Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 902

Search results for: quorum sensing

902 Utilizing the RhlR/RhlI Quorum Sensing System to Express the ß-Galactosidase Reporter Gene by Using the N-Butanoyl Homoserine Lactone and N-Hexanoyl Homoserine Lactone

Authors: Ngoc Tu Truong, Nuong T. Bui, Ben Rao, Ya L. Shen

Abstract:

Quorum sensing is a phenomenon present in many gram-negative bacteria that allows bacterial communication and controlled expression of a large suite of genes through quorum sensing signals - N-acyl homoserine lactones (AHLs). In order to investigate the ability of the rhlR/rhlI quorum sensing system in Pseudomonas aeruginosa to express the ß-Galactosidase reporter gene, an engineered E. coli strain EpHL02, was genetically engineered. This engineered E. coli strain EpHL02 responded to the presence of the N-butanoyl homoserine lactone and N-hexanoyl homoserine lactone to express the ß-Galactosidase reporter gene at a concentration limit of 5x10⁻⁸ M. This was also found to be comparable to AHLs extraction from Serratia marcescens H31. Moreover, we examined this ability of this engineered E. coli strain for respond of AHLs from extractions of Pseudomonas aeruginosa ATCC9027. The results demonstrated that the rhlR/rhlI quorum sensing system can express the ß-Galactosidase reporter gene by using the N-butanoyl homoserine lactone, N-hexanoyl homoserine lactone and AHLs from extractions of Serratia marcescens H31 and Pseudomonas aeruginosa ATCC9027 in the engineered E. coli strain EpHL02.

Keywords: N-butanoyl homoserine lactone, C4-HSL, N-hexanoyl homoserine lactone, C6-HSL, Pseudomonas aeruginosa, quorum sensing, Serratia marcescens, ß-galactosidase reporter gene

Procedia PDF Downloads 233
901 Quorum Quenching Activities of Bacteria Isolated from Red Sea Sediments

Authors: Zahid Rehman, TorOve Leiknes

Abstract:

Quorum sensing (QS) is the process by which bacteria communicate with each other through small signaling molecules, such as N-acylhomoserine lactones (AHLs). Also, certain bacteria have the ability to degrade AHL molecules by a process referred to as quorum quenching (QQ); therefore, QQ can be used to control bacterial infections and biofilm formation. In this study, we aimed to identify new species of bacteria with QQ activities. To achieve this, sediments from Red Sea were collected either in the close vicinity of Sea grass or from area with no vegetation. From these samples, we isolated 72 bacterial strains and tested their ability to degrade/inactivate AHL molecules. Chromobacterium violaceum based bioassay was used in initial screening of isolates for QQ activity. The QQ activity of the positive isolates was further confirmed and quantified by employing liquid chromatography and mass spectrometry. These analyses showed that isolated bacterial strain could degrade AHL molecules with different acyl chain length and modifications. Sequencing of 16S-rRNA genes of positive isolates revealed that they belong to three different genera. Specifically, two isolates belong to genus Erythrobacter, four to Labrenzia and one isolate belongs to Bacterioplanes. Time course experiment showed that isolate belonging to genus Erythrobacter could degrade AHLs faster than other isolates. Furthermore, these isolates were tested for their ability to inhibit formation of biofilm and degradation of 3OXO-C12 AHLs produced by P. aeruginosa PAO1. Our results showed that isolate VG12 is better at controlling biofilm formation. This aligns with the ability of VG12 to cause at least 10-fold reduction in the amount of different AHLs tested.

Keywords: quorum sensing, biofilm, quorum quenching, anti-biofouling

Procedia PDF Downloads 92
900 Enzyme Redesign: From Metal-Dependent to Metal-Independent, a Symphony Orchestra without Concertmasters

Authors: Li Na Zhao, Arieh Warshel

Abstract:

The design of enzymes is an extremely challenging task, and this is also true for metalloenzymes. In the case of naturally evolved enzymes, one may consider the active site residues as the musicians in the enzyme orchestra, while the metal can be considered as their concertmaster. Together they catalyze reactions as if they performed a masterpiece written by nature. The Lactonase can be thought as a member of the amidohydrolase family, with two concertmasters, Fe and Zn, at its active site. It catalyzes the quorum sensing signal- N-acyl homoserine lactones (AHLs or N-AHLs)- by hydrolyzing the lactone ring. This process, known as quorum quenching, provides a strategy in the treatment of infectious diseases without introducing selection pressure. However, the activity of lactonase is metal-dependent, and this dependence hampers the clinic usage. In our study, we use the empirical valence bond (EVB) approach to evaluate the catalytic contributions decomposing them to electrostatic and other components.

Keywords: enzyme redesign, empirical valence bond, lactonase, quorum quenching

Procedia PDF Downloads 168
899 Immobilizing Quorum Sensing Inhibitors on Biomaterial Surfaces

Authors: Aditi Taunk, George Iskander, Kitty Ka Kit Ho, Mark Willcox, Naresh Kumar

Abstract:

Bacterial infections on biomaterial implants and medical devices accounts for 60-70% of all hospital acquired infections (HAIs). Treatment or removal of these infected devices results in high patient mortality and morbidity along with increased hospital expenses. In addition, with no effective strategies currently available and rapid development of antibacterial resistance has made device-related infections extremely difficult to treat. Therefore, in this project we have developed biomaterial surfaces using antibacterial compounds that inhibit biofilm formation by interfering with the bacterial communication mechanism known as quorum sensing (QS). This study focuses on covalent attachment of potent quorum sensing (QS) inhibiting compounds, halogenated furanones (FUs) and dihydropyrrol-2-ones (DHPs), onto glass surfaces. The FUs were attached by photoactivating the azide groups on the surface, and the acid functionalized DHPs were immobilized on amine surface via EDC/NHS coupling. The modified surfaces were tested in vitro against pathogenic organisms such as Staphylococcus aureus and Pseudomonas aeruginosa using confocal laser scanning microscopy (CLSM). Successful attachment of compounds on the substrates was confirmed by X-ray photoelectron spectroscopy (XPS) and contact angle measurements. The antibacterial efficacy was assessed, and significant reduction in bacterial adhesion and biofilm formation was observed on the FU and DHP coated surfaces. The activity of the coating was dependent upon the type of substituent present on the phenyl group of the DHP compound. For example, the ortho-fluorophenyl DHP (DHP-2) exhibited 79% reduction in bacterial adhesion against S. aureus and para-fluorophenyl DHP (DHP-3) exhibited 70% reduction against P. aeruginosa. The results were found to be comparable to DHP coated surfaces prepared in earlier study via Michael addition reaction. FUs and DHPs were able to retain their in vitro antibacterial efficacy after covalent attachment via azide chemistry. This approach is a promising strategy to develop efficient antibacterial biomaterials to reduce device related infections.

Keywords: antibacterial biomaterials, biomedical device-related infections, quorum sensing, surface functionalization

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898 Molecular Basis of Anti-Biofilm and Anti-Adherence Activity of Syzygium aromaticum on Streptococcus mutans: In Vitro and in Vivo Study

Authors: Mohd Adil, Rosina Khan, Asad U. Khan, Vasantha Rupasinghe HP

Abstract:

The study examined the effects of Syzygium aromaticum extracts on the virulence properties of Streptococcus mutans. The activity of glucosyltransferases in the presence of crude and diethylether fraction was reduced to 80% at concentration 78.12μg/ml and 39.06μg/ml respectively. The glycolytic pH drop by S. mutans cells was also disrupted by these extracts without affecting the bacterial viability. Microscopic analysis revealed morphological changes of the S. mutans biofilms, indicating that these plant extracts at sub-MICs could significantly affect the ability of S. mutans to form biofilm with distorted extracellular matrix. Furthermore, with the help of quantitative RT-PCR, the expression of different genes involved in adherence, quorum sensing, in the presence of these extracts were down regulated. The crude and active fractions were found effective in preventing caries development in rats. The data showed that S. aromaticum holds promise as a naturally occurring source of compounds that may prevent biofilm-related oral diseases.

Keywords: biofilm, quorum sensing, Streptococcus mutans, Syzygium aromaticum extract

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897 Effectiveness of Essential Oils as Inhibitors of Quorum Sensing Activity Using Biomonitor Strain Chromobacterium Violaceum

Authors: Ivana Cabarkapa, Zorica Tomicic, Olivera Duragic

Abstract:

Antimicrobial resistance represents one of the major challenges facing humanity in the last decades. Increasing antibiotic-resistant pathogens indicates the need for the development of alternative antibacterial drugs and new treatment strategies. One of the innovative emerging treatments in overcoming multidrug-resistant pathogens certainly represents the inhibition anti-quorum sensing system. For most of the food-borne pathogens, the expression of the virulence depends on their capability communication with other members of the population by means of quorum sensing (QS). QS represents a specific way of bacterial intercellular communication, which enabled owing to their ability to detect and to respond to cell population density by gene regulation. QS mechanisms are responsible for controls the pathogenesis, virulence luminescence, motility, sporulation and biofilm formation of many organisms by regulating gene expression. Therefore, research in this field is being an attractive target for the development of new natural antibacterial agents. Anti-QS compounds are known to have the ability to prohibit bacterial pathogenicity. Considering the importance of quorum sensing during bacterial pathogenesis, this research has been focused on evaluation anti - QS properties of four essential oils (EOs) Origanum heracleoticum, Origanum vulgare, Thymus vulgare, and Thymus serpyllum, using biomonitor strain of Chromobacterium violaceum CV026. Tests conducted on Luria Bertani agar supplemented with N hexanol DL homoserine lacton (HHL) 10µl/50ml of agar. The anti-QS potential of the EOs was assayed in a range of concentrations of 200 – 0.39 µl/ml using the disc diffusion method. EOs of Th. vulgaris and T. serpyllum were exhibited anti-QS activity indicated by a non- pigmented ring with a dilution-dependent manner. The lowest dilution of EOs T. vulgaris and T. serpyllum in which they exhibited visually detectable inhibition of violacein synthesis was 6.25 µl/ml for both tested EOs. EOs of O. heracleoticum and O. vulgare were displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by the outer non-pigmented ring, in a concentration-dependent manner. The lowest dilution of EOs of O. heracleoticum and O. vulgare in which exhibited visually detectable inhibition of violacein synthesis was 1.56 and 3.25 µl/ml, respectively. Considering that, the main constituents of the tested EOs represented by monoterpenes (carvacrol, thymol, γ-terpinene, and p-cymene), anti - QS properties of tested EOs can be mainly attributed to their activity. In particular, from the scientific literature, carvacrol and thymol show a sub-inhibitory effect against foodborne pathogens. Previous studies indicated that sub-lethal concentrations of carvacrol reduced the mobility of bacteria due to the ability of interference using QS mechanism between the bacterial cells, and thereby reducing the ability of biofilm formation The precise mechanism by which carvacrol inhibits biofilm formation is still not fully understood. Our results indicated that EOs displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by an outer non- pigmented ring with visually detectable inhibition of violacein. Preliminary results suggest that EOs represent a promising alternative for effective control of the emergence and spread of resistant pathogens.

Keywords: anti-quorum sensing activity, Chromobacterium violaceum, essential oils, violacein

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896 Molecular Implication of Interaction of Human Enteric Pathogens with Phylloplane of Tomato

Authors: Shilpi, Indu Gaur, Neha Bhadauria, Susmita Goswami, Prabir K. Paul

Abstract:

Cultivation and consumption of organically grown fruits and vegetables have increased by several folds. However, the presence of Human Enteric Pathogens on the surface of organically grown vegetables causing Gastro-intestinal diseases, are most likely due to contaminated water and fecal matter of farm animals. Human Enteric Pathogens are adapted to colonize the human gut, and also colonize plant surface. Microbes on plant surface communicate with each other to establish quorum sensing. The cross talk study is important because the enteric pathogens on phylloplane have been reported to mask the beneficial resident bacteria of plant. In the present study, HEPs and bacterial colonizers were identified using 16s rRNA sequencing. Microbial colonization patterns after interaction between Human Enteric Pathogens and natural bacterial residents on tomato phylloplane was studied. Tomato plants raised under aseptic conditions were inoculated with a mixture of Serratia fonticola and Klebsiella pneumoniae. The molecules involved in cross-talk between Human Enteric Pathogens and regular bacterial colonizers were isolated and identified using molecular techniques and HPLC. The colonization pattern was studied by leaf imprint method after 48 hours of incubation. The associated protein-protein interaction in the host cytoplasm was studied by use of crosslinkers. From treated leaves the crosstalk molecules and interaction proteins were separated on 1D SDS-PAGE and analyzed by MALDI-TOF-TOF analysis. The study is critical in understanding the molecular aspects of HEP’s adaption to phylloplane. The study revealed human enteric pathogens aggressively interact among themselves and resident bacteria. HEPs induced establishment of a signaling cascade through protein-protein interaction in the host cytoplasm. The study revealed that the adaptation of Human Enteric Pathogens on phylloplane of Solanum lycopersicum involves the establishment of complex molecular interaction between the microbe and the host including microbe-microbe interaction leading to an establishment of quorum sensing. The outcome will help in minimizing the HEP load on fresh farm produce, thereby curtailing incidences of food-borne diseases.

Keywords: crosslinkers, human enteric pathogens (HEPs), phylloplane, quorum sensing

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895 Effect of Non-Thermal Plasma, Chitosan and Polymyxin B on Quorum Sensing Activity and Biofilm of Pseudomonas aeruginosa

Authors: Alena Cejkova, Martina Paldrychova, Jana Michailidu, Olga Matatkova, Jan Masak

Abstract:

Increasing the resistance of pathogenic microorganisms to many antibiotics is a serious threat to the treatment of infectious diseases and cleaning medical instruments. It should be added that the resistance of microbial populations growing in biofilms is often up to 1000 times higher compared to planktonic cells. Biofilm formation in a number of microorganisms is largely influenced by the quorum sensing regulatory mechanism. Finding external factors such as natural substances or physical processes that can interfere effectively with quorum sensing signal molecules should reduce the ability of the cell population to form biofilm and increase the effectiveness of antibiotics. The present work is devoted to the effect of chitosan as a representative of natural substances with anti-biofilm activity and non- thermal plasma (NTP) alone or in combination with polymyxin B on biofilm formation of Pseudomonas aeruginosa. Particular attention was paid to the influence of these agents on the level of quorum sensing signal molecules (acyl-homoserine lactones) during planktonic and biofilm cultivations. Opportunistic pathogenic strains of Pseudomonas aeruginosa (DBM 3081, DBM 3777, ATCC 10145, ATCC 15442) were used as model microorganisms. Cultivations of planktonic and biofilm populations in 96-well microtiter plates on horizontal shaker were used for determination of antibiotic and anti-biofilm activity of chitosan and polymyxin B. Biofilm-growing cells on titanium alloy, which is used for preparation of joint replacement, were exposed to non-thermal plasma generated by cometary corona with a metallic grid for 15 and 30 minutes. Cultivation followed in fresh LB medium with or without chitosan or polymyxin B for next 24 h. Biofilms were quantified by crystal violet assay. Metabolic activity of the cells in biofilm was measured using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) colorimetric test based on the reduction of MTT into formazan by the dehydrogenase system of living cells. Activity of N-acyl homoserine lactones (AHLs) compounds involved in the regulation of biofilm formation was determined using Agrobacterium tumefaciens strain harboring a traG::lacZ/traR reporter gene responsive to AHLs. The experiments showed that both chitosan and non-thermal plasma reduce the AHLs level and thus the biofilm formation and stability. The effectiveness of both agents was somewhat strain dependent. During the eradication of P. aeruginosa DBM 3081 biofilm on titanium alloy induced by chitosan (45 mg / l) there was an 80% decrease in AHLs. Applying chitosan or NTP on the P. aeruginosa DBM 3777 biofilm did not cause a significant decrease in AHLs, however, in combination with both (chitosan 55 mg / l and NTP 30 min), resulted in a 70% decrease in AHLs. Combined application of NTP and polymyxin B allowed reduce antibiotic concentration to achieve the same level of AHLs inhibition in P. aeruginosa ATCC 15442. The results shown that non-thermal plasma and chitosan have considerable potential for the eradication of highly resistant P. aeruginosa biofilms, for example on medical instruments or joint implants.

Keywords: anti-biofilm activity, chitosan, non-thermal plasma, opportunistic pathogens

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894 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

Abstract:

Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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893 Capacity Optimization in Cooperative Cognitive Radio Networks

Authors: Mahdi Pirmoradian, Olayinka Adigun, Christos Politis

Abstract:

Cooperative spectrum sensing is a crucial challenge in cognitive radio networks. Cooperative sensing can increase the reliability of spectrum hole detection, optimize sensing time and reduce delay in cooperative networks. In this paper, an efficient central capacity optimization algorithm is proposed to minimize cooperative sensing time in a homogenous sensor network using OR decision rule subject to the detection and false alarm probabilities constraints. The evaluation results reveal significant improvement in the sensing time and normalized capacity of the cognitive sensors.

Keywords: cooperative networks, normalized capacity, sensing time

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892 Effect of Using a Mixture of Al2O3 Nanoparticles and 3-Aminopropyltriethoxysilane as the Sensing Membrane for Polysilicon Wire on pH Sensing

Authors: You-Lin Wu, Zong-Xian Wu, Jing-Jenn Lin, Shih-Hung Lin

Abstract:

In this work, a polysilicon wire (PSW) coated with a mixture of 3-aminopropyltriethoxysilane (r-APTES) and Al2O3 nanoparticles as the sensing membrane prepared with various Al2O3/r-APTES and dispersing agent/r-APTES ratios for pH sensing is studied. The r-APTES and dispersed Al2O3 nanoparticles mixture was directly transferred to PSW surface by solution phase deposition (SPD). It is found that using a mixture of Al2O3 nanoparticles and r-APTES as the sensing membrane help in improving the pH sensing of the PSW sensor and a 5 min SPD deposition time is the best. Dispersing agent is found to be necessary for better pH sensing when preparing the mixture of Al2O3 nanoparticles and r-APTES. The optimum condition for preparing the mixture is found to be Al2O3/r-APTES ratio of 2% and dispersing agent/r-APTES ratio of 0.3%.

Keywords: al2o3 nanoparticles, ph sensing, polysilicon wire sensor, r-aptes

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891 Reliability Factors Based Fuzzy Logic Scheme for Spectrum Sensing

Authors: Tallataf Rasheed, Adnan Rashdi, Ahmad Naeem Akhtar

Abstract:

The accurate spectrum sensing is a fundamental requirement of dynamic spectrum access for deployment of Cognitive Radio Network (CRN). To acheive this requirement a Reliability factors based Fuzzy Logic (RFL) Scheme for Spectrum Sensing has been proposed in this paper. Cognitive Radio User (CRU) predicts the presence or absence of Primary User (PU) using energy detector and calculates the Reliability factors which are SNR of sensing node, threshold of energy detector and decision difference of each node with other nodes in a cooperative spectrum sensing environment. Then the decision of energy detector is combined with Reliability factors of sensing node using Fuzzy Logic. These Reliability Factors used in RFL Scheme describes the reliability of decision made by a CRU to improve the local spectrum sensing. This Fuzzy combining scheme provides the accuracy of decision made by sensornode. The simulation results have shown that the proposed technique provide better PU detection probability than existing Spectrum Sensing Techniques.

Keywords: cognitive radio, spectrum sensing, energy detector, reliability factors, fuzzy logic

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890 Engineered Control of Bacterial Cell-to-Cell Signaling Using Cyclodextrin

Authors: Yuriko Takayama, Norihiro Kato

Abstract:

Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

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889 Performance of Nakagami Fading Channel over Energy Detection Based Spectrum Sensing

Authors: M. Ranjeeth, S. Anuradha

Abstract:

Spectrum sensing is the main feature of cognitive radio technology. Spectrum sensing gives an idea of detecting the presence of the primary users in a licensed spectrum. In this paper we compare the theoretical results of detection probability of different fading environments like Rayleigh, Rician, Nakagami-m fading channels with the simulation results using energy detection based spectrum sensing. The numerical results are plotted as P_f Vs P_d for different SNR values, fading parameters. It is observed that Nakagami fading channel performance is better than other fading channels by using energy detection in spectrum sensing. A MATLAB simulation test bench has been implemented to know the performance of energy detection in different fading channel environment.

Keywords: spectrum sensing, energy detection, fading channels, probability of detection, probability of false alarm

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888 Artificial Cells Capable of Communication by Using Polymer Hydrogel

Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

Abstract:

The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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887 Clustered Regularly Interspaced Short Palindromic Repeats Interference (CRISPRi): An Approach to Inhibit Microbial Biofilm

Authors: Azna Zuberi

Abstract:

Biofilm is a sessile bacterial accretion in which bacteria adapts different physiological and morphological behavior from planktonic form. It is the root cause of about 80% microbial infections in human. Among them, E. coli biofilms are most prevalent in medical devices associated nosocomial infections. The objective of this study was to inhibit biofilm formation by targeting LuxS gene, involved in quorum sensing using CRISPRi. luxS is a synthase, involved in the synthesis of Autoinducer-2(AI-2), which in turn guides the initial stage of biofilm formation. To implement CRISPRi system, we have synthesized complementary sgRNA to target gene sequence and co-expressed with dCas9. Suppression of luxS was confirmed through qRT-PCR. The effect of luxS gene on biofilm inhibition was studied through crystal violet assay, XTT reduction assay and scanning electron microscopy. We conclude that CRISPRi system could be a potential strategy to inhibit bacterial biofilm through mechanism base approach.

Keywords: biofilm, CRISPRi, luxS, microbial

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886 Radio-Frequency Technologies for Sensing and Imaging

Authors: Cam Nguyen

Abstract:

Rapid, accurate, and safe sensing and imaging of physical quantities or structures finds many applications and is of significant interest to society. Sensing and imaging using radio-frequency (RF) techniques, particularly, has gone through significant development and subsequently established itself as a unique territory in the sensing world. RF sensing and imaging has played a critical role in providing us many sensing and imaging abilities beyond our human capabilities, benefiting both civilian and military applications - for example, from sensing abnormal conditions underneath some structures’ surfaces to detection and classification of concealed items, hidden activities, and buried objects. We present the developments of several sensing and imaging systems implementing RF technologies like ultra-wide band (UWB), synthetic-pulse, and interferometry. These systems are fabricated completely using RF integrated circuits. The UWB impulse system operates over multiple pulse durations from 450 to 1170 ps with 5.5-GHz RF bandwidth. It performs well through tests of various samples, demonstrating its usefulness for subsurface sensing. The synthetic-pulse system operating from 0.6 to 5.6 GHz can assess accurately subsurface structures. The synthetic-pulse system operating from 29.72-37.7 GHz demonstrates abilities for various surface and near-surface sensing such as profile mapping, liquid-level monitoring, and anti-personnel mine locating. The interferometric system operating at 35.6 GHz demonstrates its multi-functional capability for measurement of displacements and slow velocities. These RF sensors are attractive and useful for various surface and subsurface sensing applications. This paper was made possible by NPRP grant # 6-241-2-102 from the Qatar National Research Fund (a member of Qatar Foundation). The statements made herein are solely the responsibility of the authors.

Keywords: RF sensors, radars, surface sensing, subsurface sensing

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885 Highly Sensitive and Selective H2 Gas Sensor Based on Pd-Pt Decorated Nanostructured Silicon Carbide Thin Films for Extreme Environment Application

Authors: Satyendra Mourya, Jyoti Jaiswal, Gaurav Malik, Brijesh Kumar, Ramesh Chandra

Abstract:

Present work describes the fabrication and sensing characteristics of the Pd-Pt decorated nanostructured silicon carbide (SiC) thin films on anodized porous silicon (PSi) substrate by RF magnetron sputtering. The gas sensing performance of Pd-Pt/SiC/PSi sensing electrode towards H2 gas under low (10–400 ppm) detection limit and high operating temperature regime (25–600 °C) were studied in detail. The chemiresistive sensor exhibited high selectivity, good sensing response, fast response/recovery time with excellent stability towards H2 at high temperature. The selectivity measurement of the sensing electrode was done towards different oxidizing and reducing gases and proposed sensing mechanism discussed in detail. Therefore, the investigated Pd-Pt/SiC/PSi structure may be a highly sensitive and selective hydrogen gas sensing electrode for deployment in extreme environment applications.

Keywords: RF Sputtering, silicon carbide, porous silicon, hydrogen gas sensor

Procedia PDF Downloads 192
884 Multifunctional Composite Structural Elements for Sensing and Energy Harvesting

Authors: Amir H. Alavi, Kaveh Barri, Qianyun Zhang

Abstract:

This study presents a new generation of lightweight and mechanically tunable structural composites with sensing and energy harvesting functionalities. This goal is achieved by integrating metamaterial and triboelectric energy harvesting concepts. Proof-of-concept polymeric beam prototypes are fabricated using 3D printing methods based on the proposed concept. Experiments and theoretical analyses are conducted to quantitatively investigate the mechanical and electrical properties of the designed multifunctional beams. The results show that these integrated structural elements can serve as nanogenerators and distributed sensing mediums without a need to incorporating any external sensing modules and electronics. The feasibility of design self-sensing and self-powering structural elements at multiscale for next generation infrastructure systems is further discussed.

Keywords: multifunctional structures, composites, metamaterial, triboelectric nanogenerator, sensors, structural health monitoring, energy harvesting

Procedia PDF Downloads 116
883 Condition Monitoring of Railway Earthworks using Distributed Rayleigh Sensing

Authors: Andrew Hall, Paul Clarkson

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Climate change is predicted to increase the number of extreme weather events intensifying the strain on Railway Earthworks. This paper describes the use of Distributed Rayleigh Sensing to monitor low frequency activity on a vulnerable earthworks sectionprone to landslides alongside a railway line in Northern Spain. The vulnerable slope is instrumented with conventional slope stability sensors allowing an assessment to be conducted of the application of Distributed Rayleigh Sensing as an earthwork condition monitoring tool to enhance the resilience of railway networks.

Keywords: condition monitoring, railway earthworks, distributed rayleigh sensing, climate change

Procedia PDF Downloads 100
882 Optimal Sensing Technique for Estimating Stress Distribution of 2-D Steel Frame Structure Using Genetic Algorithm

Authors: Jun Su Park, Byung Kwan Oh, Jin Woo Hwang, Yousok Kim, Hyo Seon Park

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For the structural safety, the maximum stress calculated from the stress distribution of a structure is widely used. The stress distribution can be estimated by deformed shape of the structure obtained from measurement. Although the estimation of stress is strongly affected by the location and number of sensing points, most studies have conducted the stress estimation without reasonable basis on sensing plan such as the location and number of sensors. In this paper, an optimal sensing technique for estimating the stress distribution is proposed. This technique proposes the optimal location and number of sensing points for a 2-D frame structure while minimizing the error of stress distribution between analytical model and estimation by cubic smoothing splines using genetic algorithm. To verify the proposed method, the optimal sensor measurement technique is applied to simulation tests on 2-D steel frame structure. The simulation tests are performed under various loading scenarios. Through those tests, the optimal sensing plan for the structure is suggested and verified.

Keywords: genetic algorithm, optimal sensing, optimizing sensor placements, steel frame structure

Procedia PDF Downloads 432
881 PSRR Enhanced LDO Regulator Using Noise Sensing Circuit

Authors: Min-ju Kwon, Chae-won Kim, Jeong-yun Seo, Hee-guk Chae, Yong-seo Koo

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In this paper, we presented the LDO (low-dropout) regulator which enhanced the PSRR by applying the constant current source generation technique through the BGR (Band Gap Reference) to form the noise sensing circuit. The current source through the BGR has a constant current value even if the applied voltage varies. Then, the noise sensing circuit, which is composed of the current source through the BGR, operated between the error amplifier and the pass transistor gate of the LDO regulator. As a result, the LDO regulator has a PSRR of -68.2 dB at 1k Hz, -45.85 dB at 1 MHz and -45 dB at 10 MHz. the other performance of the proposed LDO was maintained at the same level of the conventional LDO regulator.

Keywords: LDO regulator, noise sensing circuit, current reference, pass transistor

Procedia PDF Downloads 168
880 Integration of GIS with Remote Sensing and GPS for Disaster Mitigation

Authors: Sikander Nawaz Khan

Abstract:

Natural disasters like flood, earthquake, cyclone, volcanic eruption and others are causing immense losses to the property and lives every year. Current status and actual loss information of natural hazards can be determined and also prediction for next probable disasters can be made using different remote sensing and mapping technologies. Global Positioning System (GPS) calculates the exact position of damage. It can also communicate with wireless sensor nodes embedded in potentially dangerous places. GPS provide precise and accurate locations and other related information like speed, track, direction and distance of target object to emergency responders. Remote Sensing facilitates to map damages without having physical contact with target area. Now with the addition of more remote sensing satellites and other advancements, early warning system is used very efficiently. Remote sensing is being used both at local and global scale. High Resolution Satellite Imagery (HRSI), airborne remote sensing and space-borne remote sensing is playing vital role in disaster management. Early on Geographic Information System (GIS) was used to collect, arrange, and map the spatial information but now it has capability to analyze spatial data. This analytical ability of GIS is the main cause of its adaption by different emergency services providers like police and ambulance service. Full potential of these so called 3S technologies cannot be used in alone. Integration of GPS and other remote sensing techniques with GIS has pointed new horizons in modeling of earth science activities. Many remote sensing cases including Asian Ocean Tsunami in 2004, Mount Mangart landslides and Pakistan-India earthquake in 2005 are described in this paper.

Keywords: disaster mitigation, GIS, GPS, remote sensing

Procedia PDF Downloads 351
879 Fe-Doped Graphene Nanoparticles for Gas Sensing Applications

Authors: Shivani A. Singh, Pravin S. More

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In the present inspection, we indicate the falsification of Fe-doped graphene nanoparticles by modified Hummers method. Structural and physiochemical properties of the resulting pallets were explored with the help of ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD) and scanning electron microscopy (SEM), Photoluminescence spectroscopy (PL) for graphene sample exhibits absorption peaks ~248nm. Pure graphene shows PL peak at 348 nm. After doping of Fe with graphene the PL peak shifted from 348 nm to 332 nm. The oxidation degree, i.e. the relative amount of oxygen functional groups was estimated from the relative intensities of the oxygen related bands (ORB) in the FTIR measurements. These analyses show that this modified material can be useful for gas sensing applications and to be used in diverse areas.

Keywords: chemical doping, graphene, gas sensing, sensing

Procedia PDF Downloads 139
878 Cooperative Spectrum Sensing Using Hybrid IWO/PSO Algorithm in Cognitive Radio Networks

Authors: Deepa Das, Susmita Das

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Cognitive Radio (CR) is an emerging technology to combat the spectrum scarcity issues. This is achieved by consistently sensing the spectrum, and detecting the under-utilized frequency bands without causing undue interference to the primary user (PU). In soft decision fusion (SDF) based cooperative spectrum sensing, various evolutionary algorithms have been discussed, which optimize the weight coefficient vector for maximizing the detection performance. In this paper, we propose the hybrid invasive weed optimization and particle swarm optimization (IWO/PSO) algorithm as a fast and global optimization method, which improves the detection probability with a lesser sensing time. Then, the efficiency of this algorithm is compared with the standard invasive weed optimization (IWO), particle swarm optimization (PSO), genetic algorithm (GA) and other conventional SDF based methods on the basis of convergence and detection probability.

Keywords: cognitive radio, spectrum sensing, soft decision fusion, GA, PSO, IWO, hybrid IWO/PSO

Procedia PDF Downloads 338
877 Uniform Porous Multilayer-Junction Thin Film for Enhanced Gas-Sensing Performance

Authors: Ping-Ping Zhang, Hui-Zhang, Xu-Hui Sun

Abstract:

Highly-uniform In2O3/CuO bilayer and multilayer porous thin films were successfully fabricated using self-assembled soft template and simple sputtering deposition technique. The sensor based on the In2O3/CuO bilayer porous thin film shows obviously improved sensing performance to ethanol at the lower working temperature, compared to single layer counterpart sensors. The response of In2O3/CuO bilayer sensors exhibits nearly 3 and 5 times higher than those of the single layer In2O3 and CuO porous film sensors over the same ethanol concentration, respectively. The sensing mechanism based on p-n hetero-junction, which contributed to the enhanced sensing performance was also experimentally confirmed by a control experiment which the SiO2 insulation layer was inserted between the In2O3 and CuO layers to break the p-n junction. In addition, the sensing performance can be further enhanced by increasing the number of In2O3/CuO junction layers. The facile process can be easily extended to the fabrication of other semiconductor oxide gas sensors for practical sensing applications.

Keywords: gas sensor, multilayer porous thin films, In2O3/CuO, p-n junction

Procedia PDF Downloads 240
876 A Comparative Study on Automatic Feature Classification Methods of Remote Sensing Images

Authors: Lee Jeong Min, Lee Mi Hee, Eo Yang Dam

Abstract:

Geospatial feature extraction is a very important issue in the remote sensing research. In the meantime, the image classification based on statistical techniques, but, in recent years, data mining and machine learning techniques for automated image processing technology is being applied to remote sensing it has focused on improved results generated possibility. In this study, artificial neural network and decision tree technique is applied to classify the high-resolution satellite images, as compared to the MLC processing result is a statistical technique and an analysis of the pros and cons between each of the techniques.

Keywords: remote sensing, artificial neural network, decision tree, maximum likelihood classification

Procedia PDF Downloads 208
875 Distributed Optical Fiber Vibration Sensing Using Phase Generated Carrier Demodulation Algorithm

Authors: Zhihua Yu, Qi Zhang, Mingyu Zhang, Haolong Dai

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Distributed fiber-optic vibration sensors are gaining extensive attention, for the advantages of high sensitivity, accurate location, light weight, large-scale monitoring, good concealment, and etc. In this paper, a novel optical fiber distributed vibration sensing system is proposed, which is based on self-interference of Rayleigh backscattering with phase generated carrier (PGC) demodulation algorithm. Pulsed lights are sent into the sensing fiber and the Rayleigh backscattering light from a certain position along the sensing fiber would interfere through an unbalanced Michelson Interferometry (MI) to generate the interference light. An improved PGC demodulation algorithm is carried out to recover the phase information of the interference signal, which carries the sensing information. Three vibration events were applied simultaneously to different positions over 2000m sensing fiber and demodulated correctly. Experiments show that the spatial resolution of is 10 m, and the noise level of the Φ-OTDR system is about 10-3 rad/√Hz, and the signal to noise ratio (SNR) is about 30.34dB. This vibration measurement scheme can be applied at surface, seabed or downhole for vibration measurements or distributed acoustic sensing (DAS).

Keywords: fiber optics sensors, Michelson interferometry, MI, phase-sensitive optical time domain reflectometry, Φ-OTDR, phase generated carrier, PGC

Procedia PDF Downloads 94
874 Linear Frequency Modulation-Frequency Shift Keying Radar with Compressive Sensing

Authors: Ho Jeong Jin, Chang Won Seo, Choon Sik Cho, Bong Yong Choi, Kwang Kyun Na, Sang Rok Lee

Abstract:

In this paper, a radar signal processing technique using the LFM-FSK (Linear Frequency Modulation-Frequency Shift Keying) is proposed for reducing the false alarm rate based on the compressive sensing. The LFM-FSK method combines FMCW (Frequency Modulation Continuous Wave) signal with FSK (Frequency Shift Keying). This shows an advantage which can suppress the ghost phenomenon without the complicated CFAR (Constant False Alarm Rate) algorithm. Moreover, the parametric sparse algorithm applying the compressive sensing that restores signals efficiently with respect to the incomplete data samples is also integrated, leading to reducing the burden of ADC in the receiver of radars. 24 GHz FMCW signal is applied and tested in the real environment with FSK modulated data for verifying the proposed algorithm along with the compressive sensing.

Keywords: compressive sensing, LFM-FSK radar, radar signal processing, sparse algorithm

Procedia PDF Downloads 342
873 Modern Spectrum Sensing Techniques for Cognitive Radio Networks: Practical Implementation and Performance Evaluation

Authors: Antoni Ivanov, Nikolay Dandanov, Nicole Christoff, Vladimir Poulkov

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Spectrum underutilization has made cognitive radio a promising technology both for current and future telecommunications. This is due to the ability to exploit the unused spectrum in the bands dedicated to other wireless communication systems, and thus, increase their occupancy. The essential function, which allows the cognitive radio device to perceive the occupancy of the spectrum, is spectrum sensing. In this paper, the performance of modern adaptations of the four most widely used spectrum sensing techniques namely, energy detection (ED), cyclostationary feature detection (CSFD), matched filter (MF) and eigenvalues-based detection (EBD) is compared. The implementation has been accomplished through the PlutoSDR hardware platform and the GNU Radio software package in very low Signal-to-Noise Ratio (SNR) conditions. The optimal detection performance of the examined methods in a realistic implementation-oriented model is found for the common relevant parameters (number of observed samples, sensing time and required probability of false alarm).

Keywords: cognitive radio, dynamic spectrum access, GNU Radio, spectrum sensing

Procedia PDF Downloads 148