• Title/Summary/Keyword: bioactivity analysis

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Surface characteristics and stability of implants treated with alkali and heat (알칼리와 열처리에 의한 임플란트의 표면 특성 및 골유착 안정성에 관한 연구)

  • Song, Yun-Seok;Cho, In-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.5
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    • pp.490-499
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    • 2008
  • Statement of problem: Bioactive materials must have the ability to spontaneously form a bone like apatite layer on their surface and induce direct biochemical bonding to bone. A simple chemical treatment via alkali and heat has been revealed to induce bioactivity in titanium. Purpose: The purpose of this study was to evaluate the surface characteristics and stability of alkali and heat treated implants. Material and methods: Specimens were divided into three groups; group 1 was the control group with machined surface implants, groups 2 and 3 were treated with alkali solutions and heat treated in the atmosphere and vacuum conditions respectively. The surface characteristics were observed with FESEM, XPS, TF-XRD and AFM. Stability was evaluated with the resonance frequency analysis, periotest and removal torque values. One-way ANOVA and Duncan test were used for statistical analysis. Results: 1. Groups treated with alkali and heat showed similar characteristics. Groups 2 and 3 showed high compositions of Na ions on the surface with sub-micron sized pores compared to group 1. Group 2 showed mixed compositions of anatase and rutile with superior contents of rutile. 2. Resonance frequency analysis : The ISQ of group 2 showed significantly higher values than that of groups 1 and 3 at 12 weeks. The ISQ of groups 1 and 2 showed significant increase after 4 weeks, and the ISQ of group 3 increased significantly after 2 and 4 weeks respectively (P < .05). 3. Periotest: The PTV of groups 1 and 2 showed significant decrease after 4 weeks, and the PTV of group 3 showed significant decrease after 2 and 4 weeks respectively (P < .05). 4. Removal torque analysis: The removal torque value of group 2 was significantly higher than those of groups 1 and 3 at 2, 4 and 8 weeks. The removal torque values of groups 1 and 3 showed increase at 4 and 12 weeks, but the removal torque value of group 2 showed increase after 4 weeks (P < .05). Conclusion: An oxide layer with appropriate crystal structure and amorphous sodium titanate layer can be obtained on titanium implants through alkali and heat treatment in the atmosphere, and even alkali and heat treatment in vacuum conditions, provided a bioactive surface containing sodium. These surface layers can be considered to be effective for enhancement of osseointegration and reduction of healing period for implant treatment.

Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

Evaluation of Commercial Korean Honey Quality and Correlation Analysis of the Quality Parameters (국내 시판 벌꿀의 품질 평가 및 품질인자간 상관관계 분석)

  • Sung, Hwa-Jung;Jung, Chuleui;Kwon, Jiyoung;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1489-1500
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    • 2018
  • Honey is made from flower nectar by honey bees. In this study, 120 honeys from various flowers and across eight different provinces in Korea were collected and their components, antioxidants, and hemolytic activities against red blood cell were evaluated. Our results show that total polyphenol (TP) varied widely across the samples, with chestnut honey showing the highest TP ($77.1{\pm}8.4mg/100g$), protein content ($25.9{\pm}0.9mg/100g$), and absorbance at 400 nm ($A_{400}$ : $0.156{\pm}0.036$). In contrast, the acacia honey and sugar honey had a TP of 9.5~30 mg, 12~15 mg/100g of, and the lowest $A_{400}$ of $0.06{\pm}0.02$. High amounts of total flavonoid were quantified in the jujube and chestnut honeys at $8.73{\pm}7.31$ and $8.39{\pm}3.02mg/100g$, respectively. No samples demonstrated hemolytic activity up to 1 mg/ml. Antioxidant activities determined by DPPH, ABTS, and nitrite scavenging placed the chestnut honey highest, followed by jujube, styrax, multi-floral, citrus, acacia and sugar honey. Analysis of parameter correlations indicated that the components and bioactivity of the honey are dependent on the origin of the flower rather than on bee-farming regions. A positive correlation between TP content and antioxidant activity was identified. The correlation coefficients between $A_{400}$ and the TP, ABTS scavenging, and reducing power values were 0.804, 0.772 and 0.741, respectively. We therefore suggest that $A_{400}$ could be used as a noble, economic and simple factor for honey quality evaluation. Our results can potentially be used to develop functional honey for the food and pharmaceutical industries.

Effect of Scytosiphon lomentaria Ethanol Extracts on Myostatin Activity and Zebrafish Obesity Induced by High Feeding (고리매(Scytosiphon lomentaria) 에탄올 추출물이 마이오스타틴 활성과 고 급식으로 유도된 비만 제브라피쉬에 미치는 영향)

  • Jung, Jun Gyo;Kim, Jae Hong;Kim, Jeong Hwan;Kim, Yong Soo;Jin, Deuk-Hee;Jin, Hyung-Joo
    • Journal of Life Science
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    • v.31 no.8
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    • pp.699-709
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    • 2021
  • Muscle mass improvement through lifestyle modification has been shown to reduce the risk of metabolic syndrome. This study examined the capacity of ethanol extracts of Scytosiphon lomentaria (SLE) to suppress the bioactivity of myostatin, a potent negative regulator of skeletal muscle mass, as well as the effect of SLE treatment on metabolic homeostasis in obese zebrafish induced by high feeding. A total of 10 ㎍/ml SLE completely blocked myostatin (1 nM/ml) signaling in the pGL3-(CAGA)12 luciferase assay and suppressed myostatin-induced Smad2 phosphorylation in the Western blot analysis. In the zebrafish larvae analysis, the whole body glucose concentration of the high feeding control (HFC) group was significantly higher than that of the normal feeding control (NFC) group. However, the glucose levels of the high feeding group treated with 12.5 ug SLE and of the high feeding group treated with 18.75 ug SLE were similar to those of the NFC group. The mRNA expression level of the GLUT2 gene of the HFC group was significantly lower than that of the NFC group. SLE treatment restored the expression of the GLUT2 gene to a level that was close to that of the NFC group, indicating that SLE is capable of regulating glucose levels in zebrafish larvae. The current results highlight the potential of SLE as a natural MSTN inhibitor and supplement that can be used to facilitate the treatment of metabolic syndrome.

Analysis of Terpenoids as Volatile Compound Released During the Drying Process of Cryptomeria japonica (삼나무 건조 중 발생하는 휘발성 유기화합물 Terpenoids의 분석)

  • Lee, Su-Yeon;Gwak, Ki-Seob;Kim, Seon-Hong;Lee, Jun-Jae;Yeo, Hwan-Myeong;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.3
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    • pp.242-250
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    • 2010
  • The aim of this study was to investigate the terpenoids of Total Volatile Organic Compounds (VOCs) released during drying of Cryptomeria japonica using the thermal extractor (TE). Considering the drying process of C. japonica, temperatures of TE were set at $27^{\circ}C$, $60^{\circ}C$, $80^{\circ}C$, $100^{\circ}C$, and $120^{\circ}C$, respectively. As the result, the emission factors of VOCs and terpenoids were increased as temperature increased. The amount of terpenoids included in VOCs emission factors were 87.5%, 81.6%, 83.6%, 90.1%, and 97.3% depending on above temperatures, respectively. Especially at$100^{\circ}C$ and $120^{\circ}C$, the amount of terpenoids were measured more than 90%. ${\delta}$-cadinene was the highest yield at each temperature and 32 types of terpenoids were collected. Emitted terpenoids were classified into the sesquiterpene group which consists of 15 carbon sources. These 32 sesquiterpenes were used for determining the useful bioactivity such as antifungal activity by the agar dilution. As the result, they showed the antifungal activity against Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum gypseum. The 5,000 ppm concentration of terpenoids showed a strong activity with 100% against the 3 fungi. At the 1,000 ppm concentration of terpenoids, the antifungal activities against three fungi were 95.2%, 98.7%, and 97.3%, and their activities were a little inhibited at 100 ppm concentration.

Effect of Alkaline Ionized Water on Stabilization of Antioxidation, Antithrombosis and Antibacterial Activities (항산화, 항혈전 및 항세균 활성의 안정화에 미치는 알칼리 이온수의 영향)

  • Ahn, Seon-Mi;Kang, Mee-A;Kim, Moo-In;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1107-1112
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    • 2010
  • In this study, the stable maintenance of bioactivity in alkaline ionized water (AIW) and antibacterial effects of AIW were evaluated to confirm benefits of AIW. As controls, purified water (PW) and tap drinking water (DW) were used. The pH and ORP (oxidation-reduction potential) of AIW, PW and DW used were 9.5 and 120 mV, 7.2 and 144 mV, and 7.3 and 564 mV, respectively. High level of minerals was observed in DW (DW>AIW>PW of mineral contents). Concentrations of $Ca^{++}$ and $Na^+$ in DW were 14.5, and 8.4 mg/l, respectively, while no $Ca^{++}$, $Mg^{++}$, $K^+$, and $Na^+$ were detected in PW. Evaluation of antioxidant activities for AIW, PW and DW showed that the waters did not act as antioxidants. However, the DPPH (1,1-diphenyl-2-picryl hydrazyl) or superoxide radical scavenging activities or reducing power of vitamin C were stably maintained in AIW and PW, though not in DW, against heat treatment ($60^{\circ}C$) or vigorous shaking (120 rpm) at $37^{\circ}C$. Similarly, after aspirin treatment at $60^{\circ}C$ for 1 hr, the antithrombosis activity in PW and AIW was 62.6% and 55.3%, while that of DW was 52.1%. Furthermore, cell growth analysis and viable cell count of Escherichia coli H7:O157 in PW, AIW and DW showed that AIW and DW, not DW, have antibacterial activities. Our results suggest that the state of water, for example pH, ORP and mineral contents of water, should be considered in medicine or food industries, and that AIW has high potential for utilization in various fields.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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