• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1881-1890
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• 2016

Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

• Korean Journal of Pharmacognosy
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• v.40 no.2
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• pp.150-154
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• 2009

We examined ethanol extracts prepared from the Korean marine algae belonging to the Sargassaceae family for their inhibitory effects on ${\alpha}$-glucosidase activity and free radicals in vitro. Among five marine algae, the extracts of Sargassum yezoense were found to possess strongly ${\alpha}$-glucosidase inhibition and free radicals scavenging activities. Two compounds were isolated via bioactivity guided isolation and tested for their effects on ${\alpha}$-glucosidase, DPPH, $ABTS^{+}$ and $Photochem^{(R)}$ analysis. Their chemical structures were elucidated by spectral analysis and direct comparison with authentic compounds; their structures were identified as sargaquinoic acid (1) and sargahydroquinoic acid (2). The inhibitory effects of compound 1 and 2 ($IC_{50}$ value:14.2 and 12.8 ${\mu}M$, respectively) on ${\alpha}$-glucosidase were more potent that of deoxynojirimycin as a positive control ($IC_{50}$ value:18.0 ${\mu}M$). All compounds displayed antioxidative activity which was measured by DPPH, $ABTS^{+}$ and $Photochem^{(R)}$ apparatus.

• Polymer(Korea)
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• v.39 no.3
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• pp.493-498
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• 2015

For biomaterials for skin regeneration with minimized inflammatory response, high bioactivity and biocompatibility are highly required. Also, it should have a porous microstructure to improve cell adhesion and growth. In this study, we extracted a new collagen source from duck's feet which is by-product, and made the shape of sponges from duck's feet collagen (DC) to compare with DBP and SIS. To analyze physical and chemical property of the scaffold, SEM and FTIR were used. MTT assay was used to measure the attachment and proliferation of NIH/3T3 in the scaffolds. RTPCR was used to evaluate the expression of proinflammatory cytokine. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) was used to measure the ability of antioxidant activity. Overall, this study shows that DC scaffold is biocompatible and has good physical property. Additionally, DC scaffold shows the potential as wound healing biomaterials.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.280-286
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• 2009

To investigate anti-phytopathogenic fungal activity of Coptis chinensis, the methanol extract and its organic solvent fractions were prepared. Using the extract and the fractions, in-vitro spore-germination inhibition and mycelial-growth inhibition activities were evaluated against Colletotrichum gloeosporioides, Phytohpthora capsici, Pyricularia grisea, Rhizoctonia solani, Botryosphaeri dothidea, Glomerella cingulata, respectively. Treatment of the methanol extract (500 mg/mL) into the spore of phytopathogenic fungi completely inhibited germinations for 5 days, except B. dothidea, and showed strong antifungal activities against P. grisea and B. cinerea, and antioomycetes activity against P. capsici. The minimal growth inhibition concentrations of the methanol extract against P. grisea, B. cinerea and P. capsici were 300, 300, and 500 mg/mL, respectively. For practical application of C. chinensis in red-pepper field, the hot-water extract (1,000 mg/mL) was prepared in commercial facility, after evaluation of heat stability and solvent-extraction yields of antifungal substances. The 3-times leaf-spray of the extract from June to August, 2008 did not show any deleterious effect to red-pepper. In fact, the leaf-spray promoted plant growth including leaf, root and fruit. The average weight and rind of each fruit were increased to 119% and 117% comparison to those of without treatments. Our results suggest that C. chinensis is a useful source for control of red-pepper diseases and plant growth.

• Molecular & Cellular Toxicology
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• v.1 no.3
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• pp.172-178
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• 2005

[ $21{\alpha}$ ]- and ${\beta}$-Methylmelianodiol were isolated as the inhibitor of IL-5 bioactivity from Poncirus tripoliata. To develope as an anti-septic drug, the genotoxicity of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ was subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. These compounds are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. Before performing the comet assay, $IC_{20}$ of $21{\alpha}-methylmelianodiol$ was determined the concentration of $25.51\;{\mu}g/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. Also $21{\beta}-methylmelianodiol$ was determined the concentration of $24.15\;{\mu}g/mL\;and\;\;22.46\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, DNA damage was not observed both $21{\alpha}-methylmelianodiol\;and\;21{\beta}-methylmelianodiol$ in mouse lymphoma cell line. Also, the mutant frequencies in the treated cultures were similar to the vehicle controls, and none of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ with and without S-9 doses induced a mutant frequency over. twice the background. It is suggests that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ are non-mutagenic in MOLY assay. The results of this battery of assays indicate that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ have no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$, as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.57 no.1
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• pp.1-14
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• 2024

We investigated the functional properties and in vitro bioactivity of protein isolates (RPIs) recovered from olive flounder Paralichthys olivaceus roes by isoelectric solubilization/precipitation process, according to pH-shift treatments. The buffer capacity of RPIs was shown to be stronger in alkaline pH than in acidic pH. Water holding capacity of RPIs was in range of 4.5-5.2 g/g protein with no significant differences (P>0.05). The foaming capacity and emulsifying activity index of RPIs did not show any significant differences between RPI-1 (pH 11/4.5) and 3 (pH 12/4.5), however they were superior to RPI-2 (pH 11/5.5) and 4 (pH 12/5.5). The 2,2'-azino-bis-3-ethylbenzo-thiazoline-6-sulfonic acid radical scavenging activity of RPI-3 (2.5 mg protein/mL) was 102.4 ㎍/mL, and the angiotensin I converting enzyme inhibitory activity was 30.8%. Among the RPIs, RPI-3 was relatively superior in protein functional properties such as buffer capacity, foaming capacity, emulsification, and anti-oxidative activity. Therefore, we suggest that RPI prepared from olive flounder roes could serve as a potential food resource.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.5
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• pp.434-441
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• 2009

This study was conducted to compare the biological activities of 7 melania snails from the family Pleuroceridae (Semisulcospira coreana, Koreanomelania nodifila, Semisulcospira forticosta, Koreoleptoxis globus ovalis, Semisulcospira libertina, Semisulcospira tegulata and Semisulcospira gottschei) in Korea. Among the 7 species, S. coreana, Korean. nodifila, S. forticosta and S. gottschei showed over 80% cytotoxicities on three cancer cell lines (SNU-1, A549 and Hep 3B) compared to the non-treatment, whereas S. libertina and S. tegulata showed almost no growth inhibition activities on the same cancer cell lines. In relation to ACE inhibition activity, only S. coreana, Korean. nodifila, and S. forticosta showed over 60% ACE inhibition activities, whereas other melania snails exhibited inhibition activities of lower than 25%. DPPH radical scavenging activities were also determined, and used to categories melania snails into three groups based on Duncan's multiple range test at P<0.05. The amount of TNF-${\alpha}$ produced by in vitro mouse peritoneal macrophage was determined according to bioactivity on L-929 cells. Three melania snails, S. coreana, Korean. nodifila and S. gottschei, exhibited 95.2%, 89.7% and 93.7% cell death(%) on L-929 cells, respectively. Glucose-6-phosphate dehydrogenase inhibitory activity was also obtained in the extract of S. coreana (31.9%) and Korean. nodifila (28.1%), showing that these extracts can be used as supplemental dietary health foods. In conclusion, we believe that the extracts of melania snails should be given due consideration in functional health food development.

• Journal of Life Science
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• v.23 no.2
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• pp.249-258
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• 2013

The antioxidative activity and bioactivity of water, ethanol, and methanol extracts from Paecilomyces japonica (PJ), Cordyceps militaris (CM), and cordycepin-enriched C. militaris JLM0636 ($CM{\alpha}$) were tested in in vitro experimental models. The PJ water extract showed the highest extraction yield (42.53%). The highest content of phenolic compounds and flavonoids were found in the water extract of PJ, 2.72% and 1.73%, respectively. The major minerals were K, Mg, and Ca. The water extracts of PJ also showed the highest DPPH free radical scavenging activity and reducing power. Linoleic acid peroxidation and antioxidative activities were strong in $CM{\alpha}$. The methanol extracts of PJ showed the highest inhibition activity against tyrosinase. Fibriolytic activity was higher in $CM{\alpha}$ than in CM. These results may provide basic data to understand the biological activities of bioactive materials derived from $CM{\alpha}$ for the development of functional foods, cosmetics, and antithrombotics.

• The Journal of Advanced Prosthodontics
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• v.5 no.4
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• pp.402-408
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• 2013

PURPOSE. The aim of this study was to evaluate the surface properties and in vitro bioactivity to osteoblasts of magnesium and magnesium-hydroxyapatite coated titanium. MATERIALS AND METHODS. Themagnesium (Mg) and magnesium-hydroxyapatite (Mg-HA) coatings on titanium (Ti) substrates were prepared by radio frequency (RF) and direct current (DC) magnetron sputtering.The samples were divided into non-coated smooth Ti (Ti-S group), Mg coatinggroup (Ti-Mg group), and Mg-HA coating group (Ti-MgHA group).The surface properties were evaluated using scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The surface roughness was evaluated by atomic force microscopy (AFM). Cell adhesion, cell proliferation and alkaline phosphatase (ALP) activity were evaluated using MC3T3-E1 cells. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed. RESULTS. Cross-sectional SEM images showed that Mg and Mg-HA depositionson titanium substrates were performed successfully. The surface roughness appeared to be similaramong the three groups. Ti-MgHA and Ti-Mg group had improved cellular responses with regard to the proliferation, alkaline phosphatase (ALP) activity, and bone-associated markers, such as bone sialoprotein (BSP) and osteocalcin (OCN) mRNA compared to those of Ti-S group. However, the differences between Ti-Mg group and Ti-MgHA group were not significant, in spite of the tendency of higher proliferation, ALP activity and BSP expression in Ti-MgHA group. CONCLUSION. Mg and Mg-HAcoatings could stimulate the differentiation into osteoblastic MC3T3-E1 cells, potentially contributing to rapid osseointegration.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3519-3528
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• 2012

Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.