Search results for: cationic peptide
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 349

Search results for: cationic peptide

139 Analysis of Osmotin as Transcription Factor/Cell Signaling Modulator Using Bioinformatic Tools

Authors: Usha Kiran, M. Z. Abdin

Abstract:

Osmotin is an abundant cationic multifunctional protein discovered in cells of tobacco (Nicotiana tabacum L. var Wisconsin 38) adapted to an environment of low osmotic potential. It provides plants protection from pathogens, hence placed in the PRP family of proteins. The osmotin induced proline accumulation has been reported in plants including transgenic tomato and strawberry conferring tolerance against both biotic and abiotic stresses. The exact mechanism of induction of proline by osmotin is however, not known till date. These observations have led us to hypothesize that osmotin induced proline accumulation could be due to its involvement as transcription factor and/or cell signal pathway modulator in proline biosynthesis. The present investigation was therefore, undertaken to analyze the osmotin protein as transcription factor /cell signalling modulator using bioinformatics tools. The results of available online DNA binding motif search programs revealed that osmotin does not contain DNA-binding motifs. The alignment results of osmotin protein with the protein sequence from DATF showed the homology in the range of 0-20%, suggesting that it might not contain a DNA binding motif. Further to find unique DNA-binding domain, the superimposition of osmotin 3D structure on modeled Arabidopsis transcription factors using Chimera also suggested absence of the same. We, however, found evidence implicating osmotin in cell signaling. With these results, we concluded that osmotin is not a transcription factor but regulating proline biosynthesis and accumulation through cell signaling during abiotic stresses.

Keywords: osmotin, cell signaling modulator, bioinformatic tools, protein

Procedia PDF Downloads 239
138 In Vitro Antioxidant Properties of Balanites Aeqyptiaca Del Enzymatic Protein Hydrolysates

Authors: Friday A. Ogori, Ojotu M. Eke, Oneh J. Abu, Abraham T. Girgih

Abstract:

B.aeqygtiaca del (Balanites aegyptiaca del) seed protein concentrate (APC) was hydrolyzed using different enzymes such as pepsin+pancreatin (PP), Alcalase (Alca), and Flavourzyme (Flav). The Alca has higher yield (100%) when compared to PP (83.23%) and Flav (62.90%). The hydrophobic amino acid and Sulphur containing amino acid (40.19%, 7.04%) in PP hydrolysate were higher compared to Alcalase (38.92%, 6.69%), Flavourenzyme (37.43%,6.35%), and APC (39.97%, 6.95%) samples. The PP has stronger DPPH, Hydroxyl radical quenching, Ferric reducing activity, and linoleic acid peroxidation activity, followed by the protein concentrate (APC) and Alcalase (Alca), while Flavourenzyme (Flav) derived hydrolysate was least in scavenging and inhibiting radical peroxidation properties. Flavourenzyme derived hydrolysate had stronger Ferric reducing antioxidant potential and metal chelating property. The result showed that the Alcalase hydrolysate has promising peptide yield, and PP hydrolysate had excellent amino acid residues and good in-vitro antioxidant potentials and could be a preferred ingredients in the nutraceutical and functional food emerging industries.

Keywords: balanites aegyptiaca del, protein concentrate, protein hydrolysates, enzymatic hydrolysis, antioxidants

Procedia PDF Downloads 40
137 A Recombinant Group a Streptococcus (GAS-2W) Strain Elicits Protective Immunity in Mice through Induction of an IFN-γ Dependent Humoral Response

Authors: Shiva Emami, Jenny Persson, Bengt Johansson Lindbom

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Group A streptococcus (GAS) is a prevalent human pathogen, causing a wide range of infections and diseases. One of the most well-known virulence factors in GAS is M protein, a surface protein that facilitates bacterial invasion. In this study, we used a recombinant GAS strain (GAS-2W) expressing M protein containing a hyper immunogenic peptide (2W). Mice were immunized three times with heat-killed-GAS subcutaneously at three weeks intervals. Three weeks post last immunization, mice were challenged intraperitoneally with a lethal dose of live GAS. In order to investigate the impact of IFN-ƴ and antibodies in protection against GAS infection, we used a mouse model knock-out for IFN-ƴ (IFN-ƴ KO). We observed immunization with GAS-2W strain can increase protection against GAS infection in mice compared with the original GAS strain. Higher levels of antibodies against M1 protein were measured in GAS-2W-immunized mice. There was also a significant increase in IgG2c response in mice immunized with GAS2W. By using IFN-ƴ KO mice, we showed that not a high level of total IgG, but IgG2c was correlated with protection through the i.p challenge. It also emphasizes the importance of IFN-ƴ cytokine to combat GAS by isotype switching to IgG2c (which is opsonic for phagocytosis). Our data indicate the crucial role of IFN-ƴ in the protective immune response that, together with IgG2c, can induce protection against GAS.

Keywords: Group A streptococcus, IgG2c, IFN-γ, protection

Procedia PDF Downloads 59
136 Extraction of Amorphous SiO₂ From Equisetnm Arvense Plant for Synthesis of SiO₂/Zeolitic Imidazolate Framework-8 Nanocomposite and Its Photocatalytic Activity

Authors: Babak Azari, Afshin Pourahmad, Babak Sadeghi, Masuod Mokhtari

Abstract:

In this work, Equisetnm arvense plant extract was used for preparing amorphous SiO₂. For preparing of SiO₂/zeolitic imidazolate framework-8 (ZIF-8) nanocomposite by solvothermal method, the synthesized SiO₂ was added to the synthesis mixture ZIF-8. The nanocomposite was characterized using a range of techniques. The photocatalytic activity of SiO₂/ZIF-8 was investigated systematically by degrading crystal violet as a cationic dye under Ultraviolet light irradiation. Among synthesized samples (SiO₂, ZIF-8 and SiO₂/ZIF-8), the SiO₂/ZIF-8 exhibited the highest photocatalytic activity and improved stability compared to pure SiO₂ and ZIF-8. As evidenced by Scanning Electron Microscopy and Transmission electron microscopy images, ZIF-8 particles without aggregation are located over SiO₂. The SiO₂ not only provides structured support for ZIF-8 but also prevents the aggregation of ZIF-8 Metal-organic framework in comparison to the isolated ZIF-8. The superior activity of this photocatalyst was attributed to the synergistic effects from SiO₂ owing to (I) an electron acceptor (from ZIF-8) and an electron donor (to O₂ molecules), (II) preventing recombination of electron-hole in ZIF-8, and (III) maximum interfacial contact ZIF-8 with the SiO₂ surface without aggregation or prevent the accumulation of ZIF-8. The results demonstrate that holes (h+) and •O₂- are primary reactive species involved in the photocatalytic oxidation process. Moreover, the SiO₂/ZIF-8 photocatalyst did not show any obvious loss of photocatalytic activity during five-cycle tests, which indicates that the heterostructured photocatalyst was highly stable and could be used repeatedly.

Keywords: nano, zeolit, potocatalist, nanocomposite

Procedia PDF Downloads 51
135 Application of Synthetic Monomers Grafted Xanthan Gum for Rhodamine B Removal in Aqueous Solution

Authors: T. Moremedi, L. Katata-Seru, S. Sardar, A. Bandyopadhyay, E. Makhado, M. Joseph Hato

Abstract:

The rapid industrialisation and population growth have led to a steady fall in freshwater supplies worldwide. As a result, water systems are affected by modern methods upon use due to secondary contamination. The application of novel adsorbents derived from natural polymer holds a great promise in addressing challenges in water treatment. In this study, the UV irradiation technique was used to prepare acrylamide (AAm) monomer, and acrylic acid (AA) monomer grafted xanthan gum (XG) copolymer. Furthermore, the factors affecting rhodamine B (RhB) adsorption from aqueous media, such as pH, dosage, concentration, and time were also investigated. The FTIR results confirmed the formation of graft copolymer by the strong vibrational bands at 1709 cm-1 and 1612 cm-1 for AA and AAm, respectively. Additionally, more irregular, porous and wrinkled surface observed from SEM of XG-g-AAm/AA indicated copolymerization interaction of monomers. The optimum conditions for removing RhB dye with a maximum adsorption capacity of 313 mg/g at 25 0C from aqueous solution were pH approximately 5, initial dye concentration = 200 ppm, adsorbent dose = 30 mg. Also, the detailed investigation of the isothermal and adsorption kinetics of RhB from aqueous solution showed that the adsorption of the dye followed a Freundlich model (R2 = 0.96333) and pseudo-second-order kinetics. The results further indicated that this absorbent based on XG had the universality to remove dye through the mechanism of chemical adsorption. The outstanding adsorption potential of the grafted copolymer could be used to remove cationic dyes from aqueous solution as a low-cost product.

Keywords: xanthan gum, adsorbents, rhodamine B, Freundlich

Procedia PDF Downloads 103
134 Modifiable Poly Methacrylic Acid-Co-Acrylonitrile Microgels Fabricated with Cu and Co Nanoparticles for Simultaneous Catalytic Reduction of Multiple Compounds

Authors: Muhammad Ajmal, Muhammad Siddiq, Nurettin Sahiner

Abstract:

We prepared poly(methacrylic acid-co-acrylonitrile) (p(MAc-co-AN)) microgels by inverse suspension polymerization, and converted the nitrile groups into amidoxime groups to obtain more hydrophilic amidoximated poly(methacrylic acid-co-acrylonitile) (amid-p(MAc-co-AN)) microgels. Amid-microgels were used as microreactors for in situ synthesis of copper and cobalt nanoparticles. Cu (II) and Co (II) ions were loaded into microgels from their aqueous metal salt solutions and then converted to corresponding metal nanoparticle (MNP) by treating the loaded metal ions with sodium borohydride (NaBH4). The characterization of the prepared microgels and microgel metal nanoparticle composites was carried out by SEM, TEM and TG analysis. The amounts of metal nanoparticles within microgels were estimated by AAS measurements by dissolving the MNP entrapped within microgels by concentrated HCl acid treatment. Catalytic performances of the prepared amid-p(MAc-co-AN)-M (M: Cu, Co) microgel composites were investigated by using them as catalyst for the degradation of cationic and anionic organic dyes such as eosin Y (EY), methylene blue (MB) and methyl Orange (MO), and for the reduction of nitro aromatic pollutants like 2-nitrophenol (2-NP) and 4-nitrophenol (4-NP) to their corresponding amino phenols. Here, we also report for the first time, the simultaneous degradation/reduction of MB, EY, and 4-NP by amid-p(MAc-co-AN)-Cu microgel composites. Different parameters affecting the reduction rates such as metal types, amount of catalysts, temperature and the amount of reducing agent were investigated.

Keywords: microgels, nanoparticles, catalyst, pollutants

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133 An Integrative Computational Pipeline for Detection of Tumor Epitopes in Cancer Patients

Authors: Tanushree Jaitly, Shailendra Gupta, Leila Taher, Gerold Schuler, Julio Vera

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Genomics-based personalized medicine is a promising approach to fight aggressive tumors based on patient's specific tumor mutation and expression profiles. A remarkable case is, dendritic cell-based immunotherapy, in which tumor epitopes targeting patient's specific mutations are used to design a vaccine that helps in stimulating cytotoxic T cell mediated anticancer immunity. Here we present a computational pipeline for epitope-based personalized cancer vaccines using patient-specific haplotype and cancer mutation profiles. In the workflow proposed, we analyze Whole Exome Sequencing and RNA Sequencing patient data to detect patient-specific mutations and their expression level. Epitopes including the tumor mutations are computationally predicted using patient's haplotype and filtered based on their expression level, binding affinity, and immunogenicity. We calculate binding energy for each filtered major histocompatibility complex (MHC)-peptide complex using docking studies, and use this feature to select good epitope candidates further.

Keywords: cancer immunotherapy, epitope prediction, NGS data, personalized medicine

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132 Comparison of Physicochemical Properties of DNA-Ionic Liquids Complexes

Authors: Ewelina Nowak, Anna Wisla-Swider, Gohar Khachatryan, Krzysztof Danel

Abstract:

Complexes of ionic liquids with different heterocyclic-rings were synthesized by ion exchange reactions with pure salmon DNA. Ionic liquids (ILs) like 1-hexyl-3-methylimidazolium chloride, 1-butyl-4-methylpyridinium chloride and 1-ethyl-1-methylpyrrolidinium bromide were used. The ILs were built into helical state and confirmed by IR spectrometric techniques. Patterns of UV-Vis, photoluminescence, IR, and CD spectra indicated inclusion of small molecules into DNA structure. Molecular weight and radii of gyrations values of ILs-DNA complexes chains were established by HPSEC–MALLS–RI method. Modification DNA with 1-ethyl-1-methylpyrrolidinium bromide gives more uniform material and leads to elimination of high molecular weight chains. Thus, the incorporation DNA double helical structure with both 1-hexyl-3-methylimidazolium chloride and 1-butyl-4-methylpyridinium chloride exhibited higher molecular weight values. Scanning electron microscopy images indicate formation of nanofibre structures in all DNA complexes. Fluorescence depends strongly on the environment in which the chromophores are inserted and simultaneously on the molecular interactions with the biopolymer matrix. The most intensive emission was observed for DNA-imidazole ring complex. Decrease in intensity UV-Vis peak absorption is a consequence of a reduction in the spatial order of polynucleotide strands and provides different π–π stacking structure. Changes in optical properties confirmed by spectroscopy methods make DNA-ILs complexes potential biosensor applications.

Keywords: biopolymers, biosensors, cationic surfactant, DNA, DNA-gels

Procedia PDF Downloads 161
131 Surface Active Phthalic Acid Ester Produced by a Rhizobacterial Strain

Authors: M. L. Ibrahim, A. Abdulhamid

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A surface active molecule synthesized by a rhizobacterial strain Bacillus lentus isolated from Cajanus cajan was investigated. The bioemulsifier was extracted, purified and partially characterized using standard methods. Surface properties of the bioemulsifier were determined by studying the emulsification index, solubility test and stability studies. Partial purification of the bioemulsifier was carried out using FT-IR analysis, Silica-gel column chromatography and thin layer chromatography. GC-MS analysis was carried out to detect the composition and mass of the lipids and esters. The isolate showed an emulsifying activity of 57% and surface activity of 36mm. The stability studies revealed that the bioemulsifier had better stability at temperature of 70oC, 8% pH and 8% NaCl concentration. FT-IR indicated the bioemulsifier to contain peptide and aliphatic chain, TLC revealed the compound to be ninhydrin positive and Column chromatography showed the presence of three amino acids namely; glutamine, valine and cysteine. GC-MS indicated the lipid moiety to contain aliphatic chain ranging from C9-C16 and two major peaks of 1,2-benzenedicarboxylic acid diethyl octyl ester. Therefore, surface active agent from Bacillus lentus can be used effectively in a wide range of applications such as in MEOR and in the biosynthesis of plasticizers for industrial uses.

Keywords: Bacillus lentus, bioemulsifiers, phthalic acid ester, Rhizosphere

Procedia PDF Downloads 387
130 Antimicrobial Peptide Produced by Lactococcus garvieae with a Broad Inhibition Spectrum

Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

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By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

Procedia PDF Downloads 211
129 Bile Salt Induced Microstructural Changes of Gemini Surfactant Micelles

Authors: Vijaykumar Patel, P. Bahadur

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Microstructural evolution of a cationic gemini surfactant 12-4-12 micelles in the presence of bile salts has been investigated using different techniques. A negative value of interaction parameter evaluated from surface tension measurements is a signature of strong synergistic interaction between oppositely charged surfactants. Both the bile salts compete with each other in inducing the micellar transition of 12-4-12 micelles depending on their hydrophobicity. Viscosity measurements disclose that loading of bile salts induces morphological changes in 12-4-12 micelles; sodium deoxycholate is more efficient in altering the aggregation behaviour of 12-4-12 micelles compared to sodium cholate and presents pronounced increase in viscosity and micellar growth which is suppressed at elevated temperatures. A remarkable growth of 12-4-12 micelles in the presence of sodium deoxycholate at low pH has been ascribed to the solubilization of bile acids formed in acidic medium. Small angle neutron scattering experiments provided size and shape of 12-4-12/bile salt mixed micelles are explicated on the basis of hydrophobicity of bile salts. The location of bile salts in micelle was determined from nuclear overhauser effect spectroscopy. The present study characterizes 12-4-12 gemini-bile salt mixed systems which significantly enriches our knowledge, and such a structural transition provides an opportunity to use these bioamphiphiles as delivery vehicles and in some pharmaceutical formulations.

Keywords: gemini surfactants, bile salts, SANS (small angle neutron scattering), NOESY (nuclear overhauser effect spectroscopy)

Procedia PDF Downloads 127
128 Cyclic NGR Peptide Anchored Block Co-Polymeric Nanoparticles as Dual Targeting Drug Delivery System for Solid Tumor Therapy

Authors: Madhu Gupta, G. P. Agrawa, Suresh P. Vyas

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Certain tumor cells overexpress a membrane-spanning molecule aminopeptidase N (CD13) isoform, which is the receptor for peptides containing the NGR motif. NGR-modified Docetaxel (DTX)-loaded PEG-b-PLGA polymeric nanoparticles (cNGR-DNB-NPs) were developed and evaluated for their in vitro potential in HT-1080 cell line. The cNGR-DNB-NPs containing particles were about 148 nm in diameter with spherical shape and high encapsulation efficiency. Cellular uptake was confirmed both qualitatively and quantitatively by Confocal Laser Scanning Microscopy (CLSM) and flow cytometry. Both quantitatively and qualitatively results confirmed the NGR conjugated nanoparticles revealed the higher uptake of nanoparticles by CD13-overexpressed tumor cells. Free NGR inhibited the cellular uptake of cNGR-DNB-NPs, revealing the mechanism of receptor mediated endocytosis. In vitro cytotoxicity studies demonstrated that cNGR-DNB-NPs, formulation was more cytotoxic than unconjugated one, which were consistent well with the observation of cellular uptake. Hence, the selective delivery of cNGR-DNB-NPs formulation in CD13-overexpressing tumors represents a potential approach for the design of nanocarrier-based dual targeted delivery systems for targeting the tumor cells and vasculature.

Keywords: solid Tumor, docetaxel, targeting, NGR ligand

Procedia PDF Downloads 458
127 Spectrofluorometric Studies on the Interactions of Bovine Serum Albumin with Dimeric Cationic Surfactants

Authors: Srishti Sinha, Deepti Tikariha, Kallol K. Ghosh

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Over the past few decades protein-surfactant interactions have been a subject of extensive studies as they are of great importance in wide variety of industries, biological, pharmaceutical and cosmetic systems. Protein-surfactant interactions have been explored the effect of surfactants on structure of protein in the form of solubilization and denaturing or renaturing of protein. Globular proteins are frequently used as functional ingredients in healthcare and pharmaceutical products, due to their ability to catalyze biochemical reactions, to be adsorbed on the surface of some substance and to bind other moieties and form molecular aggregates. One of the most widely used globular protein is bovine serum albumin (BSA), since it has a well-known primary structure and been associated with the binding of many different categories of molecules, such as dyes, drugs and toxic chemicals. Protein−surfactant interactions are usually dependent on the surfactant features. Most of the research has been focused on single-chain surfactants. More recently, the binding between proteins and dimeric surfactants has been discussed. In present study interactions of one dimeric surfactant Butanediyl-1,4-bis (dimethylhexadecylammonium bromide) (16-4-16, 2Br-) and the corresponding single-chain surfactant cetyl trimethylammonium bromide (CTAB) with bovine serum albumin (BSA) have been investigated by surface tension and spectrofluoremetric methods. It has been found that the bindings of all gemini surfactant to BSA were cooperatively driven by electrostatic and hydrophobic interactions. The gemini surfactant carrying more charges and hydrophobic tails, showed stronger interactions with BSA than the single-chain surfactant.

Keywords: bovine serum albumin, gemini surfactants, hydrophobic interactions, protein surfactant interaction

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126 Crystallization in the TeO2 - Ta2O5 - Bi2O3 System: From Glass to Anti-Glass to Transparent Ceramic

Authors: Hasnaa Benchorfi

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The Tellurite glasses exhibit interesting properties, notably their low melting point (700-900°C), high refractive index (≈2), high transparency in the infrared region (up to 5−6 μm), interesting linear and non-linear optical properties and high rare earth ions solubility. These properties give tellurite glasses a great interest in various optical applications. Transparent ceramics present advantages compared to glasses, such as improved mechanical, thermal and optical properties. But, the elaboration process of these ceramics requires complex sintering conditions. The full crystallization of glass into transparent ceramics is an alternative to circumvent the technical challenges related to the ceramics obtained by conventional processing. In this work, a crystallization study of a specific glass composition in the system TeO2-Ta2O5-Bi2O3 shows structural transitions from the glass to the stabilization of an unreported anti-glass phase to a transparent ceramic upon heating. An anti-glass is a material with a cationic long-range order and a disordered anion sublattice. Thus, the X-ray diffraction patterns show sharp peaks, while the Raman bands are broad and similar to those of the parent glass. The structure and microstructure of the anti-glass and corresponding ceramic were characterized by Powder X-Ray Diffraction, Electron Back Scattered Diffraction, Transmission Electron Microscopy and Raman spectroscopy. The optical properties of the Er3+-doped samples are also discussed.

Keywords: glass, congruent crystallization, anti-glass, glass-ceramic, optics

Procedia PDF Downloads 47
125 A Novel Cold Asphalt Concrete Mixture for Heavily Trafficked Binder Course

Authors: Anmar Dulaimi, Hassan Al Nageim, Felicite Ruddock, Linda Seton

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Cold bituminous asphalt mixture (CBEM) provide a sustainable, cost effective and energy efficiency alternative to traditional hot mixtures. However, these mixtures have a comparatively low initial strength and as it is considered as evolutionary materials, mainly in the early life where the initial cohesion is low and builds up slowly. On the other hand, asphalt concrete is, by far, the most common mixtures in use as binder course and base in road pavement in the UK having a continuous grade offer a good aggregate interlock results in this material having very good load-spreading properties as well as a high resistance to permanent deformation. This study aims at developing a novel fast curing cold asphalt concrete binder course mixtures by using Ordinary Portland Cement (OPC) as a replacement to conventional mineral filler (0%-100%) while new by-product material (LJMU-A2) was used as a supplementary cementitious material. With this purpose, cold asphalt concrete binder course mixtures with cationic emulsions were studied by means of stiffness modulus whereas water sensitivity was approved by assessing the stiffness modulus ratio before and after sample conditioning. The results indicate that a substantial enhancement in the stiffness modulus and a considerable improvement of water sensitivity resistance by adding of LJMU-A2 to the cold asphalt mixtures as a supplementary cementitious material. Moreover, the addition of LJMU-A2 to those mixtures leads to stiffness modulus after 2- day curing comparable to those obtained with Portland cement after 7-day curing.

Keywords: cold mix asphalt, binder course, cement, stiffness modulus, water sensitivity

Procedia PDF Downloads 281
124 An Antifungal Peptide from Actinobacteria (Streptomyces Sp. TKJ2): Isolation and Partial Characterization

Authors: Abdelaziz Messis, Azzeddine Bettache, Nawel Boucherba, Said Benallaoua, Mouloud Kecha

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Actinobacteria are of special biotechnological interest since they are known to produce chemically diverse compounds with a wide range of biological activity. This distinct clade of Gram-positve bacteria include some of the key antibiotic producers and are also sources of several bioactive compounds, established commercially a newly filamentous bacteria was recovered from Tikjda forest soil (Algeria) for its high antifungal activity against various pathogenic and phytopathogenic fungi. The nucleotide sequence of the 16S rRNA gene (1454 pb) of Streptomyces sp. TKJ2 exhibited close similarity (99 %) with other Streptomyces16S rRNA genes. Antifungal metabolite production of Streptomyces sp TKJ2 was evaluated using six different fermentation media. The extracellular products contained potent antifungal agents. Antifungal protein produced by Streptomyces sp. TKJ2 on PCA medium has been purified by ammonium sulfate precipitation, SPE column chromatography and high-performance liquid chromatography in a reverse-phase column. The UV chromatograms of the active fractions obtained at 214 nm by NanoLC-ESI-MS/MS have different molecular weights. The F20 Peptidic fraction obtained from culture filtrat of Streptomyces sp. TKJ2 precipitated at 30% of ammonium sulfate was selected for analysis by infusion ESI-MS which yielded a singly charged ion mass of 437.17 Da.

Keywords: actinobacteria, antifungal protein, chromatography, Streptomyces

Procedia PDF Downloads 353
123 Efficient Mercury Sorbent: Activated Carbon and Metal Organic Framework Hybrid

Authors: Yongseok Hong, Kurt Louis Solis

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In the present study, a hybrid sorbent using the metal organic framework (MOF), UiO-66, and powdered activated carbon (pAC) is synthesized to remove cationic and anionic metals simultaneously. UiO-66 is an octahedron-shaped MOF with a Zr₆O₄(OH)₄ metal node and 1,4-benzene dicarboxylic acid (BDC) organic linker. Zr-based MOFs are attractive for trace element remediation in wastewaters, because Zr is relatively non-toxic as compared to other classes of MOF and, therefore, it will not cause secondary pollution. Most remediation studies with UiO-66 target anions such as fluoride, but trace element oxyanions such as arsenic, selenium, and antimony have also been investigated. There have also been studies involving mercury removal by UiO-66 derivatives, however these require post-synthetic modifications or have lower effective surface areas. Activated carbon is known for being a readily available, well-studied, effective adsorbent for metal contaminants. Solvothermal method was employed to prepare hybrid sorbent from UiO66 and activated carbon, which could be used to remove mercury and selenium simultaneously. The hybrid sorbent was characterized using FSEM-EDS, FT-IR, XRD, and TGA. The results showed that UiO66 and activated carbon are successfully composited. From BET studies, the hybrid sorbent has a SBET of 1051 m² g⁻¹. Adsorption studies were performed, where the hybrid showed maximum adsorption of 204.63 mg g⁻¹ and 168 mg g⁻¹ for Hg (II) and selenite, respectively, and follows the Langmuir model for both species. Kinetics studies have revealed that the Hg uptake of the hybrid is pseudo-2nd order and has rate constant of 5.6E-05 g mg⁻¹ min⁻¹ and the selenite uptake follows the simplified Elovich model with α = 2.99 mg g⁻¹ min⁻¹, β = 0.032 g mg⁻¹.

Keywords: adsorption, flue gas wastewater, mercury, selenite, metal organic framework

Procedia PDF Downloads 149
122 Cholesterol-Lowering Effects of Lactobacillus plantarum Isolated from Northeastern Thai Fermented Vegetable Brassica juncea (L.)

Authors: T. Warinpramote, J. Sanguanjeen, P. Pholphakwaen, S. Kittisorayut, J. Kerdtubtim, S. Palachote, M. Taweechotipatr

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Cholesterol is a type of lipid molecule which is the significant risk factor for coronary heart disease. Currently, there are many cholesterol-lowering alternative treatments especially bile salt hydrolase positive lactobacilli. BSH can cleave the peptide linkage of bile salt, which results in removal of the amino acid group from the steroid core and increases production of the new bile acid by using more cholesterol. The purpose of this study was to isolate, and screen probiotic characteristics of lactobacilli from fermented Thai foods and further investigated for their comparative BSH activity. The result showed that 2 of 81 Lactobacillus strains, JPK2-2 and JPK3-2, isolated from Brassica juncea (L.) had significantly exhibited high BSH activity. In addition, these Lactobacillus strains showed their results that the ability to tolerate acid and bile salt. Furthermore, the using of 16S rDNA sequencing for definitive microbial identifications showed that these 2 strains belong to Lactobacillus plantarum. In the future, the L. plantarum with BSH activity strains JPK2-2 and JPK3-2 may be the candidate probiotics for application in functional foods and dairy products to improve in the patient with cardiovascular diseases.

Keywords: Lactobacillus plantarum, probiotics, bile salt hydrolase, cholesterol-lowering, fermented Thai food

Procedia PDF Downloads 122
121 A Comparison of Generation Dependent Brain Targeting Potential of(Poly Propylene Mine) Dendrimers

Authors: Nitin Dwivedi, Jigna Shah

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Aim and objective of study: This article indicates a comparison among various generations of dendrimers, a dendrimer is a bioactive material has repetitively branched molecule and used for delivery of various therapeutic active agents. This debut report compares the effect various generations of PPI dendrimers for brain targeting and management of neurodegenerative disorders potential on single platform. This report involves the study of the various mechanism of synthesis ligand anchored various generations PPI dendrimers deliver the drug directly to the CNS, prove their effectiveness in the management of the various neurodegenerative disease. Material and Methods: The Memantine an anti-Alzheimer drug loaded in different generations (3.0G, 4.0G, and 5.0G) of PPI dendrimers which were synthesized were synthesized. The various studies investigate the effect of PPI dendrimers generation on different characteristic parameters i.e. synthesis procedure, drug loading, release behavior, hemolysis profile at different concentration, MRI study for determine the route drug from olfactory transfer, animal model study in vitro, as well as in vivo performance. The outcomes of the investigation indicate drug delivery benefit as well as superior biocompatibility of 4.0G PPI dendrimer over 3.0G and 5.0G dendrimer, respectively. Results and Conclusion: The above study indicate the superiority of in drug delivery system with maximum drug utilization and minimize the drug dose for neurodegenerative disorder over 5.0G PPI dendrimers. So, 4.0G PPI dendrimers are the safe formulations for the symptomatic treatment of the neurodegenerative disorder. The fifth-generation poly(propyleneimine) (PPI) dendrimers, inherent toxicity due to the presence of many peripheral cationic groups is the major issue that limits their applicability.

Keywords: Alzheimer disease, generation, memantine, PPI

Procedia PDF Downloads 642
120 Sitagliptin-AntiCD4 Mab Conjugated T Cell Targeting Therapy for the Effective Treatment of Type I Diabetes

Authors: T. Mahesh, M. K. Samanta

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Antibody dug conjugate (ADC’s) concept is a less explored and more trustable for the treatment of Type 1 diabetes (T1D). T1D is thought to arise from selective immunologically mediated destruction of the insulin- producing β-cells in the pancreatic islets of Langerhans with consequent insulin deficiency. It is evident that type 1 diabetes can be conquered, by 1) to stop immune destruction of βcells, 2) to replace or regenerate β-cells, and 3) to preserve β-cell function and mass. Many studies found that the regulatory T cells (Tregs) are crucial for the maintenance of immunological tolerance. Immune tolerance is liable for the activation of the Th1 response. The important role of Th1 response in pathology of T1D entails the depletion of CD4+ T cells, which initiated the use of anti-CD4 monoclonal antibodies (mAbs) against CD4+ T cells to interfere with induction of T1D.Insulin is regulated by Glucagon-Like Peptide-1 hormone (GLP-1) which also stimulates β-cells proliferation as the half-life of GLP-1 harmone is less due to rapid degradation by DPP-IV enzyme an alternative DPP-IV-inhibitors can increase the half-life of GLP-1 through which it conquers the replacement and reserve β-cells mass. Thus in the present study Anti-CD4 mAb was conjugated with Sitagliptin which is a DPP-IV inhibitor Drug loaded in Nanoparticles through Sulfo-MBS cross-linkers. The above study can be an effective approach for treatment to overcome the Passive subcutaneous insulin therapy.

Keywords: antibody drug conjugates, anti-CD4 Mab, DPP IV inhibitors, GLP-1

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119 Effects of Auxetic Antibacterial Zwitterion Carboxylate and Sulfate Copolymer Hydrogels for Diabetic Wound Healing Application

Authors: Udayakumar Vee, Franck Quero

Abstract:

Zwitterionic polymers generally have been viewed as a new class of antimicrobial and non-fouling materials. They offer a broad versatility for chemical modification and hence great freedom for accurate molecular design, which bear an equimolar number of homogenously distributed anionic and cationic groups along their polymer chains. This study explores the effectiveness of the auxetic zwitterion carboxylate/sulfonate hydrogel in the diabetic-induced mouse model. A series of silver metal-doped auxetic zwitterion carboxylate/sulfonate/vinylaniline copolymer hydrogels is designed via a 3D printer. Zwitterion monomers have been characterized by FT-IR and NMR techniques. The effect of changing the monomers and different loading ratios of Ag over zwitterion on the final hydrogel materials' antimicrobial properties and biocompatibility will be investigated in detail. The synthesized auxetic hydrogel has been characterized using a wide range of techniques to help establish the relationship between molecular level and macroscopic properties of these materials, including mechanical and antibacterial and biocompatibility and wound healing ability. This work's comparative studies and results provide new insights and guide us in choosing a better auxetic structured material for a broad spectrum of wound healing applications in the animal model. We expect this approach to provide a versatile and robust platform for biomaterial design that could lead to promising treatments for wound healing applications.

Keywords: auxetic, zwitterion, carboxylate, sulfonate, polymer, wound healing

Procedia PDF Downloads 109
118 Antibacterial Zwitterion Carboxylate and Sulfonate Copolymer Auxetic Hydrogels for Diabetic Wound Healing Application

Authors: Udayakumar Veerabagu, Franck Quero

Abstract:

Zwitterion carboxylate and sulfonate polymers generally have been viewed as a new class of antimicrobial and non-fouling materials. They offer a broad versatility for chemical modification and hence great freedom for accurate molecular design, which bear an equimolar number of homogenously distributed anionic and cationic groups along their polymer chains. This study explores the effectiveness of the auxetic zwitterion carboxylate/sulfonate hydrogel in the diabetic-induced mouse model. A series of silver metal-doped auxetic zwitterion carboxylate/sulfonate/vinylaniline copolymer hydrogels is designed via a 3D printer. Zwitterion monomers have been characterized by FT-IR and NMR techniques. The effect of changing the monomers and different loading ratios of Ag over zwitterion on the final hydrogel materials' antimicrobial properties and biocompatibility will be investigated in detail. The synthesized auxetic hydrogel has been characterized using a wide range of techniques to help establish the relationship between molecular level and macroscopic properties of these materials, including mechanical and antibacterial and biocompatibility and wound healing ability. This work's comparative studies and results provide new insights and guide us in choosing a better auxetic structured material for a broad spectrum of wound healing applications in the animal model. We expect this approach to provide a versatile and robust platform for biomaterial design that could lead to promising treatments for wound healing applications.

Keywords: auxetic, zwitterion, carboxylate, sulfonate, polymer, wound healing

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117 Biological Activities of Flaxseed Peptides (Linusorbs)

Authors: Youn Young Shim, Ji Hye Kim, Jae Youl Cho, Martin J. T. Reaney

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Flaxseed (Linum usitatissimum L.) is gaining popularity in the food industry as a superfood due to its health-promoting properties. The flax plant synthesizes an array of biologically active cyclic peptides or linusorbs (LOs, a.k.a. cyclolinopeptides) from three or more ribosome-derived precursors. [1–9-NαC]-linusorb B3 and [1–9-NαC]-linusorb B2, suppress immunity, induce apoptosis in human epithelial cancer cell line (Calu-3) cells, and inhibit T-cell proliferation, but the mechanism of LOs action is unknown. Using gene expression analysis in nematode cultures and human cancer cell lines, we have observed that LOs exert their activity, in part, through induction of apoptosis. Specific LOs’ properties include: 1) distribution throughout the body after flaxseed consumption; 2) induce heat shock protein (HSP) 70A production as an indicator of stress and address the issue in Caenorhabditis elegans (exposure of nematode cultures to [1–9-NαC]-linusorb B3 induced a 30% increase in production of the HSP 70A protein); 3) induce apoptosis in Calu-3 cells; and 4) modulate regulatory genes in microarray analysis. These diverse activities indicate that LOs might induce apoptosis in cancer cells or act as versatile platforms to deliver a variety of biologically active molecules for cancer therapy.

Keywords: flaxseed, linusorb, cyclic peptide, orbitides, heat shock protein, apoptosis, anti-cancer

Procedia PDF Downloads 109
116 Bulk Amounts of Linear and Cyclic Polypeptides on Our Hand within a Short Time

Authors: Yu Zhang, Il Kim

Abstract:

Polypeptides with defined peptide sequences illustrate the power of remarkable applications in drug delivery, tissue engineering, sensing and catalysis. Especially the cyclic polypeptides, the distinctive topological architecture imparts many characteristic properties comparing to linear polypeptides. Here, a facile and highly efficient strategy for the synthesis of linear and cyclic polypeptides is reported using N-heterocyclic carbenes (NHCs)-mediated ring-opening polymerization (ROP) of α-amino acid N-carboxyanhydrides (NCA) in the presence or absence of primary amine initiator. The polymerization proceeds rapidly in a quasi-living manner, allowing access to linear and cyclic polypeptides of well-defined chain length and narrow polydispersity, as evidenced by nuclear magnetic resonance spectrum (1H NMR and 13C NMR spectra) and size exclusion chromatography (SEC) analysis. The cyclic architecture of the polypeptides was further verified by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectra (MALDI-TOF MS) and electrospray ionization (ESI) mass spectra, as well as viscosity studies. This approach can also simplify workup procedures and make bulk scale synthesis possible, which thereby opens avenues for practical uses in diverse areas, opening up the new generation of polypeptide synthesis.

Keywords: α-amino acid N-carboxyanhydrides, living polymerization, polypeptides, N-heterocyclic carbenes, ring-opening polymerization

Procedia PDF Downloads 136
115 Fluorescent Ph-Sensing Bandage for Point-of-Care Wound Diagnostics

Authors: Cherifi Katia, Al-Hawat Marie-Lynn, Tricou Leo-Paul, Lamontagne Stephanie, Tran Minh, Ngu Amy Ching Yie, Manrique Gabriela, Guirguis Natalie, Machuca Parra Arturo Israel, Matoori Simon

Abstract:

Diabetic foot ulcers (DFUs) are a serious and prevalent complication of diabetes. Current diagnostic options are limited to macroscopic wound analysis such as wound size, depth, and infection. Molecular diagnostics promise to improve DFU diagnosis, staging, and assessment of treatment response. Here, we developed a rapid and easy-to-use fluorescent pH-sensing bandage for wound diagnostics. In a fluorescent dye screen, we identified pyranine as the lead compound due to its suitable pH-sensing properties in the clinically relevant pH range of 6 to 9. To minimize the release of this dye into the wound bed, we screened a library of ionic microparticles and found a strong adhesion of the anionic dye to a cationic polymeric microparticle. These dye-loaded microparticles showed a strong fluorescence response in the clinically relevant pH range of 6 to 9 and a dye release below 1% after one day in biological media. The dye-loaded microparticles were subsequently encapsulated in a calcium alginate hydrogel to minimize the interaction of the microparticles with the wound tissue. This pH-sensing diagnostic wound dressing was tested on full-thickness dorsal wounds of mice, and a linear fluorescence response (R2 = 0.9909) to clinically relevant pH values was observed. These findings encourage further development of this pH-sensing system for molecular diagnostics in DFUs.

Keywords: wound ph, fluorescence, diagnostics, diabetic foot ulcer, wound healing, chronic wounds, diabetes

Procedia PDF Downloads 49
114 Bioinformatic Study of Follicle Stimulating Hormone Receptor (FSHR) Gene in Different Buffalo Breeds

Authors: Hamid Mustafa, Adeela Ajmal, Kim EuiSoo, Noor-ul-Ain

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World wild, buffalo production is considered as most important component of food industry. Efficient buffalo production is related with reproductive performance of this species. Lack of knowledge of reproductive efficiency and its related genes in buffalo species is a major constraint for sustainable buffalo production. In this study, we performed some bioinformatics analysis on Follicle Stimulating Hormone Receptor (FSHR) gene and explored the possible relationship of this gene among different buffalo breeds and with other farm animals. We also found the evolution pattern for this gene among these species. We investigate CDS lengths, Stop codon variation, homology search, signal peptide, isoelectic point, tertiary structure, motifs and phylogenetic tree. The results of this study indicate 4 different motif in this gene, which are Activin-recp, GS motif, STYKc Protein kinase and transmembrane. The results also indicate that this gene has very close relationship with cattle, bison, sheep and goat. Multiple alignment (MA) showed high conservation of motif which indicates constancy of this gene during evolution. The results of this study can be used and applied for better understanding of this gene for better characterization of Follicle Stimulating Hormone Receptor (FSHR) gene structure in different farm animals, which would be helpful for efficient breeding plans for animal’s production.

Keywords: buffalo, FSHR gene, bioinformatics, production

Procedia PDF Downloads 504
113 Development of Chitosan/Dextran Gelatin Methacrylate Core/Shell 3D Scaffolds and Protein/Polycaprolactone Melt Electrowriting Meshes for Tissue Regeneration Applications

Authors: J. D. Cabral, E. Murray, P. Turner, E. Hewitt, A. Ali, M. McConnell

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Worldwide demand for organ replacement and tissue regeneration is progressively increasing. Three-dimensional (3D) bioprinting, where a physical construct is produced using computer-aided design, is a promising tool to advance the tissue engineering and regenerative medicine fields. In this paper we describe two different approaches to developing 3D bioprinted constructs for use in tissue regeneration. Bioink development is critical in achieving the 3D biofabrication of functional, regenerative tissues. Hydrogels, cross-linked macromolecules that absorb large amounts of water, have received widespread interest as bioinks due to their relevant soft tissue mechanics, biocompatibility, and tunability. In turn, not only is bioink optimisation crucial, but the creation of vascularized tissues remains a key challenge for the successful fabrication of thicker, more clinically relevant bioengineered tissues. Among the various methodologies, cell-laden hydrogels are regarded as a favorable approach; and when combined with novel core/shell 3D bioprinting technology, an innovative strategy towards creating new vessel-like structures. In this work, we investigate this cell-based approach by using human umbilical endothelial cells (HUVECs) entrapped in a viscoelastic chitosan/dextran (CD)-based core hydrogel, printed simulataneously along with a gelatin methacrylate (GelMA) shell. We have expanded beyond our previously reported FDA approved, commercialised, post-surgical CD hydrogel, Chitogel®, by functionalizing it with cell adhesion and proteolytic peptides in order to promote bone marrow-derived mesenchymal stem cell (immortalized BMSC cell line, hTERT) and HUVECs growth. The biocompatibility and biodegradability of these cell lines in a 3D bioprinted construct is demonstrated. Our studies show that particular peptide combinations crosslinked within the CD hydrogel was found to increase in vitro growth of BMSCs and HUVECs by more than two-fold. These gels were then used as a core bioink combined with the more mechanically robust, UV irradiated GelMA shell bioink, to create 3D regenerative, vessel-like scaffolds with high print fidelity. As well, microporous MEW scaffolds made from milk proteins blended with PCL were found to show promising bioactivity, exhibiting a significant increase in keratinocyte (HaCaTs) and fibroblast (normal human dermal fibroblasts, NhDFs) cell migration and proliferation when compared to PCL only scaffolds. In conclusion, our studies indicate that a peptide functionalized CD hydrogel bioink reinforced with a GelMA shell is biocompatible, biodegradable, and an appropriate cell delivery vehicle in the creation of regenerative 3D constructs. In addition, a novel 3D printing technique, melt electrowriting (MEW), which allows fabrication of micrometer fibre meshes, was used to 3D print polycaprolactone (PCL) and bioactive milk protein, lactorferrin (LF) and whey protein (WP), blended scaffolds for potential skin regeneration applications. MEW milk protein/PCL scaffolds exhibited high porosity characteristics, low overall biodegradation, and rapid protein release. Human fibroblasts and keratinocyte cells were seeded on to the scaffolds. Scaffolds containing high concentrations of LF and combined proteins (LF+WP) showed improved cell viability over time as compared to PCL only scaffolds. This research highlights two scaffolds made using two different 3D printing techniques using a combination of both natural and synthetic biomaterial components in order to create regenerative constructs as potential chronic wound treatments.

Keywords: biomaterials, hydrogels, regenerative medicine, 3D bioprinting

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112 Comparative Studies on Spontaneous Imbibition of Surfactant/Alkaline Solution in Carbonate Rocks

Authors: M. Asgari, N. Heydari, N. Shojai Kaveh, S. N. Ashrafizadeh

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Chemical flooding methods are having importance in enhanced oil recovery to recover the trapped oil after conventional recovery, as conventional oil resources become scarce. The surfactant/alkaline process consists of injecting alkali and synthetic surfactant. The addition of surfactant to injected water reduces oil/water IFT and/or alters wettability. The alkali generates soap in situ by reaction between the alkali and naphthenic acids in the crude oil. Oil recovery in fractured reservoirs mostly depends on spontaneous imbibition (SI) of brine into matrix blocks. Thus far, few efforts have been made toward understanding the relative influence of capillary and gravity forces on the fluid flow. This paper studies the controlling mechanisms of spontaneous imbibition process in chalk formations by consideration of type and concentration of surfactants, CMC, pH and alkaline reagent concentration. Wetting properties of carbonate rock have been investigated by means of contact-angle measurements. Interfacial-tension measurements were conducted using spinning drop method. Ten imbibition experiments were conducted in atmospheric pressure and various temperatures from 30°C to 50°C. All experiments were conducted above the CMC of each surfactant. The experimental results were evaluated in terms of ultimate oil recovery and reveal that wettability alteration achieved by nonionic surfactant, which led to imbibition of brine sample containing the nonionic surfactant, while IFT value was not in range of ultra low. The displacement of oil was initially dominated by capillary forces. However, for cationic surfactant, gravity forces was the dominant force for oil production by surfactant solution to overcome the negative capillary pressure.

Keywords: alkaline, capillary, gravity, imbibition, surfactant, wettability

Procedia PDF Downloads 186
111 Synthesis of Montmorillonite/CuxCd1-xS Nanocomposites and Their Application to the Photodegradation of Methylene Blue

Authors: H. Boukhatem, L. Djouadi, H. Khalaf, R. M. Navarro, F. V. Ganzalez

Abstract:

Synthetic organic dyes are used in various industries, such as textile industry, leather tanning industry, paper production, hair dye production, etc. Wastewaters containing these dyes may be harmful to the environment and living organisms. Therefore, it is very important to remove or degrade these dyes before discharging them into the environment. In addition to standard technologies for the degradation and/or removal of dyes, several new specific technologies, the so-called advanced oxidation processes (AOPs), have been developed to eliminate dangerous compounds from polluted waters. AOPs are all characterized by the same chemical feature: production of radicals (•OH) through a multistep process, although different reaction systems are used. These radicals show little selectivity of attack and are able to oxidize various organic pollutants due to their high oxidative capacity (reduction potential of HO• Eo = 2.8 V). Heterogeneous photocatalysis, as one of the AOPs, could be effective in the oxidation/degradation of organic dyes. A major advantage of using heterogeneous photocatalysis for this purpose is the total mineralization of organic dyes, which results in CO2, H2O and corresponding mineral acids. In this study, nanomaterials based on montmorillonite and CuxCd1-xS with different Cu concentration (0.3 < x < 0.7) were utilized for the degradation of the commercial cationic textile dye Methylene blue (MB), used as a model pollutant. The synthesized nanomaterials were characterized by fourier transform infrared (FTIR) and thermogravimetric-differential thermal analysis (TG–DTA). Test results of photocatalysis of methylene blue under UV-Visible irradiation show that the photoactivity of nanomaterials montmorillonite/ CuxCd1-xS increases with the increasing of Cu concentration. The kinetics of the degradation of the MB dye was described with the Langmuir–Hinshelwood (L–H) kinetic model.

Keywords: heterogeneous photocatalysis, methylene blue, montmorillonite, nanomaterial

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110 Visualizing Matrix Metalloproteinase-2 Activity Using Extracellular Matrix-Immobilized Fluorescence Resonance Energy Transfer Bioprobe in Cancer Cells

Authors: Hawon Lee, Young-Pil Kim

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Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.

Keywords: collagen, ECM, FRET, MMP

Procedia PDF Downloads 168