• Title/Summary/Keyword: biomolecule

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Fabrication of Au Nanoparticle for Au-conjugate Immuno Chemistry Probe (Au-conjugate 면역화학 진단용 금 나노입자 제조)

  • Park, Sung-Tae;Lee, Kwang-Min
    • Korean Journal of Materials Research
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    • v.13 no.8
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    • pp.550-554
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    • 2003
  • Current nanogold cluster synthesized by chemical routine with 11 or 55 atoms of gold has been widely used for immuno chemistry probe as a form of nanocluster conjugated with biomolecules. It would be an undeveloped region that the 1 nm size of nanogold could be made by materials engineering processing. Therefore, objective of this study is to minimize the size of gold nanocluster as a function of operating temperature and chamber pressure in inert gas condensation (IGC) processing. Evaporation temperature was controlled by input current from 50 A to 65 A. Chamber pressure was controlled by argon gas with a range of 0.05 to 2 torr. The gold nanocluster by IGC was evaluated by X-ray diffraction (XRD) and transmission electron microscopy (TEM). The gold nanocluster for TEM analysis was directly sampled with special in-situ method during the processing. Atomic force microscopy (AFM) was used to observe 3-D nanogold layer surfaces on a slide glass for the following biomolecule conjugation step. The size of gold nanoclusters had a close relationship with the processing condition such as evaporation temperature and chamber pressure. The approximately 1 nm size of nanogold was obtained at the processing condition for 1 torr at $1124 ^{\circ}C$.

Interaction at the nanoscale of fundamental biological molecules with minerals

  • Valdre, Giovanni;Moro, Daniele;Ulian, Gianfranco
    • Advances in nano research
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    • v.1 no.3
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    • pp.133-151
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    • 2013
  • The availability of advanced nanotechnological methodologies (experimental and theoretical) has widened the investigation of biological/organic matter in interaction with substrates. Minerals are good candidates as substrates because they may present a wide variety of physico-chemical properties and surface nanostructures that can be used to actively condense and manipulate the biomolecules. Scanning Probe Microscopy (SPM) is one of the best suited techniques used to investigate at a single molecule level the surface interactions. In addition, the recent availability of high performance computing has increased the possibility to study quantum mechanically the interaction phenomena extending the number of atoms involved in the simulation. In the present paper, firstly we will briefly introduce new SPM technological developments and applications to investigate mineral surfaces and mineral-biomolecule interaction, then we will present results on the specific RNA-mineral interaction and recent basics and applicative achievements in the field of the interactions between other fundamental biological molecules and mineral surfaces from both an experimental and theoretical point of view.

The Study of DNA Damage Induced by Atmospheric Pressure Plasma Jet and Their Mechanisms

  • Park, Yeunsoo;Song, Mi-Young;Yoon, Jung-Sik
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.155.1-155.1
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    • 2013
  • The goals of this study are to elucidate the plasma effects on DNA molecules to apply some plasma based applications and also to find out the mechanisms of plasma-induced DNA damage in biomolecule. Nonthermal atmospheric pressure plasma has much potential for medical, agricultural and food applications for the future. The atmospheric pressure plasma jet (APPJ) contains radicals, charged particles, low energy electrons, excited molecules and UV light. It has been started doing experiments using APPJ at the early 21th. And some recent results showed that APPJ has a possibility to apply to new fields like mentioned above. But it is kind of at the very early stages of plasma based application. It is definitely necessary much of theoretical and experimental studies to further understanding to use nonthermal atmospheric pressure plasma in biomedical, agriculture and food parts. Here we introduce a new experimental system to study plasma effects on biomolecules. And we will show some recent results of LEE-induced DNA damage using electron irradiation apparatus under ultra-high vacuum.

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The Effect of Various Methods of Cross-linking in Type I Collagen Scaffold on Cartilage Regeneration (I형 콜라겐지지체의 다양한 가교처리 방법이 연골막성 연골재생에 미치는 영향)

  • Son, Dae Gu;Lim, Joong Jae;Sohn, Kyounghee;Yang, Eun Kyung
    • Archives of Plastic Surgery
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    • v.33 no.6
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    • pp.723-731
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    • 2006
  • Purpose: Collagen is the principal structural biomolecule in cartilage extracellular matrix, which makes it a logical target for cartilage engineering. In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four types of scaffolds(n=4) were inserted. The types included DHT cross-linked(Group 1), DHT and EDC cross- linked(Group 2), CS added DHT cross-linked(Group 3), and CS added DHT and EDC cross-linked(Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated respectively 4, 8, and 12 weeks after implantation. Results: The largest quantity of regenerated cartilage was found in DHT cross-linked groups 1 and 3 in the 8th week and then decreased in the 12th week, while calcification increased. Calcification was observed from the 8th week and the area increased in the 12th week. Group 4 was treated with EDC cross-linking and CS, and the matrix did not degrade in the 12th week. Cartilage-specific type II collagen mRNA expression increased with time in all groups. Conclusion: CS did not increase chondrogenesis in all groups. EDC cross-linking may prevent chondrocyte infiltration from the perichondrium into the collagen scaffold.

Transcription Analysis of Daptomyc in Biosynthetic Genesin Streptomyces roseosporus

  • Rhee, Ki-Hyeong;Davies, Julian
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1841-1848
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    • 2006
  • Insights into gene expression have the potential for improvement of antibiotic yield and the development of robust production hosts for use in recombinant biomolecule production. $Cubicin^{TM}$ (daptomycin for injection) is a recently approved antibiotic active against many Gram(+) pathogens, including those resistant to methicillin, vancomycin, and fluoroquinolones. Daptomycin is produced as a secondary metabolite by Streptomyces roseosporus. A 128 kb region of DNA including the daptomycin biosynthetic gene cluster (dpt) has been cloned. and sequenced. Using a selected array of nucleic acid probes representing this region, we compared the expression levels of the dpt genes between S. roseosporus wild-type (WT) and derived S. roseosporus high-producer of daptomycin (HP). We observed that the majority of the biosynthetic genes were upregulated in HP compared with WT; a total of 12 genes, including those encoding daptomycin synthetase, showed consistently and significantly higher expression levels, at least 5-fold, in HP compared with WT. In contrast, some genes, flanking the dpt cluster, were expressed at higher levels in the WT strain. The expression of housekeeping genes such as S. roseosporus rpsL, rpsG, and 16S (positive controls) and presumptive intergenic regions in the dpt cluster (negative control) were identical in the two strains. In addition, we compared transcription during the early, mid-log, and early-stationary phases of growth in the HP strain. The same set of genes was upregulated and downregulated under all conditions examined; housekeeping genes showed no relative change in expression level over the periods of growth tested. Analyses of this type would be of value in studies of strain improvement and also for the identification of gene regulation processes that are important for secondary metabolite production.

S-Adenosylmethionine (SAM) Regulates Antibiotic Biosynthesis in Streptomyces spp. in a Mode Independent of Its Role as a Methyl Donor

  • Zhao Xin-Qing;Jin Ying-Yu;Kwon Hyung-Jin;Yang Young-Yell;Suh Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.927-932
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    • 2006
  • S-Adenosylmethionine (SAM) is a ubiquitous biomolecule serving mainly as a methyl donor. Our recent studies revealed that SAM controls antibiotic production in Streptomyces. In this study, the functional mode of SAM was studied in S. coelicolor and S. antibioticus ATCC11891, employing S-adenosylhomocysteine (SAH), a methylation reaction product of SAM. Actinorhodin biosynthesis did not require SAM as a methyl donor, whereas SAH enhanced the actinorhodin biosynthesis up to the level comparable to SAM, and the most effective concentration of SAH was higher than that of SAM. In the case of oleandomycin that requires SAM for its biosynthesis, both SAM and SAH at the concentration as low as 100 mM showed comparable efficacy in enhancing the production; SAM at 1 mM concentration additionally stimulated to give a 5-fold enhancement of oleandomycin production. In vitro autophosphorylation of protein kinase AfsK was found to be activated by both SAM and SAH, as well as other structurally related compounds. Our studies demonstrate that SAM regulates antibiotic biosynthesis in a mode independent of its role as a methyl donor and suggest that SAM acts directly as an intracellular signaling molecule for Streptomyces.

Benzo(a)pyrene-Triglyceride Adduct: a Potential Molecular Biomarker for Carcinogen Exposure

  • Lee, Byung-Mu
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05b
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    • pp.20-45
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    • 2002
  • In this study, we demonstrated the in vitro and in vivo formation of carcinogen-lipid adduct and its correlation with DNA or protein adducts. The lipids from serum or hepatocyte membranes of Spragu-Dawley rats. human serum, and standard major lipids were in vitro reacted with benzo[a]pyrene(BP) and BP metabolites. 7,8-Dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]-pyrene(BPDE-I), an ultimate carcinogenic form of BP, was covalently bound to triglyceride(TG). BPDE-I-TG adducts isolated by thin-layer chromatography (TLC) were further detected by high performance liquid chromatography(HPLC). TGs, including triolein, tripalmitin and tristearin, showed positive reactions with BPDE-I. However, cholesterol, phospholipids(Phosphatidylcholine, phosphatidyl-ethanolamine, phosphatidyl-inositol and sphingomyelin) and nonesterified fatty acids(palmitic acid, oleic acid, linoleic acid and stearic acid) did not react with BPDE-I. In addition, other BP metabolites (BP-phenols and -diols) did not react with TG, which TG appeared to be the most reactive lipid yet studied with respect to its ability to form an adduct with BPDE-I. There was a clear-cut dose-respect to its ability to form an adduct with BPDE-I-lipid adduct in vitro between TG and [1,3-3H]BPDE-I. In an animal study, BPDE-I-TG was also formed in the serum of rats orally treated with BP(25 mg/rat). Also, obvious correlations between [3H]BP related-biomolecule adducts (DNA, protein) or lipid damage and the BPDE-I-TG adduct were obtained in various tissues of mice i.p. treated with [3H]BP. These data suggest that TG can form an adduct with BPDE-I, as do other macromolecules (DNA, RNA, and protein). Therefore, a carcinogen-lipid adduct would be a useful biomarker for chemical carcinogenesis research and cancer risk assessment.

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Reduced graphene oxide field-effect transistor for biomolecule detection and study of sensing mechanism

  • Kim, D.J.;Sohn, I.Y.;Kim, D.I.;Yoon, O.J.;Yang, C.W.;Lee, N.E.;Park, J.S.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.431-431
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    • 2011
  • Graphene, two dimensional sheet of sp2-hybridized carbon, has attracted an enormous amount of interest due to excellent electrical, chemical and mechanical properties for the application of transparent conducting films, clean energy devices, field-effect transistors, optoelectronic devices and chemical sensors. Especially, graphene is promising candidate to detect the gas molecules and biomolecules due to the large specific surface area and signal-to-noise ratios. Despite of importance to the disease diagnosis, there are a few reports to demonstrate the graphene- and rGO-FET for biological sensors and the sensing mechanism are not fully understood. Here we describe scalable and facile fabrication of rGO-FET with the capability of label-free, ultrasensitive electrical detection of a cancer biomarker, prostate specific antigen/${\alpha}1$-antichymotrypsin (PSA-ACT) complex, in which the ultrathin rGO sensing channel was simply formed by a uniform self-assembly of two-dimensional rGO nanosheets on aminated pattern generated by inkjet printing. Sensing characteristics of rGO-FET immunosensor showed the highly precise, reliable, and linear shift in the Dirac point with the analyte concentration of PSA-ACT complex and extremely low detection limit as low as 1 fg/ml. We further analyzed the charge doping mechanism, which is the change in the charge carrier in the rGO channel varying by the concentration of biomolecules. Amenability of solution-based scalable fabrication and extremely high performance may enable rGO-FET device as a versatile multiplexed diagnostic biosensor for disease biomarkers.

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Biodistribution of 99mTc Tricarbonyl Glycine Oligomers

  • Jang, Beom-Su;Lee, Joo-Sang;Rho, Jong Kook;Park, Sang Hyun
    • Toxicological Research
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    • v.28 no.4
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    • pp.235-240
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    • 2012
  • $^{99m}Tc$ tricarbonyl glycine monomers, trimers, and pentamers were synthesized and evaluated for their radiolabeling and in vivo distribution characteristics. We synthesized a $^{99m}Tc$-tricarbonyl precursor with a low oxidation state (I). $^{99m}Tc(CO)_3(H_2O)_3^+$ was then made to react with monomeric and oligomeric glycine for the development of bifunctional chelating sequences for biomolecules. Labeling yields of $^{99m}Tc$-tricarbonyl glycine monomers and oligomers were checked by high-performance liquid chromatography. The labeling yields of $^{99m}Tc$-tricarbonyl glycine and glycine oligomers were more than 95%. We evaluated the characteristics of $^{99m}Tc$-tricarbonyl glycine oligomers by carrying out a lipophilicity test and an imaging study. The octanol-water partition coefficient of $^{99m}Tc$ tricarbonyl glycine oligomers indicated hydrophilic properties. Single-photon emission computed tomography imaging of $^{99m}Tc$-tricarbonyl glycine oligomers showed rapid renal excretion through the kidneys with a low uptake in the liver, especially of $^{99m}Tc$ tricarbonyl triglycine. Furthermore, we verified that the addition of triglycine to prototype biomolecules (AGRGDS and RRPYIL) results in the improvement of radiolabeling yield. From these results, we conclude that triglycine has good characteristics for use as a bifunctional chelating sequence for a $^{99m}Tc$-tricarbonyl-based biomolecular imaging probe.

Novel Antibacterial, Cytotoxic and Catalytic Activities of Silver Nanoparticles Synthesized from Acidophilic Actinobacterial SL19 with Evidence for Protein as Coating Biomolecule

  • Wypij, Magdalena;Ostrowski, Maciej;Piska, Kamil;Wojcik-Pszczola, Katarzyna;Pekala, Elzbieta;Rai, Mahendra;Golinska, Patrycja
    • Journal of Microbiology and Biotechnology
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    • v.32 no.9
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    • pp.1195-1208
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    • 2022
  • Silver nanoparticles (AgNPs) have potential applications in medicine, photocatalysis, agriculture, and cosmetic fields due to their unique physicochemical properties and strong antimicrobial activity. Here, AgNPs were synthesized using actinobacterial SL19 strain, isolated from acidic forest soil in Poland, and confirmed by UV-vis and FTIR spectroscopy, TEM, and zeta potential analysis. The AgNPs were polydispersed, stable, spherical, and small, with an average size of 23 nm. The FTIR study revealed the presence of bonds characteristic of proteins that cover nanoparticles. These proteins were then studied by using liquid chromatography with tandem mass spectrometry (LC-MS/MS) and identified with the highest similarity to hypothetical protein and porin with molecular masses equal to 41 and 38 kDa, respectively. Our AgNPs exhibited remarkable antibacterial activity against Escherichia coli and Pseudomonas aeruginosa. The combined, synergistic action of these synthesized AgNPs with commercial antibiotics (ampicillin, kanamycin, streptomycin, and tetracycline) enabled dose reductions in both components and increased their antimicrobial efficacy, especially in the case of streptomycin and tetracycline. Furthermore, the in vitro activity of the AgNPs on human cancer cell lines (MCF-7, A375, A549, and HepG2) showed cancer-specific sensitivity, while the genotoxic activity was evaluated by Ames assay, which revealed a lack of mutagenicity on the part of nanoparticles in Salmonella Typhimurium TA98 strain. We also studied the impact of the AgNPs on the catalytic and photocatalytic degradation of methyl orange (MO). The decomposition of MO was observed by a decrease in intensity of absorbance within time. The results of our study proved the easy, fast, and efficient synthesis of AgNPs using acidophilic actinomycete SL19 strain and demonstrated the remarkable potential of these AgNPs as anticancer and antibacterial agents. However, the properties and activity of such particles can vary by biosynthesized batch.