Search results for: atmospheric boundary layer
85 Impact of Electric Field on the Optical Properties of Hydrophilic Quantum Dots
Authors: Valentina V. Goftman, Vladislav A. Pankratov, Alexey V. Markin, Tangi Aubert, Zeger Hens, Sarah De Saeger, Irina Yu. Goryacheva
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The most important requirements for biochemical applicability of quantum dots (QDs) are: 1) the surface cap should render intact or improved optical properties; 2) mono-dispersion and good stability in aqueous phase in a wide range of pH and ionic strength values; 3) presence of functional groups, available for bioconjugation; 4) minimal impact from the environment on the QDs’ properties and, vice versa, minimal influence of the QDs’ components on the environment; and 5) stability against chemical/biochemical/physical influence. The latter is especially important for in vitro and in vivo applications. For example, some physical intracellular delivery strategies (e.g., electroporation) imply a rapid high-voltage electric field impulse in order to temporarily generate hydrophilic pores in the cell plasma membrane, necessary for the passive transportation of QDs into the cell. In this regard, it is interesting to investigate how different capping layers, which can provide high stability and sufficient fluorescent properties of QDs in a water solution, behave under these abnormal conditions. In this contribution, hydrophobic core-shell CdSe/CdS/CdZnS/ZnS QDs (λem=600 nm), produced by means of the Successive Ion Layer Adsorption and Reaction (SILAR) technique, were transferred to a water solution using two of the most commonly used methods: (i) encapsulation in an amphiphilic brush polymer based on poly(maleic anhydride-alt-1-octadecene) (PMAO) modified with polyethylene glycol (PEG) chains and (ii) silica covering. Polymer encapsulation preserves the initial ligands on the QDs’ surface owing to the hydrophobic attraction between the hydrophobic groups of the amphiphilic molecules and the surface hydrophobic groups of the QDs. This covering process allows maintaining the initial fluorescent properties, but it leads to a considerable increase of the QDs’ size. However, covering with a silica shell, by means of the reverse microemulsion method, allows maintaining both size and fluorescent properties of the initial QDs. The obtained water solutions of polymer covered and silica-coated QDs in three different concentrations were exposed to a low-voltage electric field for a short time and the fluorescent properties were investigated. It is shown that the PMAO-PEG polymer acquires some additional charges in the presence of the electric field, which causes repulsion between the polymer and the QDs’ surface. This process destroys the homogeneity of the whole amphiphilic shell and it dramatically decreases the fluorescent properties (dropping to 10% from its initial value) because of the direct contact of the QDs with the strongly oxidative environment (water). In contrast, a silica shell possesses dielectric properties which allow retaining 90% of its initial fluorescence intensity, even after a longer electric impact. Thus, silica shells are clearly a preferable covering for bio-application of QDs, because – besides the high uniform morphology, controlled size and biocompatibility – it allows protecting QDs from oxidation, even under the influence of an electric field.Keywords: electric field, polymer coating, quantum dots, silica covering, stability
Procedia PDF Downloads 45884 Monitoring and Improving Performance of Soil Aquifer Treatment System and Infiltration Basins of North Gaza Emergency Sewage Treatment Plant as Case Study
Authors: Sadi Ali, Yaser Kishawi
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As part of Palestine, Gaza Strip (365 km2 and 1.8 million habitants) is considered a semi-arid zone relies solely on the Coastal Aquifer. The coastal aquifer is only source of water with only 5-10% suitable for human use. This barely covers the domestic and agricultural needs of Gaza Strip. Palestinian Water Authority Strategy is to find non-conventional water resource from treated wastewater to irrigate 1500 hectares and serves over 100,000 inhabitants. A new WWTP project is to replace the old-overloaded Biet Lahia WWTP. The project consists of three parts; phase A (pressure line & 9 infiltration basins - IBs), phase B (a new WWTP) and phase C (Recovery and Reuse Scheme – RRS – to capture the spreading plume). Currently, phase A is functioning since Apr 2009. Since Apr 2009, a monitoring plan is conducted to monitor the infiltration rate (I.R.) of the 9 basins. Nearly 23 million m3 of partially treated wastewater were infiltrated up to Jun 2014. It is important to maintain an acceptable rate to allow the basins to handle the coming quantities (currently 10,000 m3 are pumped an infiltrated daily). The methodology applied was to review and analysis the collected data including the I.R.s, the WW quality and the drying-wetting schedule of the basins. One of the main findings is the relation between the Total Suspended Solids (TSS) at BLWWTP and the I.R. at the basins. Since April 2009, the basins scored an average I.R. of about 2.5 m/day. Since then the records showed a decreasing pattern of the average rate until it reached the lower value of 0.42 m/day in Jun 2013. This was accompanied with an increase of TSS (mg/L) concentration at the source reaching above 200 mg/L. The reducing of TSS concentration directly improved the I.R. (by cleaning the WW source ponds at Biet Lahia WWTP site). This was reflected in an improvement in I.R. in last 6 months from 0.42 m/day to 0.66 m/day then to nearly 1.0 m/day as the average of the last 3 months of 2013. The wetting-drying scheme of the basins was observed (3 days wetting and 7 days drying) besides the rainfall rates. Despite the difficulty to apply this scheme accurately a control of flow to each basin was applied to improve the I.R. The drying-wetting system affected the I.R. of individual basins, thus affected the overall system rate which was recorded and assessed. Also the ploughing activities at the infiltration basins as well were recommended at certain times to retain a certain infiltration level. This breaks the confined clogging layer which prevents the infiltration. It is recommended to maintain proper quality of WW infiltrated to ensure an acceptable performance of IBs. The continual maintenance of settling ponds at BLWWTP, continual ploughing of basins and applying soil treatment techniques at the IBs will improve the I.R.s. When the new WWTP functions a high standard effluent quality (TSS 20mg, BOD 20 mg/l, and TN 15 mg/l) will be infiltrated, thus will enhance I.R.s of IBs due to lower organic load.Keywords: soil aquifer treatment, recovery and reuse scheme, infiltration basins, North Gaza
Procedia PDF Downloads 24783 Bio-Functionalized Silk Nanofibers for Peripheral Nerve Regeneration
Authors: Kayla Belanger, Pascale Vigneron, Guy Schlatter, Bernard Devauchelle, Christophe Egles
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A severe injury to a peripheral nerve leads to its degeneration and the loss of sensory and motor function. To this day, there still lacks a more effective alternative to the autograft which has long been considered the gold standard for nerve repair. In order to overcome the numerous drawbacks of the autograft, tissue engineered biomaterials may be effective alternatives. Silk fibroin is a favorable biomaterial due to its many advantageous properties such as its biocompatibility, its biodegradability, and its robust mechanical properties. In this study, bio-mimicking multi-channeled nerve guidance conduits made of aligned nanofibers achieved by electrospinning were functionalized with signaling biomolecules and were tested in vitro and in vivo for nerve regeneration support. Silk fibroin (SF) extracted directly from silkworm cocoons was put in solution at a concentration of 10wt%. Poly(ethylene oxide) (PEO) was added to the resulting SF solution to increase solution viscosity and the following three electrospinning solutions were made: (1) SF/PEO solution, (2) SF/PEO solution with nerve growth factor and ciliary neurotrophic factor, and (3) SF/PEO solution with nerve growth factor and neurotrophin-3. Each of these solutions was electrospun into a multi-layer architecture to obtain mechanically optimized aligned nanofibrous mats. For in vitro studies, aligned fibers were treated to induce β-sheet formation and thoroughly rinsed to eliminate presence of PEO. Each material was tested using rat embryo neuron cultures to evaluate neurite extension and the interaction with bio-functionalized or non-functionalized aligned fibers. For in vivo studies, the mats were rolled into 5mm long multi-, micro-channeled conduits then treated and thoroughly rinsed. The conduits were each subsequently implanted between a severed rat sciatic nerve. The effectiveness of nerve repair over a period of 8 months was extensively evaluated by cross-referencing electrophysiological, histological, and movement analysis results to comprehensively evaluate the progression of nerve repair. In vitro results show a more favorable interaction between growing neurons and bio-functionalized silk fibers compared to pure silk fibers. Neurites can also be seen having extended unidirectionally along the alignment of the nanofibers which confirms a guidance factor for the electrospun material. The in vivo study has produced positive results for the regeneration of the sciatic nerve over the length of the study, showing contrasts between the bio-functionalized material and the non-functionalized material along with comparisons to the experimental control. Nerve regeneration has been evaluated not only by histological analysis, but also by electrophysiological assessment and motion analysis of two separate natural movements. By studying these three components in parallel, the most comprehensive evaluation of nerve repair for the conduit designs can be made which can, therefore, more accurately depict their overall effectiveness. This work was supported by La Région Picardie and FEDER.Keywords: electrospinning, nerve guidance conduit, peripheral nerve regeneration, silk fibroin
Procedia PDF Downloads 24682 Mechanical Response Investigation of Wafer Probing Test with Vertical Cobra Probe via the Experiment and Transient Dynamic Simulation
Authors: De-Shin Liu, Po-Chun Wen, Zhen-Wei Zhuang, Hsueh-Chih Liu, Pei-Chen Huang
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Wafer probing tests play an important role in semiconductor manufacturing procedures in accordance with the yield and reliability requirement of the wafer after the backend-of-the-line process. Accordingly, the stable physical and electrical contact between the probe and the tested wafer during wafer probing is regarded as an essential issue in identifying the known good die. The probe card can be integrated with multiple probe needles, which are classified as vertical, cantilever and micro-electro-mechanical systems type probe selections. Among all potential probe types, the vertical probe has several advantages as compared with other probe types, including maintainability, high probe density and feasibility for high-speed wafer testing. In the present study, the mechanical response of the wafer probing test with the vertical cobra probe on 720 μm thick silicon (Si) substrate with a 1.4 μm thick aluminum (Al) pad is investigated by the experiment and transient dynamic simulation approach. Because the deformation mechanism of the vertical cobra probe is determined by both bending and buckling mechanisms, the stable correlation between contact forces and overdrive (OD) length must be carefully verified. Moreover, the decent OD length with corresponding contact force contributed to piercing the native oxide layer of the Al pad and preventing the probing test-induced damage on the interconnect system. Accordingly, the scratch depth of the Al pad under various OD lengths is estimated by the atomic force microscope (AFM) and simulation work. In the wafer probing test configuration, the contact phenomenon between the probe needle and the tested object introduced large deformation and twisting of mesh gridding, causing the subsequent numerical divergence issue. For this reason, the arbitrary Lagrangian-Eulerian method is utilized in the present simulation work to conquer the aforementioned issue. The analytic results revealed a slight difference when the OD is considered as 40 μm, and the simulated is almost identical to the measured scratch depths of the Al pad under higher OD lengths up to 70 μm. This phenomenon can be attributed to the unstable contact of the probe at low OD length with the scratch depth below 30% of Al pad thickness, and the contact status will be being stable when the scratch depth over 30% of pad thickness. The splash of the Al pad is observed by the AFM, and the splashed Al debris accumulates on a specific side; this phenomenon is successfully simulated in the transient dynamic simulation. Thus, the preferred testing OD lengths are found as 45 μm to 70 μm, and the corresponding scratch depths on the Al pad are represented as 31.4% and 47.1% of Al pad thickness, respectively. The investigation approach demonstrated in this study contributed to analyzing the mechanical response of wafer probing test configuration under large strain conditions and assessed the geometric designs and material selections of probe needles to meet the requirement of high resolution and high-speed wafer-level probing test for thinned wafer application.Keywords: wafer probing test, vertical probe, probe mark, mechanical response, FEA simulation
Procedia PDF Downloads 5781 Creative Mapping Landuse and Human Activities: From the Inventories of Factories to the History of the City and Citizens
Authors: R. Tamborrino, F. Rinaudo
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Digital technologies offer possibilities to effectively convert historical archives into instruments of knowledge able to provide a guide for the interpretation of historical phenomena. Digital conversion and management of those documents allow the possibility to add other sources in a unique and coherent model that permits the intersection of different data able to open new interpretations and understandings. Urban history uses, among other sources, the inventories that register human activities in a specific space (e.g. cadastres, censuses, etc.). The geographic localisation of that information inside cartographic supports allows for the comprehension and visualisation of specific relationships between different historical realities registering both the urban space and the peoples living there. These links that merge the different nature of data and documentation through a new organisation of the information can suggest a new interpretation of other related events. In all these kinds of analysis, the use of GIS platforms today represents the most appropriate answer. The design of the related databases is the key to realise the ad-hoc instrument to facilitate the analysis and the intersection of data of different origins. Moreover, GIS has become the digital platform where it is possible to add other kinds of data visualisation. This research deals with the industrial development of Turin at the beginning of the 20th century. A census of factories realized just prior to WWI provides the opportunity to test the potentialities of GIS platforms for the analysis of urban landscape modifications during the first industrial development of the town. The inventory includes data about location, activities, and people. GIS is shaped in a creative way linking different sources and digital systems aiming to create a new type of platform conceived as an interface integrating different kinds of data visualisation. The data processing allows linking this information to an urban space, and also visualising the growth of the city at that time. The sources, related to the urban landscape development in that period, are of a different nature. The emerging necessity to build, enlarge, modify and join different buildings to boost the industrial activities, according to their fast development, is recorded by different official permissions delivered by the municipality and now stored in the Historical Archive of the Municipality of Turin. Those documents, which are reports and drawings, contain numerous data on the buildings themselves, including the block where the plot is located, the district, and the people involved such as the owner, the investor, and the engineer or architect designing the industrial building. All these collected data offer the possibility to firstly re-build the process of change of the urban landscape by using GIS and 3D modelling technologies thanks to the access to the drawings (2D plans, sections and elevations) that show the previous and the planned situation. Furthermore, they access information for different queries of the linked dataset that could be useful for different research and targets such as economics, biographical, architectural, or demographical. By superimposing a layer of the present city, the past meets to the present-industrial heritage, and people meet urban history.Keywords: digital urban history, census, digitalisation, GIS, modelling, digital humanities
Procedia PDF Downloads 19180 Strontium and Selenium Doped Bioceramic Incorporated Hydrogel for Faster Apatite Growth and Bone Regeneration Applications
Authors: Nonita Sarin, K.J.Singh, Anuj Kumar, Davinder Singh
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Polymeric 3D hydrogels have pivotal role in bone tissue regeneration applications. Hydrogels behave similar to the living tissues because they have large water imbibing capacity in swollen state and adjust their shape according to the tissues during tissue formation after implantation. On the other hand, hydrogels are very soft, fragile and lack mechanical strength. Incorporation of bioceramics can improve mechanical strength. Furthermore, bioceramics synthesized by sol gel technique may enhance the apatite formation and degradation rates which can lead to the increase in faster rates for new bone and tissue regeneration. Simulated body fluid (SBF) induces the poly-condensation of silanol groups which leads to formation of silica matrix and provide active sites for the precipitation of Ca2+ and PO43- ions to form apatite layer which is similar to mineral form of bone. Therefore, authors have synthesized bioceramic incorporated Polyacrylamide-carboxymethylcellulose hydrogels by free radical polymerization and bioceramic compositions of xSrO-(36-x)CaO-45SiO2-ySeO3-(12-y)P2O5-7MgO (where x=0,4 and y=0,2 mol%) were synthesized by sol gel technique. Bioceramics incorporated in polymer matrix induces quicker apatite formation during immersion in SBF by raising the pH with the release of alkaline ions during ion exchange process and the apatite formation takes place in alkaline medium. The behavior of samples PABC-0 (without bioceramics) and PABC-20 (with 20 wt% bioceramics) were evaluated by X-Ray Diffraction and FTIR. In term of bioactivity, it was observed that PABC-20 has shown hydroxyapatite (HA) formation on 1st day of immersion whereas, PABC-0 was shown apatite formation on 7th day of immersion in SBF. The rapid rate of HA growth on 1st day of immersion in SBF signifies easy regeneration of damaged bone tissues. Degradation studies have been undertaken in Phosphate Buffer Saline and PABC-20 exhibited slower degradation rate up to 9%as compared to PABC-0 up to 18%. Slower degradation rate is suitable for new tissue regeneration and cell attachment. Also, Zeta potential studies have been employed to check the surface charge and it has been observed that samples carry negative charge when immersed in SBF. In addition, the swelling test of the samples have been performed and relative swelling ratio % observed for PABC-0 is 607% and PABC-20 is 305%. This indicates that the incorporation of bioceramics leads to the filling up of the voids in between the polymer matrix which in result reduces porosity and increase the mechanical strength by filling the voids. The porosity of PABC-0 is 84% and PABC-20 is 72%. PABC-20 sample demonstrates that bioceramics incorporation reduce the porosity and improves mechanical strength. Also, maximum in vitro cell viability up to 98% with MG63 cell line has been observed which indicate that the bioceramic incorporated hydrogel(PABC-20) provide the alkaline medium which is suitable environment for cell growth.Keywords: hydrogels, hydroxyapatite, MG63 cell line, zeta potential
Procedia PDF Downloads 14079 Integrative Omics-Portrayal Disentangles Molecular Heterogeneity and Progression Mechanisms of Cancer
Authors: Binder Hans
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Cancer is no longer seen as solely a genetic disease where genetic defects such as mutations and copy number variations affect gene regulation and eventually lead to aberrant cell functioning which can be monitored by transcriptome analysis. It has become obvious that epigenetic alterations represent a further important layer of (de-)regulation of gene activity. For example, aberrant DNA methylation is a hallmark of many cancer types, and methylation patterns were successfully used to subtype cancer heterogeneity. Hence, unraveling the interplay between different omics levels such as genome, transcriptome and epigenome is inevitable for a mechanistic understanding of molecular deregulation causing complex diseases such as cancer. This objective requires powerful downstream integrative bioinformatics methods as an essential prerequisite to discover the whole genome mutational, transcriptome and epigenome landscapes of cancer specimen and to discover cancer genesis, progression and heterogeneity. Basic challenges and tasks arise ‘beyond sequencing’ because of the big size of the data, their complexity, the need to search for hidden structures in the data, for knowledge mining to discover biological function and also systems biology conceptual models to deduce developmental interrelations between different cancer states. These tasks are tightly related to cancer biology as an (epi-)genetic disease giving rise to aberrant genomic regulation under micro-environmental control and clonal evolution which leads to heterogeneous cellular states. Machine learning algorithms such as self organizing maps (SOM) represent one interesting option to tackle these bioinformatics tasks. The SOMmethod enables recognizing complex patterns in large-scale data generated by highthroughput omics technologies. It portrays molecular phenotypes by generating individualized, easy to interpret images of the data landscape in combination with comprehensive analysis options. Our image-based, reductionist machine learning methods provide one interesting perspective how to deal with massive data in the discovery of complex diseases, gliomas, melanomas and colon cancer on molecular level. As an important new challenge, we address the combined portrayal of different omics data such as genome-wide genomic, transcriptomic and methylomic ones. The integrative-omics portrayal approach is based on the joint training of the data and it provides separate personalized data portraits for each patient and data type which can be analyzed by visual inspection as one option. The new method enables an integrative genome-wide view on the omics data types and the underlying regulatory modes. It is applied to high and low-grade gliomas and to melanomas where it disentangles transversal and longitudinal molecular heterogeneity in terms of distinct molecular subtypes and progression paths with prognostic impact.Keywords: integrative bioinformatics, machine learning, molecular mechanisms of cancer, gliomas and melanomas
Procedia PDF Downloads 14878 Deterioration Prediction of Pavement Load Bearing Capacity from FWD Data
Authors: Kotaro Sasai, Daijiro Mizutani, Kiyoyuki Kaito
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Expressways in Japan have been built in an accelerating manner since the 1960s with the aid of rapid economic growth. About 40 percent in length of expressways in Japan is now 30 years and older and has become superannuated. Time-related deterioration has therefore reached to a degree that administrators, from a standpoint of operation and maintenance, are forced to take prompt measures on a large scale aiming at repairing inner damage deep in pavements. These measures have already been performed for bridge management in Japan and are also expected to be embodied for pavement management. Thus, planning methods for the measures are increasingly demanded. Deterioration of layers around road surface such as surface course and binder course is brought about at the early stages of whole pavement deterioration process, around 10 to 30 years after construction. These layers have been repaired primarily because inner damage usually becomes significant after outer damage, and because surveys for measuring inner damage such as Falling Weight Deflectometer (FWD) survey and open-cut survey are costly and time-consuming process, which has made it difficult for administrators to focus on inner damage as much as they have been supposed to. As expressways today have serious time-related deterioration within them deriving from the long time span since they started to be used, it is obvious the idea of repairing layers deep in pavements such as base course and subgrade must be taken into consideration when planning maintenance on a large scale. This sort of maintenance requires precisely predicting degrees of deterioration as well as grasping the present situations of pavements. Methods for predicting deterioration are determined to be either mechanical or statistical. While few mechanical models have been presented, as far as the authors know of, previous studies have presented statistical methods for predicting deterioration in pavements. One describes deterioration process by estimating Markov deterioration hazard model, while another study illustrates it by estimating Proportional deterioration hazard model. Both of the studies analyze deflection data obtained from FWD surveys and present statistical methods for predicting deterioration process of layers around road surface. However, layers of base course and subgrade remain unanalyzed. In this study, data collected from FWD surveys are analyzed to predict deterioration process of layers deep in pavements in addition to surface layers by a means of estimating a deterioration hazard model using continuous indexes. This model can prevent the loss of information of data when setting rating categories in Markov deterioration hazard model when evaluating degrees of deterioration in roadbeds and subgrades. As a result of portraying continuous indexes, the model can predict deterioration in each layer of pavements and evaluate it quantitatively. Additionally, as the model can also depict probability distribution of the indexes at an arbitrary point and establish a risk control level arbitrarily, it is expected that this study will provide knowledge like life cycle cost and informative content during decision making process referring to where to do maintenance on as well as when.Keywords: deterioration hazard model, falling weight deflectometer, inner damage, load bearing capacity, pavement
Procedia PDF Downloads 39077 Socio-Psychological Significance of Vandalism in the Urban Environment: Destruction, Modernization, Communication
Authors: Olga Kruzhkova, Irina Vorobyeva, Roman Porozov
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Vandalism is a common phenomenon, but its definition is still not clearly defined. In the public sense, vandalism is the blatant cases of pogroms in cemeteries, destruction of public places (regardless of whether these actions are authorized), damage to significant objects of culture and history (monuments, religious buildings). From a legal point of view, only such an act can be called vandalism, which is aimed at 'desecrating buildings or other structures, damaging property on public transport or in other public places'. The key here is the notion of public property that is being damaged. In addition, the principal is the semantics of messages, expressed in a kind of sign system (drawing, inscription, symbol), which initially threatens public order, the calmness of citizens, public morality. Because of this, the legal qualification of vandalism doesn’t include a sufficiently wide layer of environmental destructions that are common in modern urban space (graffiti and other damage to private property, broken shop windows, damage to entrances and elevator cabins), which in ordinary consciousness are seen as obvious facts of vandalism. At the same time, the understanding of vandalism from the position of psychology implies an appeal to the question of the limits of the activity of the subject of vandalism and his motivational basis. Also recently, the discourse on the positive meaning of some forms of vandalism (graffiti, street-art, etc.) has been activated. But there is no discussion of the role and significance of vandalism in public and individual life, although, like any socio-cultural and socio-psychological phenomenon, vandalism is not groundless and meaningless. Our aim of the study was to identify and describe the functions of vandalism as a socio-cultural and socio-psychological phenomenon of the life of the urban community, as well as personal determinants of its manifestations. The study was conducted in the spatial environment of the Russian megalopolis (Ekaterinburg) by photographing visual results of vandal acts (6217 photos) with subsequent trace-assessment and image content analysis, as well as diagnostics of personal characteristics and motivational basis of vandal activity of possible subjects of vandalism among youth. The results of the study allowed to identify the functions of vandalism at the socio-environmental and individual-subjective levels. The socio-environmental functions of vandalism include the signaling function, the function of preparing of social changes, the constructing function, and the function of managing public moods. The demonstrative-protest function, the response function, the refund function, and the self-expression function are assigned to the individual-subjective functions of vandalism. A two-dimensional model of vandal functions has been formed, where functions are distributed in the spaces 'construction reconstruction', 'emotional regulation/moral regulation'. It is noted that any function of vandal activity at the individual level becomes a kind of marker of 'points of tension' at the social and environmental level. Acknowledgment: The research was supported financially by Russian Science Foundation, (Project No. 17-18-01278).Keywords: destruction, urban environment, vandal behavior, vandalism, vandalism functions
Procedia PDF Downloads 20176 Vehicle Timing Motion Detection Based on Multi-Dimensional Dynamic Detection Network
Authors: Jia Li, Xing Wei, Yuchen Hong, Yang Lu
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Detecting vehicle behavior has always been the focus of intelligent transportation, but with the explosive growth of the number of vehicles and the complexity of the road environment, the vehicle behavior videos captured by traditional surveillance have been unable to satisfy the study of vehicle behavior. The traditional method of manually labeling vehicle behavior is too time-consuming and labor-intensive, but the existing object detection and tracking algorithms have poor practicability and low behavioral location detection rate. This paper proposes a vehicle behavior detection algorithm based on the dual-stream convolution network and the multi-dimensional video dynamic detection network. In the videos, the straight-line behavior of the vehicle will default to the background behavior. The Changing lanes, turning and turning around are set as target behaviors. The purpose of this model is to automatically mark the target behavior of the vehicle from the untrimmed videos. First, the target behavior proposals in the long video are extracted through the dual-stream convolution network. The model uses a dual-stream convolutional network to generate a one-dimensional action score waveform, and then extract segments with scores above a given threshold M into preliminary vehicle behavior proposals. Second, the preliminary proposals are pruned and identified using the multi-dimensional video dynamic detection network. Referring to the hierarchical reinforcement learning, the multi-dimensional network includes a Timer module and a Spacer module, where the Timer module mines time information in the video stream and the Spacer module extracts spatial information in the video frame. The Timer and Spacer module are implemented by Long Short-Term Memory (LSTM) and start from an all-zero hidden state. The Timer module uses the Transformer mechanism to extract timing information from the video stream and extract features by linear mapping and other methods. Finally, the model fuses time information and spatial information and obtains the location and category of the behavior through the softmax layer. This paper uses recall and precision to measure the performance of the model. Extensive experiments show that based on the dataset of this paper, the proposed model has obvious advantages compared with the existing state-of-the-art behavior detection algorithms. When the Time Intersection over Union (TIoU) threshold is 0.5, the Average-Precision (MP) reaches 36.3% (the MP of baselines is 21.5%). In summary, this paper proposes a vehicle behavior detection model based on multi-dimensional dynamic detection network. This paper introduces spatial information and temporal information to extract vehicle behaviors in long videos. Experiments show that the proposed algorithm is advanced and accurate in-vehicle timing behavior detection. In the future, the focus will be on simultaneously detecting the timing behavior of multiple vehicles in complex traffic scenes (such as a busy street) while ensuring accuracy.Keywords: vehicle behavior detection, convolutional neural network, long short-term memory, deep learning
Procedia PDF Downloads 13075 Silver-Doped Magnetite Titanium Oxide Nanoparticles for Photocatalytic Degradation of Organic Pollutants
Authors: Hanna Abbo, Siyasanga Noganta, Salam Titinchi
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The global lack of clean water for human sanitation and other purposes has become an emerging dilemma for human beings. The presence of organic pollutants in wastewater produced by textile industries, leather manufacturing and chemical industries is an alarming matter for a safe environment and human health. For the last decades, conventional methods have been applied for the purification of water but due to industrialization these methods fall short. Advanced oxidation processes and their reliable application in degradation of many contaminants have been reported as a potential method to reduce and/or alleviate this problem. Lately it has been assumed that incorporation of some metal nanoparticles such as magnetite nanoparticles as photocatalyst for Fenton reaction which could improve the degradation efficiency of contaminants. Core/shell nanoparticles, are extensively studied because of their wide applications in the biomedical, drug delivery, electronics fields and water treatment. The current study is centred on the synthesis of silver-doped Fe3O4/SiO2/TiO2 photocatalyst. Magnetically separable Fe3O4@SiO2@TiO2 composite with core–shell structure were synthesized by the deposition of uniform anatase TiO2 NPs on Fe3O4@SiO2 by using titanium butoxide (TBOT) as titanium source. Then, the silver is doped on SiO2 layer by hydrothermal method. Integration of magnetic nanoparticles was suggested to avoid the post separation difficulties associated with the powder form of the TiO2 catalyst, increase of the surface area and adsorption properties. The morphology, structure, composition, and magnetism of the resulting composites were characterized and their photocatalytic activities were also evaluated. The results demonstrate that TiO2 NPs were uniformly deposited on the Fe3O4@SiO2 surface. The silver nanoparticles were also uniformly distributed on the surface of TiO2 nanoparticles. The aim of this work is to study the suitability of photocatalysis for the treatment of aqueous streams containing organic pollutants such as methylene blue which is selected as a model compound to represent one of the pollutants existing in wastewaters. Various factors such as initial pollutant concentration, photocatalyst dose and wastewater matrix were studied for their effect on the photocatalytic degradation of the organic model pollutants using the as synthesized catalysts and compared with the commercial titanium dioxide (Aeroxide P25). Photocatalysis was found to be a potential purification method for the studied pollutant also in an industrial wastewater matrix with the removal percentages of over 81 % within 15 minutes. Methylene blue was removed most efficiently and its removal consumed the least of energy in terms of the specific applied energy. The magnetic Ag/SiO2/TiO2 composites show high photocatalytic performance and can be recycled three times by magnetic separation without major loss of activity, which meant that they can be used as efficient and conveniently renewable photocatalyst.Keywords: Magnetite nanoparticles, Titanium, Photocatalyst, Organic pollutant, Water treatment
Procedia PDF Downloads 26774 Carbon-Foam Supported Electrocatalysts for Polymer Electrolyte Membrane Fuel Cells
Authors: Albert Mufundirwa, Satoru Yoshioka, K. Ogi, Takeharu Sugiyama, George F. Harrington, Bretislav Smid, Benjamin Cunning, Kazunari Sasaki, Akari Hayashi, Stephen M. Lyth
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Polymer electrolyte membrane fuel cells (PEMFCs) are electrochemical energy conversion devices used for portable, residential and vehicular applications due to their low emissions, high efficiency, and quick start-up characteristics. However, PEMFCs generally use expensive, Pt-based electrocatalysts as electrode catalysts. Due to the high cost and limited availability of platinum, research and development to either drastically reduce platinum loading, or replace platinum with alternative catalysts is of paramount importance. A combination of high surface area supports and nano-structured active sites is essential for effective operation of catalysts. We synthesize carbon foam supports by thermal decomposition of sodium ethoxide, using a template-free, gram scale, cheap, and scalable pyrolysis method. This carbon foam has a high surface area, highly porous, three-dimensional framework which is ideal for electrochemical applications. These carbon foams can have surface area larger than 2500 m²/g, and electron microscopy reveals that they have micron-scale cells, separated by few-layer graphene-like carbon walls. We applied this carbon foam as a platinum catalyst support, resulting in the improved electrochemical surface area and mass activity for the oxygen reduction reaction (ORR), compared to carbon black. Similarly, silver-decorated carbon foams showed higher activity and efficiency for electrochemical carbon dioxide conversion than silver-decorated carbon black. A promising alternative to Pt-catalysts for the ORR is iron-impregnated nitrogen-doped carbon catalysts (Fe-N-C). Doping carbon with nitrogen alters the chemical structure and modulates the electronic properties, allowing a degree of control over the catalytic properties. We have adapted our synthesis method to produce nitrogen-doped carbon foams with large surface area, using triethanolamine as a nitrogen feedstock, in a novel bottom-up protocol. These foams are then infiltrated with iron acetate (FeAc) and pyrolysed to form Fe-N-C foams. The resulting Fe-N-C foam catalysts have high initial activity (half-wave potential of 0.68 VRHE), comparable to that of commercially available Pt-free catalysts (e.g., NPC-2000, Pajarito Powder) in acid solution. In alkaline solution, the Fe-N-C carbon foam catalysts have a half-wave potential of 0.89 VRHE, which is higher than that of NPC-2000 by almost 10 mVRHE, and far out-performing platinum. However, the durability is still a problem at present. The lessons learned from X-ray absorption spectroscopy (XAS), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), and electrochemical measurements will be used to carefully design Fe-N-C catalysts for higher performance PEMFCs.Keywords: carbon-foam, polymer electrolyte membrane fuel cells, platinum, Pt-free, Fe-N-C, ORR
Procedia PDF Downloads 18073 Spatial Organization of Cells over the Process of Pellicle Formation by Pseudomonas alkylphenolica KL28
Authors: Kyoung Lee
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Numerous aerobic bacteria have the ability to form multicellular communities on the surface layer of the air-liquid (A-L) interface as a biofilm called a pellicle. Pellicles occupied at the A-L interface will benefit from the utilization of oxygen from air and nutrient from liquid. Buoyancy of cells can be obtained by high surface tension at the A-L interface. Thus, formation of pellicles is an adaptive advantage in utilization of excess nutrients in the standing culture where oxygen depletion is easily set up due to rapid cell growth. In natural environments, pellicles are commonly observed on the surface of lake or pond contaminated with pollutants. Previously, we have shown that when cultured in standing LB media an alkylphenol-degrading bacteria Pseudomonas alkylphenolia KL28 forms pellicles in a diameter of 0.3-0.5 mm with a thickness of ca 40 µm. The pellicles have unique features for possessing flatness and unusual rigidity. In this study, the biogenesis of the circular pellicles has been investigated by observing the cell organization at early stages of pellicle formation and cell arrangements in pellicle, providing a clue for highly organized cellular arrangement to be adapted to the air-liquid niche. Here, we first monitored developmental patterns of pellicle from monolayer to multicellular organization. Pellicles were shaped by controlled growth of constituent cells which accumulate extracellular polymeric substance. The initial two-dimensional growth was transited to multilayers by a constraint force of accumulated self-produced extracellular polymeric substance. Experiments showed that pellicles are formed by clonal growth and even with knock-out of genes for flagella and pilus formation. In contrast, the mutants in the epm gene cluster for alginate-like polymer biosynthesis were incompetent in cell alignment for initial two-dimensional growth of pellicles. Electron microscopic and confocal laser scanning microscopic studies showed that the fully matured structures are highly packed by matrix-encased cells which have special arrangements. The cells on the surface of the pellicle lie relatively flat and inside longitudinally cross packed. HPLC analysis of the extrapolysaccharide (EPS) hydrolysate from the colonies from LB agar showed a composition with L-fucose, L-rhamnose, D-galactosamine, D-glucosamine, D-galactose, D-glucose, D-mannose. However, that from pellicles showed similar neutral and amino sugar profile but missing galactose. Furthermore, uronic acid analysis of EPS hydrolysates by HPLC showed that mannuronic acid was detected from pellicles not from colonies, indicating the epm-derived polymer is critical for pellicle formation as proved by the epm mutants. This study verified that for the circular pellicle architecture P. alkylphenolica KL28 cells utilized EPS building blocks different from that used for colony construction. These results indicate that P. alkylphenolica KL28 is a clever architect that dictates unique cell arrangements with selected EPS matrix material to construct sophisticated building, circular biofilm pellicles.Keywords: biofilm, matrix, pellicle, pseudomonas
Procedia PDF Downloads 15372 Purple Spots on Historical Parchments: Confirming the Microbial Succession at the Basis of Biodeterioration
Authors: N. Perini, M. C. Thaller, F. Mercuri, S. Orlanducci, A. Rubechini, L. Migliore
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The preservation of cultural heritage is one of the major challenges of today’s society, because of the fundamental right of future generations to inherit it as the continuity with their historical and cultural identity. Parchments, consisting of a semi-solid matrix of collagen produced from animal skin (i.e., sheep or goats), are a significant part of the cultural heritage, being used as writing material for many centuries. Due to their animal origin, parchments easily undergo biodeterioration. The most common biological damage is characterized by isolated or coalescent purple spots that often leads to the detachment of the superficial layer and the loss of the written historical content of the document. Although many parchments with the same biodegradative features were analyzed, no common causative agent has been found so far. Very recently, a study was performed on a purple-damaged parchment roll dated back 1244 A.D, the A.A. Arm. I-XVIII 3328, belonging to the oldest collection of the Vatican Secret Archive (Fondo 'Archivum Arcis'), by comparing uncolored undamaged and purple damaged areas of the same document. As a whole, the study gave interesting results to hypothesize a model of biodeterioration, consisting of a microbial succession acting in two main phases: the first one, common to all the damaged parchments, is characterized by halophilic and halotolerant bacteria fostered by the salty environment within the parchment maybe induced by bringing of the hides; the second one, changing with the individual history of each parchment, determines the identity of its colonizers. The design of this model was pivotal to this study, performed by different labs of the Tor Vergata University (Rome, Italy), in collaboration with the Vatican Secret Archive. Three documents, belonging to a collection of dramatically damaged parchments archived as 'Faldone Patrizi A 19' (dated back XVII century A.D.), were analyzed through a multidisciplinary approach, including three updated technologies: (i) Next Generation Sequencing (NGS, Illumina) to describe the microbial communities colonizing the damaged and undamaged areas, (ii) RAMAN spectroscopy to analyze the purple pigments, (iii) Light Transmitted Analysis (LTA) to evaluate the kind and entity of the damage to native collagen. The metagenomic analysis obtained from NGS revealed DNA sequences belonging to Halobacterium salinarum mainly in the undamaged areas. RAMAN spectroscopy detected pigments within the purple spots, mainly bacteriorhodopsine/rhodopsin-like pigments, a purple transmembrane protein containing retinal and present in Halobacteria. The LTA technique revealed extremely damaged collagen structures in both damaged and undamaged areas of the parchments. In the light of these data, the study represents a first confirmation of the microbial succession model described above. The demonstration of this model is pivotal to start any possible new restoration strategy to bring back historical parchments to their original beauty, but also to open opportunities for intervention on a huge amount of documents.Keywords: biodeterioration, parchments, purple spots, ecological succession
Procedia PDF Downloads 17171 Heat Accumulation in Soils of Belarus
Authors: Maryna Barushka, Aleh Meshyk
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The research analyzes absolute maximum soil temperatures registered at 36 gauge stations in Belarus from 1950 to 2013. The main method applied in the research is cartographic, in particular, trend surface analysis. Warming that had never been so long and intensive before started in 1988. The average temperature in January and February of that year exceeded the norm by 7-7.5 С, in March and April by 3-5С. In general, that year, as well as the year of 2008, happened to be the hottest ones in the whole period of instrumental observation. Yearly average air temperature in Belarus in those years was +8.0-8.2 С, which exceeded the norm by 2.0 – 2.2 С. The warming has been observed so far. The only exception was in 1996 when the yearly average air temperature in Belarus was below normal by 0.5 С. In Belarus the value of trend line of standard temperature deviation in the warmest months (July-August) has been positive for the past 25 years. In 2010 absolute maximum air and soil temperature exceeded the norm at 15 gauge stations in Belarus. The structure of natural processes includes global, regional, and local constituents. Trend surface analysis of the investigated characteristics makes it possible to determine global, regional, and local components. Linear trend surface shows the occurrence of weather deviations on a global scale, outside Belarus. Maximum soil temperature appears to be growing in the south-west direction with the gradient of 5.0 С. It is explained by the latitude factor. Polynomial trend surfaces show regional peculiarities of Belarus. Extreme temperature regime is formed due to some factors. The prevailing one is advection of turbulent flow of the ground layer of the atmosphere. In summer influence of the Azores High producing anticyclones is great. The Gulf Stream current forms the values of temperature trends in a year period. The most intensive flow of the Gulf Stream in the second half of winter and the second half of summer coincides with the periods of maximum temperature trends in Belarus. It is possible to estimate a local component of weather deviations in the analysis of the difference in values of the investigated characteristics and their trend surfaces. Maximum positive deviation (up to +4 С) of averaged soil temperature corresponds to the flat terrain in Pripyat Polesie, Brest Polesie, and Belarusian Poozerie Area. Negative differences correspond to the higher relief which partially compensates extreme heat regime of soils. Another important factor for maximum soil temperature in these areas is peat-bog soils with the least albedo of 8-15%. As yearly maximum soil temperature reaches 40-60 С, this could be both negative and positive factors for Belarus’s environment and economy. High temperature causes droughts resulting in crops dying and soil blowing. On the other hand, vegetation period has lengthened thanks to bigger heat resources, which allows planting such heat-loving crops as melons and grapes with appropriate irrigation. Thus, trend surface analysis allows determining global, regional, and local factors in accumulating heat in the soils of Belarus.Keywords: soil, temperature, trend surface analysis, warming
Procedia PDF Downloads 13470 Hydrogeological Appraisal of Karacahisar Coal Field (Western Turkey): Impacts of Mining on Groundwater Resources Utilized for Water Supply
Authors: Sukran Acikel, Mehmet Ekmekci, Otgonbayar Namkhai
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Lignite coal fields in western Turkey generally occurs in tensional Neogene basins bordered by major faults. Karacahisar coal field in Mugla province of western Turkey is a large Neogene basin filled with alternation of silisic and calcerous layers. The basement of the basin is composed of mainly karstified carbonate rocks of Mesozoic and schists of Paleozoic age. The basement rocks are exposed at highlands surrounding the basin. The basin fill deposits forms shallow, low yield and local aquifers whereas karstic carbonate rock masses forms the major aquifer in the region. The karstic aquifer discharges through a spring zone issuing at intersection of two major faults. Municipal water demand in Bodrum city, a touristic attraction area is almost totally supplied by boreholes tapping the karstic aquifer. A well field has been constructed on the eastern edge of the coal basin, which forms a ridge separating two Neogene basins. A major concern was raised about the plausible impact of mining activities on groundwater system in general and on water supply well field in particular. The hydrogeological studies carried out in the area revealed that the coal seam is located below the groundwater level. Mining operations will be affected by groundwater inflow to the pits, which will require dewatering measures. Dewatering activities in mine sites have two-sided effects: a) lowers the groundwater level at and around the pit for a safe and effective mining operation, b) continuous dewatering causes expansion of cone of depression to reach a spring, stream and/or well being utilized by local people, capturing their water. Plausible effect of mining operations on the flow of the spring zone was another issue of concern. Therefore, a detailed representative hydrogeological conceptual model of the site was developed on the basis of available data and field work. According to the hydrogeological conceptual model, dewatering of Neogene layers will not hydraulically affect the water supply wells, however, the ultimate perimeter of the open pit will expand to intersect the well field. According to the conceptual model, the coal seam is separated from the bottom by a thick impervious clay layer sitting on the carbonate basement. Therefore, the hydrostratigraphy does not allow a hydraulic interaction between the mine pit and the karstic carbonate rock aquifer. However, the structural setting in the basin suggests that deep faults intersecting the basement and the Neogene sequence will most probably carry the deep groundwater up to a level above the bottom of the pit. This will require taking necessary measure to lower the piezometric level of the carbonate rock aquifer along the faults. Dewatering the carbonate rock aquifer will reduce the flow to the spring zone. All findings were put together to recommend a strategy for safe and effective mining operation.Keywords: conceptual model, dewatering, groundwater, mining operation
Procedia PDF Downloads 40069 Sustainability in Space: Implementation of Circular Economy and Material Efficiency Strategies in Space Missions
Authors: Hamda M. Al-Ali
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The ultimate aim of space exploration has been centralized around the possibility of life on other planets in the solar system. This aim is driven by the detrimental effects that climate change could potentially have on human survival on Earth in the future. This drives humans to search for feasible solutions to increase environmental and economical sustainability on Earth and to evaluate and explore the ability of human survival on other planets such as Mars. To do that, frequent space missions are required to meet the ambitious human goals. This means that reliable and affordable access to space is required, which could be largely achieved through the use of reusable spacecrafts. Therefore, materials and resources must be used wisely to meet the increasing demand. Space missions are currently extremely expensive to operate. However, reusing materials hence spacecrafts, can potentially reduce overall mission costs as well as the negative impact on both space and Earth environments. This is because reusing materials leads to less waste generated per mission, and therefore fewer landfill sites are required. Reusing materials reduces resource consumption, material production, and the need for processing new and replacement spacecraft and launch vehicle parts. Consequently, this will ease and facilitate human access to outer space as it will reduce the demand for scarce resources, which will boost material efficiency in the space industry. Material efficiency expresses the extent to which resources are consumed in the production cycle and how the waste produced by the industrial process is minimized. The strategies proposed in this paper to boost material efficiency in the space sector are the introduction of key performance indicators that are able to measure material efficiency as well as the introduction of clearly defined policies and legislation that can be easily implemented within the general practices in the space industry. Another strategy to improve material efficiency is by amplifying energy and resource efficiency through reusing materials. The circularity of various spacecraft materials such as Kevlar, steel, and aluminum alloys could be maximized through reusing them directly or after galvanizing them with another layer of material to act as a protective coat. This research paper has an aim to investigate and discuss how to improve material efficiency in space missions considering circular economy concepts so that space and Earth become more economically and environmentally sustainable. The circular economy is a transition from a make-use-waste linear model to a closed-loop socio-economic model, which is regenerative and restorative in nature. The implementation of a circular economy will reduce waste and pollution through maximizing material efficiency, ensuring that businesses can thrive and sustain. Further research into the extent to which reusable launch vehicles reduce space mission costs have been discussed, along with the environmental and economic implications it could have on the space sector and the environment. This has been examined through research and in-depth literature review of published reports, books, scientific articles, and journals. Keywords such as material efficiency, circular economy, reusable launch vehicles and spacecraft materials were used to search for relevant literature.Keywords: circular economy, key performance indicator, material efficiency, reusable launch vehicles, spacecraft materials
Procedia PDF Downloads 12568 NEOM Coast from Intertidal to Sabkha Systems: A Geological Overview
Authors: Mohamed Abouelresh, Subhajit Kumar, Lamidi Babalola, Septriandi Chan, Ali Al Musabeh A., Thadickal V. Joydas, Bruno Pulido
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Neom has a relatively long coastline on the Red Sea and the Gulf of Aqaba, which is about 300 kilometres long, in addition to many naturally formed bays along the Red Sea coast. Undoubtedly, these coasts provide an excellent opportunity for tourism and other activities; however, these coastal areas host a wide range of salinity-dependent ecosystems that need to be protected. The main objective of the study was to identify the coastal features, including tidal flats and salt flats, along the NEOM coast. A base map of the study area generated from the satellite images contained the main landform features and, in particular, the boundaries of the inland and coastal sabkhas. A field survey was conducted to map and characterize the intertidal and sabkha landforms. The coastal and inner coastal areas of NEOM are mainly covered by the quaternary sediments, which include gravel sheets, terraces, raised reef limestone, evaporite successions, eolian dunes, and undifferentiated sand/gravel deposits (alluvium, alluvial outwash, wind-blown sand beach). There are different landforms that characterizes the NEOM coast, including rocky coast, tidal zone, and sabkha. Sabkha area ranges between a few to tens of square kilometers. Coastal sabkha extended across the shoreline of NEOM, specifically at Gayal and Sharma areas, while the continental sabkha only existed at Gayal Town. The inland Sabkha at Gayal is mainly composed of a thin (15-25 cm) evaporite crust composed of a dark brown, cavernous, rugged, pitted, colloidal salty sand layer with salt-tolerant vegetation. The inland Sabkha is considered a groundwater-driven sedimentary system as indicated by syndepositional intra-sediment capillary evaporites, which precipitate in both marine and continental salt flats. Gayal coastal Sabkha is made up of tidal inlets, tidal creeks, and lagoons followed in a landward direction with well-developed sabkha layers. The surface sediments of the coastal Sabkha are composed of unlithified calcareous, gypsiferous, coarse to medium sands, and silt with bioclastic fragments underlain by several organic-rich layers. The coastal flat is graded landward into widespread, flat vegetated Sabkhas dissected by tributaries of the fluvial system, which debouches to the Red Sea. The coast from Gayal to Magna through Ras El-Sheikh Humaid is continuously subjected to tidal flows, which create an intertidal depositional system. The intertidal flats at NEOM are extensive, nearly horizontal land forming a very dynamic system in which several physical, chemical, geomorphological, and biological processes are acting simultaneously. The current work provides a field-based identification of the coastal sabkha and intertidal sites at NEOM. However, the mutual interaction between tidal flows and sabkha development, particularly at Gayal, needs to be well understood through comprehensive field and lab analysis.Keywords: coast, intertidal, deposition, sabkha
Procedia PDF Downloads 8467 Edible Active Antimicrobial Coatings onto Plastic-Based Laminates and Its Performance Assessment on the Shelf Life of Vacuum Packaged Beef Steaks
Authors: Andrey A. Tyuftin, David Clarke, Malco C. Cruz-Romero, Declan Bolton, Seamus Fanning, Shashi K. Pankaj, Carmen Bueno-Ferrer, Patrick J. Cullen, Joe P. Kerry
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Prolonging of shelf-life is essential in order to address issues such as; supplier demands across continents, economical profit, customer satisfaction, and reduction of food wastage. Smart packaging solutions presented in the form of naturally occurred antimicrobially-active packaging may be a solution to these and other issues. Gelatin film forming solution with adding of natural sourced antimicrobials is a promising tool for the active smart packaging. The objective of this study was to coat conventional plastic hydrophobic packaging material with hydrophilic antimicrobial active beef gelatin coating and conduct shelf life trials on beef sub-primal cuts. Minimal inhibition concentration (MIC) of Caprylic acid sodium salt (SO) and commercially available Auranta FV (AFV) (bitter oranges extract with mixture of nutritive organic acids) were found of 1 and 1.5 % respectively against bacterial strains Bacillus cereus, Pseudomonas fluorescens, Escherichia coli, Staphylococcus aureus and aerobic and anaerobic beef microflora. Therefore SO or AFV were incorporated in beef gelatin film forming solution in concentration of two times of MIC which was coated on a conventional plastic LDPE/PA film on the inner cold plasma treated polyethylene surface. Beef samples were vacuum packed in this material and stored under chilling conditions, sampled at weekly intervals during 42 days shelf life study. No significant differences (p < 0.05) in the cook loss was observed among the different treatments compared to control samples until the day 29. Only for AFV coated beef sample it was 3% higher (37.3%) than the control (34.4 %) on the day 36. It was found antimicrobial films did not protect beef against discoloration. SO containing packages significantly (p < 0.05) reduced Total viable bacterial counts (TVC) compared to the control and AFV samples until the day 35. No significant reduction in TVC was observed between SO and AFV films on the day 42 but a significant difference was observed compared to control samples with a 1.40 log of bacteria reduction on the day 42. AFV films significantly (p < 0.05) reduced TVC compared to control samples from the day 14 until the day 42. Control samples reached the set value of 7 log CFU/g on day 27 of testing, AFV films did not reach this set limit until day 35 and SO films until day 42 of testing. The antimicrobial AFV and SO coated films significantly prolonged the shelf-life of beef steaks by 33 or 55% (on 7 and 14 days respectively) compared to control film samples. It is concluded antimicrobial coated films were successfully developed by coating the inner polyethylene layer of conventional LDPE/PA laminated films after plasma surface treatment. The results indicated that the use of antimicrobial active packaging coated with SO or AFV increased significantly (p < 0.05) the shelf life of the beef sub-primal. Overall, AFV or SO containing gelatin coatings have the potential of being used as effective antimicrobials for active packaging applications for muscle-based food products.Keywords: active packaging, antimicrobials, edible coatings, food packaging, gelatin films, meat science
Procedia PDF Downloads 30366 Nuclear Powered UAV for Surveillances and Aerial Photography
Authors: Rajasekar Elangopandian, Anand Shanmugam
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Now-a-days for surveillances unmanned aerial vehicle plays a vital role. Not only for surveillances, aerial photography disaster management and the notice of earth behavior UAV1s envisages meticulously. To reduce the maintenance and fuel nuclear powered Vehicles are greater support. The design consideration is much important for the UAV manufacturing industry and Research and development agency. Eventually design is looking like a pentagon shaped fuselage and black rubber coated paint in order to escape from the enemy radar and other targets. The pentagon shape fuselage has large space to keep the mini nuclear reactor inside and the material is carbon – carbon fiber specially designed by the software called cosmol and hyper mesh 14.2. So the weight consideration will produce the positive result for productivity. The walls of the fuselage are coated with lead and protective shield. A double layer of W/Bi sheet is proposed for radiation protection at the energy range of 70 Kev to 90 Kev. The designed W/bi sheet, only 0.14 mm thick and is 36% light. The properties of the fillers were determined from zeta potential and particle size measurements. The Exposes of the radiation can be attenuated by 3 ways such as minimizing exposure time, Maximizing distance from the radiation source and shielding the whole vehicle. The inside reactor will be switched ON when the UAV starts its cruise. The moderators and the control rods can be inserted by automation technique by newly developed software. The heat generated by the reactor will be used to run the turbine which is fixed inside the UAV called mini turbine with natural rubber composite Shaft radiation shield. Cooling system will be in two mode such as liquid and air cooled. Liquid coolant for the heat regeneration is ordinary water, liquid sodium, helium and the walls are made up of regenerative and radiation protective material. The other components like camera and arms bay will be located at the bottom of the UAV high are specially made products in order to escape from the radiation. They are coated with lead Pb and natural rubber composite material. This technique provides the long rang and endurance for eternal flight mission until we need any changeability of parts or product. This UAV has the special advantage of ` land on String` means it`ll land at electric line to charge the automated electronics. Then the fuel is enriched uranium (< 5% U - 235) contains hundreds of fuel pins. This technique provides eternal duty for surveillances and aerial photography. The landing of the vehicle is ease of operation likewise the takeoff is also easier than any other mechanism which present in nowadays. This UAV gives great immense and immaculate technology for surveillance and target detecting and smashing the target.Keywords: mini turbine, liquid coolant for the heat regeneration, in order to escape from the radiation, eternal flight mission, it`ll land at electric line
Procedia PDF Downloads 41065 Biodegradable Cross-Linked Composite Hydrogels Enriched with Small Molecule for Osteochondral Regeneration
Authors: Elena I. Oprita, Oana Craciunescu, Rodica Tatia, Teodora Ciucan, Reka Barabas, Orsolya Raduly, Anca Oancea
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Healing of osteochondral defects requires repair of the damaged articular cartilage, the underlying subchondral bone and the interface between these tissues (the functional calcified layer). For this purpose, developing a single monophasic scaffold that can regenerate two specific lineages (cartilage and bone) becomes a challenge. The aim of this work was to develop variants of biodegradable cross-linked composite hydrogel based on natural polypeptides (gelatin), polysaccharides components (chondroitin-4-sulphate and hyaluronic acid), in a ratio of 2:0.08:0.02 (w/w/w) and mixed with Si-hydroxyapatite (Si-Hap), in two ratios of 1:1 and 2:1 (w/w). Si-Hap was synthesized and characterized as a better alternative to conventional Hap. Subsequently, both composite hydrogel variants were cross-linked with (N, N-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC) and enriched with a small bioactive molecule (icariin). The small molecule icariin (Ica) (C33H40O15) is the main active constituent (flavonoid) of Herba epimedium used in traditional Chinese medicine to cure bone- and cartilage-related disorders. Ica enhances osteogenic and chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), facilitates matrix calcification and increases the specific extracellular matrix (ECM) components synthesis by chondrocytes. Afterward, the composite hydrogels were characterized for their physicochemical properties in terms of the enzymatic biodegradation in the presence of type I collagenase and trypsin, the swelling capacity and the degree of crosslinking (TNBS assay). The cumulative release of Ica and real-time concentration were quantified at predetermined periods of time, according to the standard curve of standard Ica, after hydrogels incubation in saline buffer at physiological parameters. The obtained cross-linked composite hydrogels enriched with small-molecule Ica were also characterized for morphology by scanning electron microscopy (SEM). Their cytocompatibility was evaluated according to EN ISO 10993-5:2009 standard for medical device testing. Thus, analyses regarding cell viability (Live/Dead assay), cell proliferation (Neutral Red assay) and cell adhesion to composite hydrogels (SEM) were performed using NCTC clone L929 cell line. The final results showed that both cross-linked composite hydrogel variants enriched with Ica presented optimal physicochemical, structural and biological properties to be used as a natural scaffold able to repair osteochondral defects. The data did not reveal any toxicity of composite hydrogels in NCTC stabilized cell lines within the tested range of concentrations. Moreover, cells were capable of spreading and proliferating on both composite hydrogel surfaces. In conclusion, the designed biodegradable cross-linked composites enriched with Si and Ica are recommended for further testing as natural temporary scaffolds, which can allow cell migration and synthesis of new extracellular matrix within osteochondral defects.Keywords: composites, gelatin, osteochondral defect, small molecule
Procedia PDF Downloads 17564 Convolutional Neural Network Based on Random Kernels for Analyzing Visual Imagery
Authors: Ja-Keoung Koo, Kensuke Nakamura, Hyohun Kim, Dongwha Shin, Yeonseok Kim, Ji-Su Ahn, Byung-Woo Hong
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The machine learning techniques based on a convolutional neural network (CNN) have been actively developed and successfully applied to a variety of image analysis tasks including reconstruction, noise reduction, resolution enhancement, segmentation, motion estimation, object recognition. The classical visual information processing that ranges from low level tasks to high level ones has been widely developed in the deep learning framework. It is generally considered as a challenging problem to derive visual interpretation from high dimensional imagery data. A CNN is a class of feed-forward artificial neural network that usually consists of deep layers the connections of which are established by a series of non-linear operations. The CNN architecture is known to be shift invariant due to its shared weights and translation invariance characteristics. However, it is often computationally intractable to optimize the network in particular with a large number of convolution layers due to a large number of unknowns to be optimized with respect to the training set that is generally required to be large enough to effectively generalize the model under consideration. It is also necessary to limit the size of convolution kernels due to the computational expense despite of the recent development of effective parallel processing machinery, which leads to the use of the constantly small size of the convolution kernels throughout the deep CNN architecture. However, it is often desired to consider different scales in the analysis of visual features at different layers in the network. Thus, we propose a CNN model where different sizes of the convolution kernels are applied at each layer based on the random projection. We apply random filters with varying sizes and associate the filter responses with scalar weights that correspond to the standard deviation of the random filters. We are allowed to use large number of random filters with the cost of one scalar unknown for each filter. The computational cost in the back-propagation procedure does not increase with the larger size of the filters even though the additional computational cost is required in the computation of convolution in the feed-forward procedure. The use of random kernels with varying sizes allows to effectively analyze image features at multiple scales leading to a better generalization. The robustness and effectiveness of the proposed CNN based on random kernels are demonstrated by numerical experiments where the quantitative comparison of the well-known CNN architectures and our models that simply replace the convolution kernels with the random filters is performed. The experimental results indicate that our model achieves better performance with less number of unknown weights. The proposed algorithm has a high potential in the application of a variety of visual tasks based on the CNN framework. Acknowledgement—This work was supported by the MISP (Ministry of Science and ICT), Korea, under the National Program for Excellence in SW (20170001000011001) supervised by IITP, and NRF-2014R1A2A1A11051941, NRF2017R1A2B4006023.Keywords: deep learning, convolutional neural network, random kernel, random projection, dimensionality reduction, object recognition
Procedia PDF Downloads 29063 Modeling Taxane-Induced Peripheral Neuropathy Ex Vivo Using Patient-Derived Neurons
Authors: G. Cunningham, E. Cantor, X. Wu, F. Shen, G. Jiang, S. Philips, C. Bales, Y. Xiao, T. R. Cummins, J. C. Fehrenbacher, B. P. Schneider
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Background: Taxane-induced peripheral neuropathy (TIPN) is the most devastating survivorship issue for patients receiving therapy. Dose reductions due to TIPN in the curative setting lead to inferior outcomes for African American patients, as prior research has shown that this group is more susceptible to developing severe neuropathy. The mechanistic underpinnings of TIPN, however, have not been entirely elucidated. While it would be appealing to use primary tissue to study the development of TIPN, procuring nerves from patients is not realistically feasible, as nerve biopsies are painful and may result in permanent damage. Therefore, our laboratory has investigated paclitaxel-induced neuronal morphological and molecular changes using an ex vivo model of human-induced pluripotent stem cell (iPSC)-derived neurons. Methods: iPSCs are undifferentiated and endlessly dividing cells that can be generated from a patient’s somatic cells, such as peripheral blood mononuclear cells (PBMCs). We successfully reprogrammed PBMCs into iPSCs using the Erythroid Progenitor Reprograming Kit (STEMCell Technologiesᵀᴹ); pluripotency was verified by flow cytometry analysis. iPSCs were then induced into neurons using a differentiation protocol that bypasses the neural progenitor stage and uses selected small-molecule modulators of key signaling pathways (SMAD, Notch, FGFR1 inhibition, and Wnt activation). Results: Flow cytometry analysis revealed expression of core pluripotency transcription factors Nanog, Oct3/4 and Sox2 in iPSCs overlaps with commercially purchased pluripotent cell line UCSD064i-20-2. Trilineage differentiation of iPSCs was confirmed with immunofluorescent imaging with germ-layer-specific markers; Sox17 and ExoA2 for ectoderm, Nestin, and Pax6 for mesoderm, and Ncam and Brachyury for endoderm. Sensory neuron markers, β-III tubulin, and Peripherin were applied to stain the cells for the maturity of iPSC-derived neurons. Patch-clamp electrophysiology and calcitonin gene-related peptide (CGRP) release data supported the functionality of the induced neurons and provided insight into the timing for which downstream assays could be performed (week 4 post-induction). We have also performed a cell viability assay and fluorescence-activated cell sorting (FACS) using four cell-surface markers (CD184, CD44, CD15, and CD24) to select a neuronal population. At least 70% of the cells were viable in the isolated neuron population. Conclusion: We have found that these iPSC-derived neurons recapitulate mature neuronal phenotypes and demonstrate functionality. Thus, this represents a patient-derived ex vivo neuronal model to investigate the molecular mechanisms of clinical TIPN.Keywords: chemotherapy, iPSC-derived neurons, peripheral neuropathy, taxane, paclitaxel
Procedia PDF Downloads 12262 Production of Medicinal Bio-active Amino Acid Gamma-Aminobutyric Acid In Dairy Sludge Medium
Authors: Farideh Tabatabaee Yazdi, Fereshteh Falah, Alireza Vasiee
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Introduction: Gamma-aminobutyric acid (GABA) is a non-protein amino acid that is widely present in organisms. GABA is a kind of pharmacological and biological component and its application is wide and useful. Several important physiological functions of GABA have been characterized, such as neurotransmission and induction of hypotension. GABA is also a strong secretagogue of insulin from the pancreas and effectively inhibits small airway-derived lung adenocarcinoma and tranquilizer. Many microorganisms can produce GABA, and lactic acid bacteria have been a focus of research in recent years because lactic acid bacteria possess special physiological activities and are generally regarded as safe. Among them, the Lb. Brevis produced the highest amount of GABA. The major factors affecting GABA production have been characterized, including carbon sources and glutamate concentration. The use of food industry waste to produce valuable products such as amino acids seems to be a good way to reduce production costs and prevent the waste of food resources. In a dairy factory, a high volume of sludge is produced from a separator that contains useful compounds such as growth factors, carbon, nitrogen, and organic matter that can be used by different microorganisms such as Lb.brevis as carbon and nitrogen sources. Therefore, it is a good source of GABA production. GABA is primarily formed by the irreversible α-decarboxylation reaction of L-glutamic acid or its salts, catalysed by the GAD enzyme. In the present study, this aim was achieved for the fast-growing of Lb.brevis and producing GABA, using the dairy industry sludge as a suitable growth medium. Lactobacillus Brevis strains obtained from Microbial Type Culture Collection (MTCC) were used as model strains. In order to prepare dairy sludge as a medium, sterilization should be done at 121 ° C for 15 minutes. Lb. Brevis was inoculated to the sludge media at pH=6 and incubated for 120 hours at 30 ° C. After fermentation, the supernatant solution is centrifuged and then, the GABA produced was analyzed by the Thin Layer chromatography (TLC) method qualitatively and by the high-performance liquid chromatography (HPLC) method quantitatively. By increasing the percentage of dairy sludge in the culture medium, the amount of GABA increased. Also, evaluated the growth of bacteria in this medium showed the positive effect of dairy sludge on the growth of Lb.brevis, which resulted in the production of more GABA. GABA-producing LAB offers the opportunity of developing naturally fermented health-oriented products. Although some GABA-producing LAB has been isolated to find strains suitable for different fermentations, further screening of various GABA-producing strains from LAB, especially high-yielding strains, is necessary. The production of lactic acid, bacterial gamma-aminobutyric acid, is safe and eco-friendly. The use of dairy industry waste causes enhanced environmental safety. Also provides the possibility of producing valuable compounds such as GABA. In general, dairy sludge is a suitable medium for the growth of Lactic Acid Bacteria and produce this amino acid that can reduce the final cost of it by providing carbon and nitrogen source.Keywords: GABA, Lactobacillus, HPLC, dairy sludge
Procedia PDF Downloads 14461 Biomimicked Nano-Structured Coating Elaboration by Soft Chemistry Route for Self-Cleaning and Antibacterial Uses
Authors: Elodie Niemiec, Philippe Champagne, Jean-Francois Blach, Philippe Moreau, Anthony Thuault, Arnaud Tricoteaux
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Hygiene of equipment in contact with users is an important issue in the railroad industry. The numerous cleanings to eliminate bacteria and dirt cost a lot. Besides, mechanical solicitations on contact parts are observed daily. It should be interesting to elaborate on a self-cleaning and antibacterial coating with sufficient adhesion and good resistance against mechanical and chemical solicitations. Thus, a Hauts-de-France and Maubeuge Val-de-Sambre conurbation authority co-financed Ph.D. thesis has been set up since October 2017 based on anterior studies carried by the Laboratory of Ceramic Materials and Processing. To accomplish this task, a soft chemical route has been implemented to bring a lotus effect on metallic substrates. It involves nanometric liquid zinc oxide synthesis under 100°C. The originality here consists in a variation of surface texturing by modification of the synthesis time of the species in solution. This helps to adjust wettability. Nanostructured zinc oxide has been chosen because of the inherent photocatalytic effect, which can activate organic substance degradation. Two methods of heating have been compared: conventional and microwave assistance. Tested subtracts are made of stainless steel to conform to transport uses. Substrate preparation was the first step of this protocol: a meticulous cleaning of the samples is applied. The main goal of the elaboration protocol is to fix enough zinc-based seeds to make them grow during the next step as desired (nanorod shaped). To improve this adhesion, a silica gel has been formulated and optimized to ensure chemical bonding between substrate and zinc seeds. The last step consists of deposing a wide carbonated organosilane to improve the superhydrophobic property of the coating. The quasi-proportionality between the reaction time and the nanorod length will be demonstrated. Water Contact (superior to 150°) and Roll-off Angle at different steps of the process will be presented. The antibacterial effect has been proved with Escherichia Coli, Staphylococcus Aureus, and Bacillus Subtilis. The mortality rate is found to be four times superior to a non-treated substrate. Photocatalytic experiences were carried out from different dyed solutions in contact with treated samples under UV irradiation. Spectroscopic measurements allow to determinate times of degradation according to the zinc quantity available on the surface. The final coating obtained is, therefore, not a monolayer but rather a set of amorphous/crystalline/amorphous layers that have been characterized by spectroscopic ellipsometry. We will show that the thickness of the nanostructured oxide layer depends essentially on the synthesis time set in the hydrothermal growth step. A green, easy-to-process and control coating with self-cleaning and antibacterial properties has been synthesized with a satisfying surface structuration.Keywords: antibacterial, biomimetism, soft-chemistry, zinc oxide
Procedia PDF Downloads 14360 The Monitor for Neutron Dose in Hadrontherapy Project: Secondary Neutron Measurement in Particle Therapy
Authors: V. Giacometti, R. Mirabelli, V. Patera, D. Pinci, A. Sarti, A. Sciubba, G. Traini, M. Marafini
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The particle therapy (PT) is a very modern technique of non invasive radiotherapy mainly devoted to the treatment of tumours untreatable with surgery or conventional radiotherapy, because localised closely to organ at risk (OaR). Nowadays, PT is available in about 55 centres in the word and only the 20\% of them are able to treat with carbon ion beam. However, the efficiency of the ion-beam treatments is so impressive that many new centres are in construction. The interest in this powerful technology lies to the main characteristic of PT: the high irradiation precision and conformity of the dose released to the tumour with the simultaneous preservation of the adjacent healthy tissue. However, the beam interactions with the patient produce a large component of secondary particles whose additional dose has to be taken into account during the definition of the treatment planning. Despite, the largest fraction of the dose is released to the tumour volume, a non-negligible amount is deposed in other body regions, mainly due to the scattering and nuclear interactions of the neutrons within the patient body. One of the main concerns in PT treatments is the possible occurrence of secondary malignant neoplasm (SMN). While SMNs can be developed up to decades after the treatments, their incidence impacts directly life quality of the cancer survivors, in particular in pediatric patients. Dedicated Treatment Planning Systems (TPS) are used to predict the normal tissue toxicity including the risk of late complications induced by the additional dose released by secondary neutrons. However, no precise measurement of secondary neutrons flux is available, as well as their energy and angular distributions: an accurate characterization is needed in order to improve TPS and reduce safety margins. The project MONDO (MOnitor for Neutron Dose in hadrOntherapy) is devoted to the construction of a secondary neutron tracker tailored to the characterization of that secondary neutron component. The detector, based on the tracking of the recoil protons produced in double-elastic scattering interactions, is a matrix of thin scintillating fibres, arranged in layer x-y oriented. The final size of the object is 10 x 10 x 20 cm3 (squared 250µm scint. fibres, double cladding). The readout of the fibres is carried out with a dedicated SPAD Array Sensor (SBAM) realised in CMOS technology by FBK (Fondazione Bruno Kessler). The detector is under development as well as the SBAM sensor and it is expected to be fully constructed for the end of the year. MONDO will make data tacking campaigns at the TIFPA Proton Therapy Center of Trento, at the CNAO (Pavia) and at HIT (Heidelberg) with carbon ion in order to characterize the neutron component and predict the additional dose delivered on the patients with much more precision and to drastically reduce the actual safety margins. Preliminary measurements with charged particles beams and MonteCarlo FLUKA simulation will be presented.Keywords: secondary neutrons, particle therapy, tracking detector, elastic scattering
Procedia PDF Downloads 22359 Piezotronic Effect on Electrical Characteristics of Zinc Oxide Varistors
Authors: Nadine Raidl, Benjamin Kaufmann, Michael Hofstätter, Peter Supancic
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If polycrystalline ZnO is properly doped and sintered under very specific conditions, it shows unique electrical properties, which are indispensable for today’s electronic industries, where it is used as the number one overvoltage protection material. Under a critical voltage, the polycrystalline bulk exhibits high electrical resistance but becomes suddenly up to twelve magnitudes more conductive if this voltage limit is exceeded (i.e., varistor effect). It is known that these peerless properties have their origin in the grain boundaries of the material. Electric charge is accumulated in the boundaries, causing a depletion layer in their vicinity and forming potential barriers (so-called Double Schottky Barriers, or DSB) which are responsible for the highly non-linear conductivity. Since ZnO is a piezoelectric material, mechanical stresses induce polarisation charges that modify the DSB heights and as a result the global electrical characteristics (i.e., piezotronic effect). In this work, a finite element method was used to simulate emerging stresses on individual grains in the bulk. Besides, experimental efforts were made to testify a coherent model that could explain this influence. Electron back scattering diffraction was used to identify grain orientations. With the help of wet chemical etching, grain polarization was determined. Micro lock-in infrared thermography (MLIRT) was applied to detect current paths through the material, and a micro 4-point probes method system (M4PPS) was employed to investigate current-voltage characteristics between single grains. Bulk samples were tested under uniaxial pressure. It was found that the conductivity can increase by up to three orders of magnitude with increasing stress. Through in-situ MLIRT, it could be shown that this effect is caused by the activation of additional current paths in the material. Further, compressive tests were performed on miniaturized samples with grain paths containing solely one or two grain boundaries. The tests evinced both an increase of the conductivity, as observed for the bulk, as well as a decreased conductivity. This phenomenon has been predicted theoretically and can be explained by piezotronically induced surface charges that have an impact on the DSB at the grain boundaries. Depending on grain orientation and stress direction, DSB can be raised or lowered. Also, the experiments revealed that the conductivity within one single specimen can increase and decrease, depending on the current direction. This novel finding indicates the existence of asymmetric Double Schottky Barriers, which was furthermore proved by complementary methods. MLIRT studies showed that the intensity of heat generation within individual current paths is dependent on the direction of the stimulating current. M4PPS was used to study the relationship between the I-V characteristics of single grain boundaries and grain orientation and revealed asymmetric behavior for very specific orientation configurations. A new model for the Double Schottky Barrier, taking into account the natural asymmetry and explaining the experimental results, will be given.Keywords: Asymmetric Double Schottky Barrier, piezotronic, varistor, zinc oxide
Procedia PDF Downloads 26858 A Novel Concept of Optical Immunosensor Based on High-Affinity Recombinant Protein Binders for Tailored Target-Specific Detection
Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly
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Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography
Procedia PDF Downloads 36857 Surface Adjustments for Endothelialization of Decellularized Porcine Pericardium
Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova
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The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF
Procedia PDF Downloads 26556 Co-Culture with Murine Stromal Cells Enhances the In-vitro Expansion of Hematopoietic Stem Cells in Response to Low Concentrations of Trans-Resveratrol
Authors: Mariyah Poonawala, Selvan Ravindran, Anuradha Vaidya
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Despite much progress in understanding the regulatory factors and cytokines that support the maturation of the various cell lineages of the hematopoietic system, factors that govern the self-renewal and proliferation of hematopoietic stem cells (HSCs) is still a grey area of research. Hematopoietic stem cell transplantation (HSCT) has evolved over the years and gained tremendous importance in the treatment of both malignant and non-malignant diseases. However, factors such as graft rejection and multiple organ failure have challenged HSCT from time to time, underscoring the urgent need for development of milder processes for successful hematopoietic transplantation. An emerging concept in the field of stem cell biology states that the interactions between the bone-marrow micro-environment and the hematopoietic stem and progenitor cells is essential for regulation, maintenance, commitment and proliferation of stem cells. Understanding the role of mesenchymal stromal cells in modulating the functionality of HSCs is, therefore, an important area of research. Trans-resveratrol has been extensively studied for its various properties to combat and prevent cancer, diabetes and cardiovascular diseases etc. The aim of the present study was to understand the effect of trans-resveratrol on HSCs using single and co-culture systems. We have used KG1a cells since it is a well accepted hematopoietic stem cell model system. Our preliminary experiments showed that low concentrations of trans-resveratrol stimulated the HSCs to undergo proliferation whereas high concentrations of trans-resveratrol did not stimulate the cells to proliferate. We used a murine fibroblast cell line, M210B4, as a stromal feeder layer. On culturing the KG1a cells with M210B4 cells, we observed that the stimulatory as well as inhibitory effects of trans-resveratrol at low and high concentrations respectively, were enhanced. Our further experiments showed that low concentration of trans-resveratrol reduced the generation of reactive oxygen species (ROS) and nitric oxide (NO) whereas high concentrations increased the oxidative stress in KG1a cells. We speculated that perhaps the oxidative stress was imposing inhibitory effects at high concentration and the same was confirmed by performing an apoptotic assay. Furthermore, cell cycle analysis and growth kinetic experiments provided evidence that low concentration of trans-resveratrol reduced the doubling time of the cells. Our hypothesis is that perhaps at low concentration of trans-resveratrol the cells get pushed into the G0/G1 phase and re-enter the cell cycle resulting in their proliferation, whereas at high concentration the cells are perhaps arrested at G2/M phase or at cytokinesis and therefore undergo apoptosis. Liquid Chromatography-Quantitative-Time of Flight–Mass Spectroscopy (LC-Q-TOF MS) analyses indicated the presence of trans-resveratrol and its metabolite(s) in the supernatant of the co-cultured cells incubated with high concentration of trans-resveratrol. We conjecture that perhaps the metabolites of trans-resveratrol are responsible for the apoptosis observed at the high concentration. Our findings may shed light on the unsolved problems in the in vitro expansion of stem cells and may have implications in the ex vivo manipulation of HSCs for therapeutic purposes.Keywords: co-culture system, hematopoietic micro-environment, KG1a cell line, M210B4 cell line, trans-resveratrol
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