified to be Pseudoalteromonas sp., based on morphological, biochemical, 16S rDNA homology, and phylogenetic analyses. Moreover, Pseudoalteromonas sp. Y-4 exhibited alginate lyase activity in the presence of 4% alginate even though many known alginate-degrading bacteria degrade in the range of 0.5-1% alginate. The optimum culture conditions for the Y-4 strain were 2% alginate, pH 8.0, and 3% NaCl at $30^{\circ}C$. The highest alginate lyase activity was also observed under the same conditions. To our knowledge, this is the first reported isolation of a marine bacterium degrading high concentrations of alginate.

• Fisheries and Aquatic Sciences
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• 제13권1호
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• pp.12-17
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• 2010

This study was conducted to evaluate the potential of a microbial fermentation procedure to improve the functional qualities of seaweeds. Aspergillus oryzae, which has been used in traditional Korean fermented foods, was inoculated and cultivated in an aqueous extract of sea tangle (Laminaria japonica). Fermentation of the sea tangle extract by A. oryzae for 4 days resulted in a 3-fold increase in $\gamma$-aminobutyric acid (GABA) content. GABA is known to be a bioactive compound. Fungal fermentation of the extract also enhanced its antioxidant activity and increased its total content of phenolic compounds. It was assumed that these changes stemmed from the biodegradation of active compounds of the sea tangle packaged within its rigid structural matrix or occurred as result of fungal fermentation. These results suggested that the application of microbial fermentation to the processing of seaweeds will help in the development of processed foods to meet consumer demands.