Search results for: factor structure

Authors: Carlos A. C. G. Neto, Natan C. G. Silva, Thais O. Costa, Luciana R. B. Goncalves, Maria v. P. Rocha

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Galactosidases are enzymes responsible for catalyzing lactose hydrolysis reactions and also favoring transgalactosylation reactions for the production of prebiotics, among which lactulose stands out. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers in immobilization processes is a promising alternative in order to extend the useful life of the biocatalysts, for example, the coating with polyethyleneimine (PEI). PEI is a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases and also protects it from environmental variations, for example, pH and temperature. In addition, it can substantially improve the immobilization parameters and also the efficiency of enzymatic reactions. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from Kluyveromyces lactis immobilized on PEI coated agarose, determining the immobilization parameters, its operational and thermal stability, and then to apply it in the hydrolysis of lactose and synthesis of lactulose, using whey as a substrate. This immobilization strategy was chosen in order to improve the catalytic efficiency of the enzyme in the transgalactosylation reaction for the production of prebiotics, and there are few studies with β-galactosidase from this strain. The immobilization of β-galactosidase in agarose previously functionalized with 48% (w/v) glycidol and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg/g of protein. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey (66.7 g/L of lactose) supplemented with 133.3 g/L fructose at a ratio of 1:2 (lactose/fructose). Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6, with 0.1 mM MnCl2. The biocatalysts whose supports were coated were named AGA_GLY_PEI_GAL, and those that were not coated were named AGA_GLY_GAL. The coating of the support with PEI considerably improved immobilization yield (2.6-fold), the biocatalyst activity (1.4-fold), and efficiency (2.2-fold). The biocatalyst AGA_GLY_PEI_GAL was better than AGA_GLY_GAL in hydrolysis and transgalactosylation reactions, converting 88.92% of lactose at 5 min of reaction and obtaining a residual concentration of 5.24 g/L. Besides that, it was produced 13.90 g/L lactulose in the same time interval. AGA_GLY_PEI_GAL biocatalyst was stable during the 10 cycles evaluated, converting approximately 80% of lactose and producing 10.95 g/L of lactulose even after the tenth cycle. However, the thermal stability of AGA_GLY_GAL biocatalyst was superior, with a half-life time 5 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (AGA_GLY_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as the enzyme, catalyzed reactions. In addition, the use of whey as a raw material for lactulose production has proved to be an industrially advantageous alternative.

Keywords: β-galactosidase, immobilization, lactulose, polyethylenimine, whey

Procedia PDF Downloads 117

Authors: Z. Karahaliloğlu, C. Hacker, M. Demirbilek, G. Seide, E. B. Denkbaş, T. Gries

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Currently, textile industry has played an important role in world’s economy, especially in developing countries. Dyes and pigments used in textile industry are significant pollutants. Most of theirs are azo dyes that have chromophore (-N=N-) in their structure. There are many methods for removal of the dyes from wastewater such as chemical coagulation, flocculation, precipitation and ozonation. But these methods have numerous disadvantages and alternative methods are needed for wastewater decolorization. Titanium-mediated photodegradation has been used generally due to non-toxic, insoluble, inexpensive, and highly reactive properties of titanium dioxide semiconductor (TiO2). Melt electrospinning is an attractive manufacturing process for thin fiber production through electrospinning from PP (Polyprophylene). PP fibers have been widely used in the filtration due to theirs unique properties such as hydrophobicity, good mechanical strength, chemical resistance and low-cost production. In this study, we aimed to investigate the effect of titanium nanoparticle localization and amine modification on the dye degradation. The applicability of the prepared chemical activated composite and pristine fabrics for a novel treatment of dyeing wastewater were evaluated.In this study, a photocatalyzer material was prepared from nTi (titanium dioxide nanoparticles) and PP by a melt-electrospinning technique. The electrospinning parameters of pristine PP and PP/nTi nanocomposite fabrics were optimized. Before functionalization with nTi, the surface of fabrics was activated by a technique using glutaraldehyde (GA) and polyethyleneimine to promote the dye degredation. Pristine PP and PP/nTi nanocomposite melt-electrospun fabrics were characterized using scanning electron microscopy (SEM) and X-Ray Photon Spectroscopy (XPS). Methyl orange (MO) was used as a model compound for the decolorization experiments. Photocatalytic performance of nTi-loaded pristine and nanocomposite melt-electrospun filters was investigated by varying initial dye concentration 10, 20, 40 mg/L). nTi-PP composite fabrics were successfully processed into a uniform, fibrous network of beadless fibers with diameters of 800±0.4 nm. The process parameters were determined as a voltage of 30 kV, a working distance of 5 cm, a temperature of the thermocouple and hotcoil of 260–300 ºC and a flow rate of 0.07 mL/h. SEM results indicated that TiO2 nanoparticles were deposited uniformly on the nanofibers and XPS results confirmed the presence of titanium nanoparticles and generation of amine groups after modification. According to photocatalytic decolarization test results, nTi-loaded GA-treated pristine or nTi-PP nanocomposite fabric filtern have superior properties, especially over 90% decolorization efficiency at GA-treated pristine and nTi-PP composite PP fabrics. In this work, as a photocatalyzer for wastewater treatment, surface functionalized with nTi melt-electrospun fabrics from PP were prepared. Results showed melt-electrospun nTi-loaded GA-tretaed composite or pristine PP fabrics have a great potential for use as a photocatalytic filter to decolorization of wastewater and thus, requires further investigation.

Keywords: titanium oxide nanoparticles, polyprophylene, melt-electrospinning

Procedia PDF Downloads 266

Authors: Yasemin İlkay

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Street is not only a crucial spatial unit in urban design and planning discipline but also the context of walking practice in urban space. Moreover, psychogeography concentrates on both ‘walking’ and, therefore, the differentiated forms of (urban) streets to examine the influence of the built environment on the feelings and attitudes of human beings. This paper focuses on ‘walking practice’ in university campuses with reference to spatial appropriation forms via a psychogeographic lens on the phenomenon of alle in two different cities of Turkey, Ankara, the capital city, and Van, in the eastern part of the country. Alle, as an extension of ‘street’ in university campuses, is the constructive spatial structure in university campuses, and as a result, it should be the (both physical and mental) spine of design policy while conceiving and constructing a university campus. The main question of the paper is: How does the interrelation of ‘campus design’ and ‘walking practice’ on alle penetrate reciprocally on the spatial representations of citizens within their urban daily lives. The body contacts with and at urban space (with other objects and subjects) via its movements and stops; this interaction occurs through the spatial pattern of occupancy and vacancy. Walking practice leads to a set of cognitive mental representations in relation to the repertoire of place attachment and spatial appropriation. University campuses are autonomous and fruitful urban spaces to investigate such an interaction. There are both physical/real and psychogeographic representations of the same urban spaces and urban spatial practices. This separation would indicate the invisible dimensions of the difference between ‘what is conceived’ and ‘what is perceived.’ This study aims to compare and contrast the role of alle in both campus design and spatial appropriation via walking at two differentiated university campuses by collecting the mental representations, doing in-depth interviews, and attending walks with the interviewees by psychogeographic techniques. Campus design and spatial appropriation will be compared [with reference to the conception and perception of alle] in three scales: (1) the historical spatial development stories and design approaches of university campuses, (2) the spatial pattern of campuses on the basis of alle, and (3) sub-behavioral regions of the alle in campuses in relation with mental representations and psychogeographic attentive walks. The sub-questions of the research are: [1] How and why do the design approaches differentiate in two university campuses in Turkey, [2] How the interrelation among alle design and spatial appropriation differs in these two cases, and [3] What do the differentiated gaps among real and psychographic maps indicate about the design and spatial appropriation interrelation. METU, as a well-designed, readable campus with its alle, promise a rich walking practice with in-depth and fruitful spatial appropriation regions; however, Van YYÜ limits both the practice and place attachment with its partial design with an alle which is later added to the campus. This research both displays the role of alle in the campus design, walking practice and spatial appropriation and opens a new methodological path to discover hidden knowledge within urban spaces.

Keywords: alle, campus design, cognitive geography, psychogeography, spatial appropriation, Turkey

Procedia PDF Downloads 100

Authors: Irina P. Starodubtseva, Aleksandr N. Pavlenko

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Quenching is a term generally accepted for the process of rapid cooling of a solid that is overheated above the thermodynamic limit of the liquid superheat. The main objective of many previous studies on quenching is to find a way to reduce the total time of the transient process. Computational experiments were performed to simulate quenching by a falling liquid nitrogen film of an extremely overheated vertical copper plate with a structured capillary-porous coating. The coating was produced by directed plasma spraying. Due to the complexities in physical pattern of quenching from chaotic processes to phase transition, the mechanism of heat transfer during quenching is still not sufficiently understood. To our best knowledge, no information exists on when and how the first stable liquid-solid contact occurs and how the local contact area begins to expand. Here we have more models and hypotheses than authentically established facts. The peculiarities of the quench front dynamics and heat transfer in the transient process are studied. The created numerical model determines the quench front velocity and the temperature fields in the heater, varying in space and time. The dynamic pattern of the running quench front obtained numerically satisfactorily correlates with the pattern observed in experiments. Capillary-porous coatings with straight and reverse orientation of crests are investigated. The results show that the cooling rate is influenced by thermal properties of the coating as well as the structure and geometry of the protrusions. The presence of capillary-porous coating significantly affects the dynamics of quenching and reduces the total quenching time more than threefold. This effect is due to the fact that the initialization of a quench front on a plate with a capillary-porous coating occurs at a temperature significantly higher than the thermodynamic limit of the liquid superheat, when a stable solid-liquid contact is thermodynamically impossible. Waves present on the liquid-vapor interface and protrusions on the complex micro-structured surface cause destabilization of the vapor film and the appearance of local liquid-solid micro-contacts even though the average integral surface temperature is much higher than the liquid superheat limit. The reliability of the results is confirmed by direct comparison with experimental data on the quench front velocity, the quench front geometry, and the surface temperature change over time. Knowledge of the quench front velocity and total time of transition process is required for solving practically important problems of nuclear reactors safety.

Keywords: capillary-porous coating, heat transfer, Leidenfrost phenomenon, numerical simulation, quenching

Procedia PDF Downloads 129

Authors: Amanda Gregorim Fernandes, Lorena Cardoso Cintra, Rosalia Santos Amorim Jesuino, Fabricia Paula De Faria, Marcio José Poças Fonseca

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Bioethanol is one of the most promising biofuels and able to replace fossil fuels and reduce its different environmental impacts and can be generated from various agroindustrial waste. The Brazil is in first place in bioethanol production to be the largest producer of sugarcane. The bagasse sugarcane (SCB) has lignocellulose which is composed of three major components: cellulose, hemicellulose and lignin. Cellulose is a homopolymer of glucose units connected by glycosidic linkages. Among all species of Penicillium, Penicillium echinulatum has been the focus of attention because they produce high quantities of cellulase and the mutant strain 9A02S1 produces higher enzyme levels compared to the wild. Among the cellulases, the cellobiohydrolases enzymes are the main components of the cellulolytic system of fungi, and are also responsible for most of the potential hydrolytic in enzyme cocktails for the industrial processing of plant biomass and several cellobiohydrolases Penicillium had higher specific activity against cellulose compared to CBH I from Trichoderma reesei. This fact makes it an interesting pattern for higher yields in the enzymatic hydrolysis, and also they are important enzymes in the hydrolysis of crystalline regions of cellulose. Therefore, finding new and more active enzymes become necessary. Meanwhile, β-glycosidases act on soluble substrates and are highly dependent on cellobiohydrolases and endoglucanases action to provide the substrate in the hydrolysis of the biomass, but the cellobiohydrolases and endoglucanases are highly dependent β-glucosidases to maintain efficient hydrolysis. Thus, there is a need to understand the structure-function relationships that govern the catalytic activity of cellulolytic enzymes to elucidate its mechanism of action and optimize its potential as industrial biocatalysts. To evaluate the enzyme β-glucosidase of Penicillium echinulatum (PeBGL1) the gene was synthesized from the assembly sequence from a library in induction conditions and then the PeBGL1 gene was cloned in the vector pPICZαA and transformed into P. pastoris GS115. After processing, the producers of PeBGL1 were analyzed for enzyme activity and protein profile where a band of approximately 100 kDa was viewed. It was also carried out the zymogram. In partial characterization it was determined optimum temperature of 50°C and optimum pH of 6,5. In addition, to increase the secreted recombinant PeBGL1 production by Pichia pastoris, three parameters of P. pastoris culture medium were analysed: methanol, nitrogen source concentrations and the inoculum size. A 23 factorial design was effective in achieving the optimum condition. Altogether, these results point to the potential application of this P. echinulatum β-glucosidase in hydrolysis of cellulose for the production of bioethanol.

Keywords: bioethanol, biotechnology, beta-glucosidase, penicillium echinulatum

Procedia PDF Downloads 241

Authors: Bipasa Dey, Varsha Panwar, Tanmay Dutta

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Laccase, a multicopper oxidase (EC 1.10.3.2) have been at the forefront as a superior industrial biocatalyst. They are versatile in terms of bestowing sustainable and ecological catalytic reactions such as polymerisation, xenobiotic degradation and bioremediation of phenolic and non-phenolic compounds. Regardless of the wide biotechnological applications, the critical limiting factors viz. reusability, retrieval, and storage stability still prevail. This can cause an impediment in their applicability. Crosslinked enzyme aggregates (CLEAs) have emerged as a promising technique that rehabilitates these essential facets, albeit at the expense of their enzymatic activity. The carrier free crosslinking method prevails over the carrier-bound immobilisation in conferring high productivity, low production cost owing to the absence of additional carrier and circumvent any non-catalytic ballast which could dilute the volumetric activity. To the best of our knowledge, the ε-amino group of lysyl residue is speculated as the best choice for forming Schiff’s base with glutaraldehyde. Despite being most preferrable, excess glutaraldehyde can bring about disproportionate and undesirable crosslinking within the catalytic site and hence could deliver undesirable catalytic losses. Moreover, the surface distribution of lysine residues in Trametes versicolor laccase is significantly less. Thus, to mitigate the adverse effect of glutaraldehyde in conjunction with scaling down the degradation or catalytic loss of the enzyme, crosslinking with inert substances like gelatine, collagen, Bovine serum albumin (BSA) or excess lysine is practiced. Analogous to these molecules, sugars have been well known as a protein stabiliser. It helps to retain the structural integrity, specifically secondary structure of the protein during aggregation by changing the solvent properties. They are comprehended to avert protein denaturation or enzyme deactivation during precipitation. We prepared crosslinked enzyme aggregates (CLEAs) of laccase from T. versicolor with the aid of sugars. The sugar CLEAs were compared with the classic BSA and glutaraldehyde laccase CLEAs concerning physico-chemical properties. The activity recovery for the fructose CLEAs were found to be ~20% higher than the non-sugar CLEA. Moreover, the 𝐾𝑐𝑎𝑡𝐾𝑚⁄ values of the CLEAs were two and three-fold higher than BSA-CLEA and GACLEA, respectively. The half-life (t1/2) deciphered by sugar-CLEA was higher than the t1/2 of GA-CLEAs and free enzyme, portraying more thermal stability. Besides, it demonstrated extraordinarily high pH stability, which was analogous to BSA-CLEA. The promising attributes of increased storage stability and recyclability (>80%) gives more edge to the sugar-CLEAs over conventional CLEAs of their corresponding free enzyme. Thus, sugar-CLEA prevails in furnishing the rudimentary properties required for a biocatalyst and holds many prospects.

Keywords: cross-linked enzyme aggregates, laccase immobilization, enzyme reusability, enzyme stability

Procedia PDF Downloads 100

Authors: M. Vezir Kahraman, Emre Basturk

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Energy storage technology offers new ways to meet the demand to obtain efficient and reliable energy storage materials. Thermal energy storage systems provide the potential to acquire energy savings, which in return decrease the environmental impact related to energy usage. For this purpose, phase change materials (PCMs) that work as 'latent heat storage units' which can store or release large amounts of energy are preferred. Phase change materials (PCMs) are being utilized to absorb, collect and discharge thermal energy during the cycle of melting and freezing, converting from one phase to another. Phase Change Materials (PCMs) can generally be arranged into three classes: organic materials, salt hydrates and eutectics. Many kinds of organic and inorganic PCMs and their blends have been examined as latent heat storage materials. PCMs have found different application areas such as solar energy storage and transfer, HVAC (Heating, Ventilating and Air Conditioning) systems, thermal comfort in vehicles, passive cooling, temperature controlled distributions, industrial waste heat recovery, under floor heating systems and modified fabrics in textiles. Ultraviolet (UV)-curing technology has many advantages, which made it applicable in many different fields. Low energy consumption, high speed, room-temperature operation, low processing costs, high chemical stability, and being environmental friendly are some of its main benefits. UV-curing technique has many applications. One of the many advantages of UV-cured PCMs is that they prevent the interior PCMs from leaking. Shape-stabilized PCM is prepared by blending the PCM with a supporting material, usually polymers. In our study, this problem is minimized by coating the fatty alcohols with a photo-cross-linked thiol-ene based polymeric system. Leakage is minimized because photo-cross-linked polymer acts a matrix. The aim of this study is to introduce a novel thiol-ene based shape-stabilized PCM. Photo-crosslinked thiol-ene based polymers containing fatty alcohols were prepared and characterized for the purpose of phase change materials (PCMs). Different types of fatty alcohols were used in order to investigate their properties as shape-stable PCMs. The structure of the PCMs was confirmed by ATR-FTIR techniques. The phase transition behaviors, thermal stability of the prepared photo-crosslinked PCMs were investigated by differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). This work was supported by Marmara University, Commission of Scientific Research Project.

Keywords: differential scanning calorimetry (DSC), Polymeric phase change material, thermal energy storage, UV-curing

Procedia PDF Downloads 227

Authors: Alejandra Martin-Dominguez, Javier Canto-Rios, Velitchko Tzatchkov

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Electrocoagulation is a water treatment technology that consists of generating coagulant species in situ by electrolytic oxidation of sacrificial anode materials triggered by electric current. It removes suspended solids, heavy metals, emulsified oils, bacteria, colloidal solids and particles, soluble inorganic pollutants and other contaminants from water, offering an alternative to the use of metal salts or polymers and polyelectrolyte addition for breaking stable emulsions and suspensions. The method essentially consists of passing the water being treated through pairs of consumable conductive metal plates in parallel, which act as monopolar electrodes, commonly known as ‘sacrificial electrodes’. Physicochemical, electrochemical and hydraulic processes are involved in the efficiency of this type of treatment. While the physicochemical and electrochemical aspects of the technology have been extensively studied, little is known about the influence of the hydraulics. However, the hydraulic process is fundamental for the reactions that take place at the electrode boundary layers and for the coagulant mixing. Electrocoagulation reactors can be open (with free water surface) and closed (pressurized). Independently of the type of rector, hydraulic head loss is an important factor for its design. The present work focuses on the study of the total hydraulic head loss and flow velocity and pressure distribution in electrocoagulation reactors with single or multiple concentric annular cross sections. An analysis of the head loss produced by hydraulic wall shear friction and accessories (minor head losses) is presented, and compared to the head loss measured on a semi-pilot scale laboratory model for different flow rates through the reactor. The tests included laminar, transitional and turbulent flow. The observed head loss was compared also to the head loss predicted by several known conceptual theoretical and empirical equations, specific for flow in concentric annular pipes. Four single concentric annular cross section and one multiple concentric annular cross section reactor configuration were studied. The theoretical head loss resulted higher than the observed in the laboratory model in some of the tests, and lower in others of them, depending also on the assumed value for the wall roughness. Most of the theoretical models assume that the fluid elements in all annular sections have the same velocity, and that flow is steady, uniform and one-dimensional, with the same pressure and velocity profiles in all reactor sections. To check the validity of such assumptions, a computational fluid dynamics (CFD) model of the concentric annular pipe reactor was implemented using the ANSYS Fluent software, demonstrating that pressure and flow velocity distribution inside the reactor actually is not uniform. Based on the analysis, the equations that predict better the head loss in single and multiple annular sections were obtained. Other factors that may impact the head loss, such as the generation of coagulants and gases during the electrochemical reaction, the accumulation of hydroxides inside the reactor, and the change of the electrode material with time, are also discussed. The results can be used as tools for design and scale-up of electrocoagulation reactors, to be integrated into new or existing water treatment plants.

Keywords: electrocoagulation reactors, hydraulic head loss, concentric annular pipes, computational fluid dynamics model

Procedia PDF Downloads 217

Authors: Susanna Graziano, Lydia Krstic Ward

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The purpose of this presentation is to demonstrate the steps of using multimedia and collaborative platforms in project-based learning. The presentation will demonstrate the stages of the learning project with various components of independent and collaborative learning, where students research the topic, share information, prepare a survey, use social media (Facebook, Instagram, WhasApp) and collaborative platforms (wikispaces.com and Google docs) to collect, analyze and process data, then produce reports and logos to be displayed as a final product. At the beginning of the presentation participants will answer a questionnaire about project based learning and share their experience on using social media, real–world project work and collaborative learning. Using a PPP, the presentation will walk participants through the steps of a completed project where tertiary education students are involved in putting together a multimedia campaign for safe driving in Kuwait. The research component of the project entails taking a holistic view on the problem of the high death rate in traffic accidents. The final goal of the project is to lead students to raise public awareness about the importance of safe driving. The project steps involve using the social media and collaborative platforms for collecting data and sharing the required materials to be used in the final product – a display of written reports, slogans and videos, as well as oral presentations. The same structure can be used to organize a multimedia campaign focusing on other issues, whilst scaffolding on students’ ability to brainstorm, retrieve information, organize it and engage in collaborative/ cooperative learning whilst being immersed in content-based learning as well as in authentic tasks. More specifically, the project we carried out at Box Hill College was a real-world one and involved a multimedia Campaign for Safe Driving since reckless driving is one of the major problems in the country. The idea for the whole project started by a presentation given by a board member of the Kuwaiti Society for Traffic Safety who was invited to college and spoke about: • Driving laws in the country, • What causes car accidents, • Driving safety tips. The principal goal of this project was to let students consider problems of traffic in Kuwait from different points of view. They also had to address the number and causes of accidents, evaluate the effectiveness of the local traffic law in order to send a warning about the importance of safe driving and, finally, suggest ways of its improvement. Benefits included: • Engagement, • Autonomy, • Motivation, • Content knowledge, • Language mastery, • Enhanced critical thinking, • Increased metacognitive awareness, • Improved social skills, • Authentic experience.

Keywords: social media, online learning platforms, collaborative platforms, project based learning

Procedia PDF Downloads 424

Authors: Xiao-Mei Zhang, Sian-Hong Tseng, Ming-Yen Lu

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Two-dimensional (2D) transition metal dichalcogenides (TMDCs), such as MoS₂, have attracted considerable attention owing to the unique optical and electronic properties related to its 2D ultrathin atomic layer structure. MoS₂ is becoming prevalent in post-silicon digital electronics and in highly efficient optoelectronics due to its extremely low thickness and its tunable band gap (Eg = 1-2 eV). For low-power, high-performance complementary logic applications, both p- and n-type MoS₂ FETs (NFETs and PFETs) must be developed. NFETs with an electron accumulation channel can be obtained using unintentionally doped n-type MoS₂. However, the fabrication of MoS₂ FETs with complementary p-type characteristics is challenging due to the significant difficulty of injecting holes into its inversion channel. Plasma treatments with different species (including CF₄, SF₆, O₂, and CHF₃) have also been found to achieve the desired property modifications of MoS₂. In this work, we demonstrated a p-type multilayer MoS₂ enabled by selective-area doping using CHF₃ plasma treatment. Compared with single layer MoS₂, multilayer MoS₂ can carry a higher drive current due to its lower bandgap and multiple conduction channels. Moreover, it has three times the density of states at its minimum conduction band. Large-area growth of MoS₂ films on 300 nm thick SiO₂/Si substrate is carried out by thermal decomposition of ammonium tetrathiomolybdate, (NH₄)₂MoS₄, in a tube furnace. A two-step annealing process is conducted to synthesize MoS₂ films. For the first step, the temperature is set to 280 °C for 30 min in an N₂ rich environment at 1.8 Torr. This is done to transform (NH₄)₂MoS₄ into MoS₃. To further reduce MoS₃ into MoS₂, the second step of annealing is performed. For the second step, the temperature is set to 750 °C for 30 min in a reducing atmosphere consisting of 90% Ar and 10% H₂ at 1.8 Torr. The grown MoS₂ films are subjected to out-of-plane doping by CHF₃ plasma treatment using a Dry-etching system (ULVAC original NLD-570). The radiofrequency power of this dry-etching system is set to 100 W and the pressure is set to 7.5 mTorr. The final thickness of the treated samples is obtained by etching for 30 s. Back-gated MoS₂ PFETs were presented with an on/off current ratio in the order of 10³ and a field-effect mobility of 65.2 cm²V⁻¹s⁻¹. The MoS₂ PFETs photodetector exhibited ultraviolet (UV) photodetection capability with a rapid response time of 37 ms and exhibited modulation of the generated photocurrent by back-gate voltage. This work suggests the potential application of the mild plasma-doped p-type multilayer MoS₂ in UV photodetectors for environmental monitoring, human health monitoring, and biological analysis.

Keywords: photodetection, p-type doping, multilayers, MoS₂

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Authors: Rituparna Pal, Faiz Ahmed

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Neighbourhood energy efficiency is a newly emerged term to address the quality of urban strata of built environment in terms of various covariates of sustainability. The concept of sustainability paradigm in developed nations has encouraged the policymakers for developing urban scale cities to envision plans under the aegis of urban scale sustainability. The concept of neighbourhood energy efficiency is realized a lot lately just when the cities, towns and other areas comprising this massive global urban strata have started facing a strong blow from climate change, energy crisis, cost hike and an alarming shortfall in the justice which the urban areas required. So this step of urban sustainability can be easily referred more as a ‘Retrofit Action’ which is to cover up the already affected urban structure. So even if we start energy efficiency for existing cities and urban areas the initial layer remains, for which a complete model of urban sustainability still lacks definition. Urban sustainability is a broadly spoken off word with end number of parameters and policies through which the loop can be met. Out of which neighbourhood energy efficiency can be an integral part where the concept and index of neighbourhood scale indicators, block level indicators and building physics parameters can be understood, analyzed and concluded to help emerge guidelines for urban scale sustainability. The future of neighbourhood energy efficiency not only lies in energy efficiency but also important parameters like quality of life, access to green, access to daylight, outdoor comfort, natural ventilation etc. So apart from designing less energy-hungry buildings, it is required to create a built environment which will create less stress on buildings to consume more energy. A lot of literary analysis has been done in the Western countries prominently in Spain, Paris and also Hong Kong, leaving a distinct gap in the Indian scenario in exploring the sustainability at the urban strata. The site for the study has been selected in the upcoming capital city of Amaravati which can be replicated with similar neighbourhood typologies in the area. The paper suggests a methodical intent to quantify energy and sustainability indices in detail taking by involving several macro, meso and micro level covariates and parameters. Several iterations have been made both at macro and micro level and have been subjected to simulation, computation and mathematical models and finally to comparative analysis. Parameters at all levels are analyzed to suggest the best case scenarios which in turn is extrapolated to the macro level finally coming out with a proposal model for energy efficient neighbourhood and worked out guidelines with significance and correlations derived.

Keywords: energy quantification, macro scale parameters, meso scale parameters, micro scale parameters

Procedia PDF Downloads 175

Authors: Sijie Chen, Xiaofang Chen, Juan Wang, Yingying Zhang, Yu Hong, Wanyu Zhuang, Xinxin Huang, Ping Ou, Longsheng Huang

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Purpose: Autistic symptom improvement can be observed in children treated with acupuncture, but the mechanism is still being explored. In the present study, we used scalp acupuncture to treat autism rat model, and then their improvement in the abnormal behaviors and specific mechanisms behind were revealed by detecting animal behaviors, analyzing the RNA sequencing of the prefrontal cortex(PFC), and observing the ultrastructure of PFC neurons under the transmission electron microscope. Methods: On gestational day 12.5, Wistar rats were given valproic acid (VPA) by intraperitoneal injection, and their offspring were considered to be reliable rat models of autism. They were randomized to VPA or VPA-acupuncture group (n=8). Offspring of Wistar pregnant rats that were simultaneously injected with saline were randomly selected as the wild-type group (WT). VPA_acupuncture group rats received acupuncture intervention at 23 days of age for 4 weeks, and the other two groups followed without intervention. After the intervention, all experimental rats underwent behavioral tests. Immediately afterward, they were euthanized by cervical dislocation, and their prefrontal cortex was isolated for RNA sequencing and transmission electron microscopy. Results: The main results are as follows: 1. Animal behavioural tests: VPA group rats showed more anxiety-like behaviour and repetitive, stereotyped behaviour than WT group rats. While VPA group rats showed less spatial exploration ability, activity level, social interaction, and social novelty preference than WT group rats. It was gratifying to observe that acupuncture indeed improved these abnormal behaviors of autism rat model. 2. RNA-sequencing: The three groups of rats differed in the expression and enrichment pathways of multiple genes related to synaptic function, neural signal transduction, and circadian rhythm regulation. Our experiments indicated that acupuncture can alleviate the major symptoms of ASD by improving these neurological abnormalities. 3. Under the transmission electron microscopy, several lysosomes and mitochondrial structural abnormalities were observed in the prefrontal neurons of VPA group rats, which were manifested as atrophy of the mitochondrial membran, blurring or disappearance of the mitochondrial cristae, and even vacuolization. Moreover, the number of synapses and synaptic vesicles was relatively small. Conversely, the mitochondrial structure of rats in the WT group and VPA_acupuncture was normal, and the number of synapses and synaptic vesicles was relatively large. Conclusion: Acupuncture effectively improved the abnormal behaviors of autism rat model and the ultrastructure of the PFC neurons, which might worked by improving their abnormal synaptic function, synaptic plasticity and promoting neuronal signal transduction.

Keywords: autism spectrum disorder, acupuncture, animal behavior, RNA sequencing, transmission electron microscope

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Authors: Nirjhar Dhang, S. Vinay Kumar

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Concrete is a non-homogeneous mix of coarse aggregates, sand, cement, air-voids and interfacial transition zone (ITZ) around aggregates. Adoption of these complex structures and material properties in numerical simulation would lead us to better understanding and design of concrete. In this work, the mesoscale model of concrete has been prepared from X-ray computerized tomography (CT) image. These images are converted into computer model and numerically simulated using commercially available finite element software. The mesoscale models are simulated under the influence of compressive displacement. The effect of shape and distribution of aggregates, continuous and discrete ITZ thickness, voids, and variation of mortar strength has been investigated. The CT scan of concrete cube consists of series of two dimensional slices. Total 49 slices are obtained from a cube of 150mm and the interval of slices comes approximately 3mm. In CT scan images, the same cube can be CT scanned in a non-destructive manner and later the compression test can be carried out in a universal testing machine (UTM) for finding its strength. The image processing and extraction of mortar and aggregates from CT scan slices are performed by programming in Python. The digital colour image consists of red, green and blue (RGB) pixels. The conversion of RGB image to black and white image (BW) is carried out, and identification of mesoscale constituents is made by putting value between 0-255. The pixel matrix is created for modeling of mortar, aggregates, and ITZ. Pixels are normalized to 0-9 scale considering the relative strength. Here, zero is assigned to voids, 4-6 for mortar and 7-9 for aggregates. The value between 1-3 identifies boundary between aggregates and mortar. In the next step, triangular and quadrilateral elements for plane stress and plane strain models are generated depending on option given. Properties of materials, boundary conditions, and analysis scheme are specified in this module. The responses like displacement, stresses, and damages are evaluated by ABAQUS importing the input file. This simulation evaluates compressive strengths of 49 slices of the cube. The model is meshed with more than sixty thousand elements. The effect of shape and distribution of aggregates, inclusion of voids and variation of thickness of ITZ layer with relation to load carrying capacity, stress-strain response and strain localizations of concrete have been studied. The plane strain condition carried more load than plane stress condition due to confinement. The CT scan technique can be used to get slices from concrete cores taken from the actual structure, and the digital image processing can be used for finding the shape and contents of aggregates in concrete. This may be further compared with test results of concrete cores and can be used as an important tool for strength evaluation of concrete.

Keywords: concrete, image processing, plane strain, interfacial transition zone

Procedia PDF Downloads 238

Authors: Antonio J. Melendez-Martinez

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Beyond their role as natural colorants, some carotenoids are provitamins A and may be involved in health-promoting biological actions and contribute to reducing the risk of developing non-communicable diseases, including several types of cancer, cardiovascular disease, eye conditions, skin disorders or metabolic disorders. Given the versatility of carotenoids, the COST-funded European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN) is aimed at promoting health through the diet and increasing well-being by means. Stakeholders from 38 countries participate in this network, and one of its main objectives is to promote research on little-studied carotenoids. In this contribution, recent advances of our research group and collaborators in the study of two such understudied carotenoids, namely phytoene and phytofluene, the colorless carotenoids, are outlined. The study of these carotenoids is important as they have been largely neglected despite they are present in our diets, fluids, and tissues, and evidence is accumulating that they may be involved in health-promoting actions. More specifically, studies on their levels in diverse tomato and orange varieties were carried out as well as on their potential bioavailability from different dietary sources. Furthermore, the potential effect of these carotenoids on an animal model subjected to oxidative stress was evaluated. The tomatoes were grown in research greenhouses, and some of them were subjected to regulated deficit irrigation, a sustainable agronomic practice. The citrus samples were obtained from an experimental field. The levels of carotenoids were assessed using HPLC according to routine methodologies followed in our lab. Regarding the potential bioavailability (bioaccessibility) studies, different products containing colorless carotenoids, like fruits, juices, were subjected to simulated in vitro digestions, and their incorporation into mixed micelles was assessed. The effect of the carotenoids on oxidative stress was evaluated on the Caenorhabditis elegans model. For that purpose, the worms were subjected to oxidative stress by means of a hydrogen peroxide challenge. In relation to the presence of colorless carotenoids in tomatoes and orange varieties, it was observed that they are widespread in such products and that there are mutants with very high quantities of them, for instance, the Cara Cara or Pinalate mutant oranges. The studies on their bioaccessibility revealed that, in general, phytoene and phytofluene are more bioaccessible than other common dietary carotenoids, probably due to their distinctive chemical structure. About the in vivo antioxidant capacity of phytoene and phytofluene, it was observed that they both exerted antioxidant effects at certain doses. In conclusion, evidence on the importance of phytoene and phytofluene as dietary easily bioavailable and antioxidant carotenoids has been obtained in recent studies from our group, which can be important shortly to innovate in health-promotion through the development of functional foods and related products.

Keywords: carotenoids, health, functional foods, nutrition, phytoene, phytofluene

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Authors: Samanta Kietytė

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The object of this study is the verbal derivatives stemming from the Lithuanian prefix per-. The prefix under examination can be classified as prepositional, having descended from the preposition per, thereby sharing the same prototypical meaning – denoting movement OVER. These frequently co-occur within sentences (1). The aim of this paper is to conduct a semantic analysis of the prefix per- and to propose a possible radial network of its meanings. In essence, the aim is to identify the interrelationships existing between its meanings. 1) Jis peršoko per tvorą/ 3SG.NOM.M jump.PST.3 over fence.ACC.SG. /ʻHe jumped over the fenceʼ. The foundation of this work lies in the methodological and theoretical framework of cognitive linguistics. The prototypical meaning of prefixes consistently embodies spatial dimensions that can be described through image schemas. This entails the identification of the trajectory, the landmark, and the relation between them in the situation described by the prefixed verb. The meanings of linguistic units are not perceived as arbitrary, but rather, they are interconnected through semantic motivation. According to this perspective, a singular meaning within linguistic units is considered as prototypical, while additional meanings are descended (not necessarily directly) from it. For example, one of the per- meanings TRANSFER (2) is derived from the prototypical meaning OVER. 2) Prašau persiųsti vadovo laišką man./ Ask.PRS.1 forward.INF manager.GEN.SG email.ACC.SG 1.SG.DAT/ ʻPlease forward the manager‘s email to meʼ. Certain semantic relations are explained by the conceptual metaphor and metonymy theory. For instances, when prefixed verb has a meaning WIN (3) it is related to the prototypical meaning. In this case, the prefixed verb describes situations of winning in various ways. In the prototypical meaning, the trajector moves higher than the landmark, and winning is metaphorically perceived as being higher. 3) Sūnus peraugo tėvą./ Son.NOM.SG outgrow.PST.3 father.ACC.SG/ ʻThe son has outgrown the fatherʼ. The data utilized for this study was collected from the 2014 grammatically annotated text "Lithuanian Web (LithuanianWaC v2)", consisting of 63,645,700 words. Given that the corpus is grammatically lemmatized, the list of the 793 items was obtained using the wordlist function and specifying that verbs starting with per were searched. The list included not only prefixed verbs but also other verbs whose roots have the same letter sequences as prefixes. Also, words with misspellings, without diacritical marks, and words listed for lemmatization errors were rejected, and a total of 475 derivatives were left for further analysis. The semantic analysis revealed that there are 12 distinct meanings of the prefix per-. The spatial meanings were extracted by determining what a trajector is, what a landmark is, and what the relation between them is. The connection between non-spatial meanings and spatial ones occurs through semantic motivation established by identifying elements that correspond to the trajector and landmark. The analysis reveals that there are no strict boundaries among these meanings, instead showing a continuum that encompasses a central core and a peripheral association with their internal structure, i.e., some derivatives are more prototypical of a particular meaning than others.

Keywords: word-formation, cognitive semantics, metaphor, radial networks, prototype theory, prefix

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Authors: Malory Jonata

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Chlordecone (CLD) is an organochlorine pesticide used between 1971 and 1993 in both Guadeloupe and Martinique for the control of banana black weevil. The bishomocubane structure which characterizes this chemical compound led to high stability in organic matter and high persistence in the environment. Recently, researchers found that CLD can be degraded by isolated bacteria consortiums and, particularly, by bacteria such as Citrobacter sp 86 and Delsulfovibrio sp 86. Actually, six transformation product families of CLD are known. Moreover, the latest discovery showed that CLD was disappearing faster than first predicted in highly contaminated soil in Guadeloupe. However, the toxicity of transformation products is still unknown, and knowledge has to be deepened on the degradation ways and chemical characteristics of chlordecone and its transformation products. Microbial fuel cells (MFC) are electrochemical systems that can convert organic matter into electricity thanks to electroactive bacteria. These bacteria can exchange electrons through their membranes to solid surfaces or molecules. MFC have proven their efficiency as bioremediation systems in water and soils. They are already used for the bioremediation of several organochlorine compounds such as perchlorate, trichlorophenol or hexachlorobenzene. In this study, a three-electrodes system, inspired by MFC, is used to try to degrade chlordecone using bacteria from a mangrove swamp in Martinique. As we know, some mangrove bacteria are electroactive. Furthermore, the CLD rate seems to decline in mangrove swamp sediments. This study aims to prove that electroactive bacteria from a mangrove swamp in Martinique can degrade CLD thanks to a three-electrodes bioelectrochemical system. To achieve this goal, the tree-electrodes assembly has been connected to a potentiostat. The substrate used is mangrove water and sediments sampled in the mangrove swamp of La Trinité, a coastal city in Martinique, where CLD contamination has already been studied. Electroactive biofilms are formed by imposing a potential relative to Saturated Calomel Electrode using chronoamperometry. Moreover, their comportment has been studied by using cyclic voltametry. Biofilms have been studied under different imposed potentials, several conditions of the substrate and with or without CLD. In order to quantify the evolution of CLD rates in the substrate’s system, gas chromatography coupled with mass spectrometry (GC-MS) was performed on pre-treated samples of water and sediments after short, medium and long-term contact with the electroactive biofilms. Results showed that between -0,8V and -0,2V, the three-electrodes system was able to reduce the chemical in the substrate solution. The first GC-MS analysis result of samples spiked with CLD seems to reveal decreased CLD concentration over time. In conclusion, the designed bioelectrochemical system can provide the necessary conditions for chlordecone degradation. However, it is necessary to improve three-electrodes control settings in order to increase degradation rates. The biological pathways are yet to enlighten by biologicals analysis of electroactive biofilms formed in this system. Moreover, the electrochemical study of mangrove substrate gives new informations on the potential use of this substrate for bioremediation. But further studies are needed to a better understanding of the electrochemical potential of this environment.

Keywords: bioelectrochemistry, bioremediation, chlordecone, mangrove swamp

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Authors: Sri Sai Ramya Bojedla, Falguni Pati

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Nature provides the best of solutions to humans. One such incredible gift to regenerative medicine is silk. The literature has publicized a long appreciation for silk owing to its incredible physical and biological assets. Its bioactive nature, unique mechanical strength, and processing flexibility make us curious to explore further to apply it in the clinics for the welfare of mankind. In this study, Antheraea mylitta and Bombyx mori silk fibroin microfibers are developed by two economical and straightforward steps via degumming and hydrolysis for the first time, and a bioactive composite is manufactured by mixing silk fibroin microfibers at various concentrations with polycaprolactone (PCL), a biocompatible, aliphatic semi-crystalline synthetic polymer. Reconstructive surgery in any part of the body except for the maxillofacial region deals with replacing its function. But answering both the aesthetics and function is of utmost importance when it comes to facial reconstruction as it plays a critical role in the psychological and social well-being of the patient. The main concern in developing adequate bone graft substitutes or a scaffold is the noteworthy variation in each patient's bone anatomy. Additionally, the anatomical shape and size will vary based on the type of defect. The advent of additive manufacturing (AM) or 3D printing techniques to bone tissue engineering has facilitated overcoming many of the restraints of conventional fabrication techniques. The acquired patient's CT data is converted into a stereolithographic (STL)-file which is further utilized by the 3D printer to create a 3D scaffold structure in an interconnected layer-by-layer fashion. This study aims to address the limitations of currently available materials and fabrication technologies and develop a customized biomaterial implant via 3D printing technology to reconstruct complex form, function, and aesthetics of the facial anatomy. These composite scaffolds underwent structural and mechanical characterization. Atomic force microscopic (AFM) and field emission scanning electron microscopic (FESEM) images showed the uniform dispersion of the silk fibroin microfibers in the PCL matrix. With the addition of silk, there is improvement in the compressive strength of the hybrid scaffolds. The scaffolds with Antheraea mylitta silk revealed higher compressive modulus than that of Bombyx mori silk. The above results of PCL-silk scaffolds strongly recommend their utilization in bone regenerative applications. Successful completion of this research will provide a great weapon in the maxillofacial reconstructive armamentarium.

Keywords: compressive modulus, 3d printing, maxillofacial reconstruction, natural fiber reinforced composites, silk fibroin microfibers

Procedia PDF Downloads 193

Authors: S. Jayasinghe, W. G. D. Wickramasingha, V. Karunaratne, D. N. Karunaratne, A. Ekanayake

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Multi-drug resistant microbial pathogens are a serious global health problem, and hence, there is an urgent necessity for discovering new drug therapeutics. However, finding alternatives is a one of the biggest challenges faced by the global drug industry due to the spiraling high cost and serious side effects associated with modern medicine. On the other hand, plants and their secondary metabolites can be considered as good sources of scaffolds to provide structurally diverse bioactive compounds as potential therapeutic agents. 6β-hydroxy betunolic acid is a triterpenoid isolated from bark of Schumacheria castaneifolia which is an endemic plant to Sri Lanka which has shown antibacterial activity against both Staphylococcus aureus (ATCC 29213) and methicillin-resistant S. aureus with Minimum Inhibition Concentration (MIC) of 16 µg/ml. The objective of this study was to determine the anti-bacterial activity for the derivatives of 6β- hydroxy betunolic acid against standard strains of Staphylococcus aureus (ATCC 29213 and ATCC 25923), Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 35218 and ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), carbepenemas produce Kebsiella pneumonia (ATCC BAA 1705) and carbepenemas non produce Kebsiella pneumonia (ATCC BAA 1706) and four stains of clinically isolated methicillin resistance S. aureus and Acinetobacter. Structural analogues of 6β-hydroxy betunolic acid were synthesized by modifying the carbonyl group at C-3 to obtain olefin and oxime, the hydroxyl group at C-6 position to a ketone, the carboxylic acid at C-17 to obtain amide and halo ester and the olefin group at C-20 position to obtain epoxide. Chemical structures of the synthesized analogues were confirmed with spectroscopic data and antibacterial activity was determined through broth micro dilution assay. Results revealed that 6β- hydroxy betunolic acid shows significant antibacterial activity only against the Gram positive strains and it was inactive against all the tested Gram negative strains for the tested concentration range. However, structural modifications into oxime and olefin at C-3, ketone at C-6 and epoxide at C-20 decreased its antibacterial activity against the gram positive organisms and it was totally lost with the both modifications at C-17 into amide and ester. These results concluded that the antibacterial activity of 6β- hydroxy betunolic acid and derivatives is predominantly depending on the cell wall difference of the bacteria and the presence of carboxylic acid at C-17 is highly important for the antibacterial activity against Gram positive organisms.

Keywords: antibacterial activity, 6β- hydroxy betunolic acid, broth micro dilution assay, structure activity relationship

Procedia PDF Downloads 125

Authors: Maria Angela Bedini, Fabio Bronzini

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The Italian experiences, positive and negative, of the post-earthquake are conditioned by long times and structural bureaucratic constraints, also motivated by the attempt to contain mafia infiltration and corruption. The transition from the operational phase of the emergency to the planning phase of the reconstruction project is thus hampered by a series of inefficiencies and delays, incompatible with the need for rapid recovery of the territories in crisis. In fact, intervening in areas affected by seismic events means at the same time associating the reconstruction plan with an urban and territorial rehabilitation project based on strategies and tools in which prevention and safety play a leading role in the regeneration of territories in crisis and the return of the population. On the contrary, the earthquakes that took place in Italy have instead further deprived the territories affected of the minimum requirements for habitability, in terms of accessibility and services, accentuating the depopulation process, already underway before the earthquake. The objective of this work is to address with implementing and programmatic tools the procedures and strategies to be put in place, today and in the future, in Italy and abroad, to face the challenge of the reconstruction of activities, sociality, services, risk mitigation: a protocol of operational intentions and firm points, open to a continuous updating and implementation. The methodology followed is that of the comparison in a synthetic form between the different Italian experiences of the post-earthquake, based on facts and not on intentions, to highlight elements of excellence or, on the contrary, damage. The main results obtained can be summarized in technical comparison cards on good and bad practices. With this comparison, we intend to make a concrete contribution to the reconstruction process, certainly not only related to the reconstruction of buildings but privileging the primary social and economic needs. In this context, the recent instrument applied in Italy of the strategic urban and territorial SUM (Minimal Urban Structure) and the strategic monitoring process become dynamic tools for supporting reconstruction. The conclusions establish, by points, a protocol of interventions, the priorities for integrated socio-economic strategies, multisectoral and multicultural, and highlight the innovative aspects of 'inversion' of priorities in the reconstruction process, favoring the take-off of 'accelerator' interventions social and economic and a more updated system of coexistence with risks. In this perspective, reconstruction as a necessary response to the calamitous event can and must become a unique opportunity to raise the level of protection from risks and rehabilitation and development of the most fragile places in Italy and abroad.

Keywords: an operational protocol for reconstruction, operational priorities for coexistence with seismic risk, social and economic interventions accelerators of building reconstruction, the difficult post-earthquake reconstruction in Italy

Procedia PDF Downloads 126

Authors: V. Moser-Siegmeth, M. C. Gambal, M. Jelovcak, B. Prytek, I. Swietalsky, D. Würzl, C. Fida, V. Mühlegger

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Adaptability is the capacity to adjust without great difficulty to changing circumstances. Within our project, we aimed to detect whether older people living within a long-term care hospital lose the ability to adapt. Theoretical concepts are contradictory in their statements. There is also lack of evidence in the literature how the adaptability of older people changes over the time. Following research questions were generated: Are older residents of a long-term care facility able to adapt to changes within their daily routine? How long does it take for older people to adapt? The study was designed as a convergent parallel mixed method intervention study, carried out within a four-month period and took place within seven wards of a long-term care hospital. As a planned intervention, a change of meal-times was established. The inhabitants were surveyed with qualitative interviews and quantitative questionnaires and diaries before, during and after the intervention. In addition, a survey of the nursing staff was carried out in order to detect changes of the people they care for and how long it took them to adapt. Quantitative data was analysed with SPSS, qualitative data with a summarizing content analysis. The average age of the involved residents was 82 years, the average length of stay 45 months. The adaptation to new situations does not cause problems for older residents. 47% of the residents state that their everyday life has not changed by changing the meal times. 24% indicate ‘neither nor’ and only 18% respond that their daily life has changed considerably due to the changeover. The diaries of the residents, which were conducted over the entire period of investigation showed no changes with regard to increased or reduced activity. With regard to sleep quality, assessed with the Pittsburgh sleep quality index, there is little change in sleep behaviour compared to the two survey periods (pre-phase to follow-up phase) in the cross-table. The subjective sleep quality of the residents is not affected. The nursing staff points out that, with good information in advance, changes are not a problem. The ability to adapt to changes does not deteriorate with age or by moving into a long-term care facility. It only takes a few days to get used to new situations. This can be confirmed by the nursing staff. Although there are different determinants like the health status that might make an adjustment to new situations more difficult. In connection with the limitations, the small sample size of the quantitative data collection must be emphasized. Furthermore, the extent to which the quantitative and qualitative sample represents the total population, since only residents without cognitive impairments of selected units participated. The majority of the residents has cognitive impairments. It is important to discuss whether and how well the diary method is suitable for older people to examine their daily structure.

Keywords: adaptability, intervention study, mixed methods, nursing home residents

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Authors: Nadarajah I. Ramesh

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Much of the research on stochastic point process models for rainfall has focused on Poisson cluster models constructed from either the Neyman-Scott or Bartlett-Lewis processes. The doubly stochastic Poisson process provides a rich class of point process models, especially for fine-scale rainfall modelling. This paper provides an account of recent development on this topic and presents the results based on some of the fine-scale rainfall models constructed from this class of stochastic point processes. Amongst the literature on stochastic models for rainfall, greater emphasis has been placed on modelling rainfall data recorded at hourly or daily aggregation levels. Stochastic models for sub-hourly rainfall are equally important, as there is a need to reproduce rainfall time series at fine temporal resolutions in some hydrological applications. For example, the study of climate change impacts on hydrology and water management initiatives requires the availability of data at fine temporal resolutions. One approach to generating such rainfall data relies on the combination of an hourly stochastic rainfall simulator, together with a disaggregator making use of downscaling techniques. Recent work on this topic adopted a different approach by developing specialist stochastic point process models for fine-scale rainfall aimed at generating synthetic precipitation time series directly from the proposed stochastic model. One strand of this approach focused on developing a class of doubly stochastic Poisson process (DSPP) models for fine-scale rainfall to analyse data collected in the form of rainfall bucket tip time series. In this context, the arrival pattern of rain gauge bucket tip times N(t) is viewed as a DSPP whose rate of occurrence varies according to an unobserved finite state irreducible Markov process X(t). Since the likelihood function of this process can be obtained, by conditioning on the underlying Markov process X(t), the models were fitted with maximum likelihood methods. The proposed models were applied directly to the raw data collected by tipping-bucket rain gauges, thus avoiding the need to convert tip-times to rainfall depths prior to fitting the models. One advantage of this approach was that the use of maximum likelihood methods enables a more straightforward estimation of parameter uncertainty and comparison of sub-models of interest. Another strand of this approach employed the DSPP model for the arrivals of rain cells and attached a pulse or a cluster of pulses to each rain cell. Different mechanisms for the pattern of the pulse process were used to construct variants of this model. We present the results of these models when they were fitted to hourly and sub-hourly rainfall data. The results of our analysis suggest that the proposed class of stochastic models is capable of reproducing the fine-scale structure of the rainfall process, and hence provides a useful tool in hydrological modelling.

Keywords: fine-scale rainfall, maximum likelihood, point process, stochastic model

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Authors: Nidhi Prabha

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Transportation as a sector is one of the most significant infrastructural elements of the ‘urban.' The distinctness of an urban life in a city is marked by the dynamic movements that it enables within the city-space. Therefore it is important to study the public-transport systems that enable and foster mobility which characterizes the urban. It is also crucial to underscore the way one is examining the urban transport systems - either as an infrastructural unit in a strict physical-structural sense or as a structural unit which acts as a prism refracting multiple experiences depending on the location of the ‘commuter.' In the proposed paper, the attempt is to uncover and investigate the assumption of the neuter-commuter by looking at urban transportation in the secondary sense i.e. as a structural unit which is experienced differently by different kinds of commuters, thus making transportation deeply distributed with various social structures and locations like class or gender which map onto the transport systems. To this end, the public-transit systems operating in Urban Delhi i.e. the Delhi Metros and the Delhi Transport Corporation run public-buses are looked at as case studies. The study is premised on the knowledge and data gained from both primary and secondary sources. Primary sources include data and knowledge collected from fieldwork, the methodology for which has ranged from adopting ‘mixed-methods’ which is ‘Qualitative-then-Quantitative’ as well as borrowing ethnographic techniques. Apart from fieldwork, other primary sources looked at including Annual Reports and policy documents of the Delhi Metro Rail Corporation (DMRC) and the Delhi Transport Corporation (DTC), Union and Delhi budgets, Economic Survey of Delhi, press releases, etc. Secondary sources include the vast array of literature available on the critical nodes that inform the research like gender, transport geographies, urban-space, etc. The study indicates a deeply-distributed urban transport system wherein the various social-structural locations or different kinds of commuters map onto the way these different commuters experience mobility or movement within the city space. Mobility or movement, therefore, becomes gendered or has class-based ramifications. The neuter-commuter assumption is thus challenged. Such an understanding enables us to challenge the anonymity which the ‘urban’ otherwise claims it provides over the rural. The rural is opposed to the urban wherein urban ushers a modern way of life, breaking ties of traditional social identities. A careful study of the transport systems through the traveling patterns and choices of the commuters, however, indicate that this does not hold true as even the same ‘public-space’ of the transport systems allocates different places to different kinds of commuters. The central argument made though the research done is therefore that infrastructure like urban-transport-systems has to be studied and examined as seen beyond just a physical structure. The various experiences of daily mobility of different kinds of commuters have to be taken into account in order to design and plan more inclusive transport systems.

Keywords: gender, infrastructure, mobility, urban-transport-systems

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Authors: Rafał Szmajser, Katarzyna Świetla, Mariusz Andrzejewski

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Robotic process automation (RPA) is a technology of repeatable business processes performed using computer programs, robots that simulate the work of a human being. This approach assumes replacing an existing employee with the use of dedicated software (software robots) to support activities, primarily repeated and uncomplicated, characterized by a low number of exceptions. RPA application is widespread in modern business services, particularly in the areas of Finance, Accounting and Human Resources Management. By utilizing this technology, the effectiveness of operations increases while reducing workload, minimizing possible errors in the process, and as a result, bringing measurable decrease in the cost of providing services. Regardless of how the use of modern information technology is assessed, there are also some doubts as to whether we should replace human activities in the implementation of the automation in business processes. After the initial awe for the new technological concept, a reflection arises: to what extent does the implementation of RPA increase the efficiency of operations or is there a Business Case for implementing it? If the business case is beneficial, in which business processes is the greatest potential for RPA? A closer look at these issues was provided by in this research during which the respondents’ view of the perceived advantages resulting from the use of robotization and automation in financial and accounting processes was verified. As a result of an online survey addressed to over 500 respondents from international companies, 162 complete answers were returned from the most important types of organizations in the modern business services industry, i.e. Business or IT Process Outsourcing (BPO/ITO), Shared Service Centers (SSC), Consulting/Advisory and their customers. Answers were provided by representatives of the positions in their organizations: Members of the Board, Directors, Managers and Experts/Specialists. The structure of the survey allowed the respondents to supplement the survey with additional comments and observations. The results formed the basis for the creation of a business case calculating tangible benefits associated with the implementation of automation in the selected financial processes. The results of the statistical analyses carried out with regard to revenue growth confirmed the correctness of the hypothesis that there is a correlation between job position and the perception of the impact of RPA implementation on individual benefits. Second hypothesis (H2) that: There is a relationship between the kind of company in the business services industry and the reception of the impact of RPA on individual benefits was thus not confirmed. Based results of survey authors performed simulation of business case for implementation of RPA in selected Finance and Accounting Processes. Calculated payback period was diametrically different ranging from 2 months for the Account Payables process with 75% savings and in the extreme case for the process Taxes implementation and maintenance costs exceed the savings resulting from the use of the robot.

Keywords: automation, outsourcing, business process automation, process automation, robotic process automation, RPA, RPA business case, RPA benefits

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Authors: D. Bianco, A. Sollazzo, M. Barbarino, G. Elia, A. Smoraldi, N. Favaloro

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The activities, described in the present paper, have been conducted in the framework of the HYPROB-New Program, carried out by the Italian Aerospace Research Centre (CIRA) promoted and funded by the Italian Ministry of University and Research (MIUR) in order to improve the National background on rocket engine systems for space applications. The Program has the strategic objective to improve National system and technology capabilities in the field of liquid rocket engines (LRE) for future Space Propulsion Systems applications, with specific regard to LOX/LCH4 technology. The main purpose of the HYPROB program is to design and build a Propulsion Test Facility (HIMP) allowing test activities on Liquid Thrusters. The development of skills in liquid rocket propulsion can only pass through extensive test campaign. Following its mission, CIRA has planned the development of new testing facilities and infrastructures for space propulsion characterized by adequate sizes and instrumentation. The IMP test cell is devoted to testing articles representative of small combustion chambers, fed with oxygen and methane, both in liquid and gaseous phase. This article describes the activities that have been carried out for the evaluation of the acoustic impact, and its consequent mitigation. The impact of the simulated acoustic disturbance has been evaluated, first, using an approximated method based on experimental data by Baumann and Coney, included in “Noise and Vibration Control Engineering” edited by Vér and Beranek. This methodology, used to evaluate the free-field radiation of jet in ideal acoustical medium, analyzes in details the jet noise and assumes sources acting at the same time. It considers as principal radiation sources the jet mixing noise, caused by the turbulent mixing of jet gas and the ambient medium. Empirical models, allowing a direct calculation of the Sound Pressure Level, are commonly used for rocket noise simulation. The model named after K. Eldred is probably one of the most exploited in this area. In this paper, an improvement of the Eldred Standard model has been used for a detailed investigation of the acoustical impact of the Hyprob facility. This new formulation contains an explicit expression for the acoustic pressure of each equivalent noise source, in terms of amplitude and phase, allowing the investigation of the sources correlation effects and their propagation through wave equations. In order to enhance the evaluation of the facility acoustic impact, including an assessment of the mitigation strategies to be set in place, a more advanced simulation campaign has been conducted using both an in-house code for noise propagation and scattering, and a commercial code for industrial noise environmental impact, CadnaA. The noise prediction obtained with the revised Eldred-based model has then been used for formulating an empirical/BEM (Boundary Element Method) hybrid approach allowing the evaluation of the barrier mitigation effect, at the design. This approach has been compared with the analogous empirical/ray-acoustics approach, implemented within CadnaA using a customized definition of sources and directivity factor. The resulting impact evaluation study is reported here, along with the design-level barrier optimization for noise mitigation.

Keywords: acoustic impact, industrial noise, mitigation, rocket noise

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Authors: Susan Evans, Janna Gordon-Elliott, Katarzyna Wyka, Virginia Mutch

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During the rise of the COVID-19 pandemic, healthcare workers needed an intervention that could address their profound acute stress. Mindfulness-based stress reduction (MBSR) is a program that has long established effectiveness for mental and physical health outcomes. In recent years, MBSR has been modified such that the duration of both class time and number of sessions has been abbreviated, and its delivery has been adapted for online dissemination, thus increasing the likelihood that individuals who could most benefit from the program would do so. We sought to investigate whether a brief, online version of MBSR could be feasible and acceptable for health care workers (HCW) in acute stress in response to the COVID-19 pandemic. Participants were recruited via an email sent to all hospital employees, which spans residents, physicians, nurses, housekeeping, lab technicians, administrators, and others. Participating HCW were asked about their previous experience with mindfulness and asked to commit to a minimum of 3 sessions. They were then provided with four weekly 1-hour sessions online that included the major mindfulness exercises taught during traditional MBSR programs (i.e., body scan, sitting meditation, mindful eating, and yoga). Participants were provided with supporting slides, videos, demonstrations and asked to track their practice. Hospital staff enrolled in the program; by the end of the first day of recruitment, 40 had applied; by the start date, about 100 were enrolled, and n attended a minimum of 3 sessions, supporting feasibility. Hospital staff also participated and practiced the mindfulness exercises (n=42), thus supporting acceptability. Participants reported that the program was logical, successful, and worth recommending both before starting the program and after completing it (M= 22.02 and M=21.76, respectively, possible range 0-27). There was a slight decline in the belief in improvement in health and well-being due to the program (ES=.37, p=.021). Secondary hypotheses regarding participants’ self-reported stress and levels of mindfulness were also supported, such that participants reported improvements in perceived stress (ES=.45, p=.006), compassion satisfaction, burnout, and secondary traumatic stress (ES=.41, ES=.31, ES=.35, respectively, p<.05). Participants reported significant improvements in the describing facet of mindfulness (ES=.49, p=.004), while all other facets (observing, acting with awareness, nonjudging of inner experience, nonreactivity to inner experience) remained unchanged pre- to post-program. Results from this study suggest that an abridged, online version of MBSR is feasible and accessible to health care workers in acute stress and provides benefits expected from traditional MBSR programs. The lack of a randomized control group limits generalizability. We intend to provide a structure, framework, and lessons learned to hospital administrators and clinical staff seeking to support their employees in acute stress.

Keywords: acute stress, health care workers, mindfulness, online interventions

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Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian

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Kawasaki disease (KD) is the primary cause of acquired pediatric heart disease as an acute vasculitis. In children with prolonged fever, rash, and inflammation of the mucosa KD must be considered as a clinical diagnosis. There is a persuasive suggestion of immune-mediated damage as the pathophysiologic cascade of KD. For example, the invasion of cytotoxic T-cells supports a viral etiology and the inflammasome of the innate immune system is a critical component in the vasculitis formation in KD. Animal models of KD propose the cytokine profiles, such as increased IL-1 and GM-CSF, which cause vascular damage. CRP and IFN-γ elevated expression and the upregulation of IL-6, and IL-10 production are also described in previous studies. Untreated KD is a critical risk factor for coronary artery diseases and myocardial infarction. Vascular damage may encompass amplified T-cell activity. SMAD3 is an essential molecule in down-regulating T-cells and increasing expression of FoxP3. It has a critical effect in the differentiation of regulatory T-cells. The discrepancy of regulatory T-cells and pro-inflammatory Th17 has been studied in acute coronary syndrome during KD. However in the coronary artery damaged lymphocytes and IgA plasma cells are seen at the lesion locations, the major immune cells in the coronary lesions are monocytes/macrophages and neutrophils. These cells secrete TNF-α, and activates matrix metalloprotease (MMP)-9, reducing the integrity of vessels and prompting patients to arise aneurysm. MMPs can break down the components of the extracellular matrix and assist immune cell movement. IVIG as an effective form of treatment clarified the role of the immune system, which may target pathogenic antigens and regulate cytokine production. Several reports have revealed that in the coronary arteries, high expression of MMP-9 in monocyte/macrophage results in pathologic cascades. Curcumin is a potent antioxidant and anti-inflammatory molecule. Curcumin decreases the production of reactive oxygen and nitrogen species and inhibits transcription factors like AP-1 and NF-κB. Curcumin also contains the characteristics of inhibitory effects on MMPs, especially MMP-9. The upregulation of MMP-9 is an important cellular response. Curcumin treatment caused a reverse effect and down-regulates MMP-9 gene expression which may fund the anti-inflammatory effect. Curcumin inhibits MMP-9 expression via PKC and AMPK-dependent pathways in Human monocytes cells. Elevated expression and activity of MMP-9 are correlated with advanced vascular lesions. AMPK controls lipid metabolism and oxidation, and protein synthesis. AMPK is also necessary for the MMP-9 activity and THP-1 cell adhesion to endothelial cells. Curcumin was shown to inhibit the activation of AMPKα. Compound C (AMPK inhibitor) inhibits MMP-9 expression level. Therefore, through inactivating AMPKs and PKC, curcumin decreases the MMP-9 level, which results in inhibiting monocyte/macrophage differentiation. Compound C also suppress the phosphorylation of three major classes of MAP kinase signaling, suggesting that curcumin may suppress MMP-9 level by inactivation of MAPK pathways. MAPK cascades are activated to induce the expression of MMP-9. Curcumin inhibits MAPKs phosphorylation, which contributes to the down-regulation of MMP-9. This study demonstrated that the potential inhibitory properties of curcumin over MMP-9 lead to a therapeutic strategy to reduce the risk of coronary artery involvement during KD.

Keywords: MMP-9, coronary artery aneurysm, Kawasaki’s disease, curcumin, AMPK, immune system, NF-κB, MAPK

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Authors: Joanna Gerszon, Aleksandra Rodacka

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In the pathogenesis of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, protein and peptide aggregation processes play a vital role in contributing to the formation of intracellular and extracellular protein deposits. One of the major components of these deposits is the oxidatively modified glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Therefore, the purpose of this research was to answer the question whether piceatannol, a stilbene derivative, counteracts and/or slows down oxidative stress-induced GAPDH aggregation. The study also aimed to determine if this natural occurring compound prevents unfavorable nuclear translocation of GAPDH in hippocampal cells. The isothermal titration calorimetry (ITC) analysis indicated that one molecule of GAPDH can bind up to 8 molecules of piceatannol (7.3 ± 0.9). As a consequence of piceatannol binding to the enzyme, the loss of activity was observed. Parallel with GAPDH inactivation the changes in zeta potential, and loss of free thiol groups were noted. Nevertheless, the ligand-protein binding does not influence the secondary structure of the GAPDH. Precise molecular docking analysis of the interactions inside the active center allowed to presume that these effects are due to piceatannol ability to assemble a covalent binding with nucleophilic cysteine residue (Cys149) which is directly involved in the catalytic reaction. Molecular docking also showed that simultaneously 11 molecules of ligand can be bound to dehydrogenase. Taking into consideration obtained data, the influence of piceatannol on level of GAPDH aggregation induced by excessive oxidative stress was examined. The applied methods (thioflavin-T binding-dependent fluorescence as well as microscopy methods - transmission electron microscopy, Congo Red staining) revealed that piceatannol significantly diminishes level of GAPDH aggregation. Finally, studies involving cellular model (Western blot analyses of nuclear and cytosolic fractions and confocal microscopy) indicated that piceatannol-GAPDH binding prevents GAPDH from nuclear translocation induced by excessive oxidative stress in hippocampal cells. In consequence, it counteracts cell apoptosis. These studies demonstrate that by binding with GAPDH, piceatannol blocks cysteine residue and counteracts its oxidative modifications, that induce oligomerization and GAPDH aggregation as well as it prevents hippocampal cells from apoptosis by retaining GAPDH in the cytoplasm. All these findings provide a new insight into the role of piceatannol interaction with GAPDH and present a potential therapeutic strategy for some neurological disorders related to GAPDH aggregation. This work was supported by the by National Science Centre, Poland (grant number 2017/25/N/NZ1/02849).

Keywords: glyceraldehyde-3-phosphate dehydrogenase, neurodegenerative disease, neuroprotection, piceatannol, protein aggregation

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Authors: Marco Ranzato, Maguelone Vignes

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The co-design process recently run by the trans-disciplinary urban laboratory Metrolab Brussels for outlining the architecture of a future integrated health centre in Brussels (Belgium) has highlighted that a buffer place open to the local community is the appropriate cornerstone around which organizing a space where diverse professionals and patients are together. In the context of the migrants 'crisis' in Europe, the growing number of vulnerable people in Brussels and the increasing complexity of the health and welfare systems, the NGO Doctors of the World (DoW) has launched a project funded by The European Regional Development Fund, and aiming to create a new community centre combining social and health services in a poor but changing neighborhood of Brussels. Willing not to make a 'ghetto' of this new integrated service, the NGO looks at hosting different publics in order to make the poorest, marginal and most vulnerable people access to a regular kind of service. As a trans-disciplinary urban research group, Metrolab has been involved in the process of co-designing the architecture of the future centre with a set of various health professionals, social workers, and patients’ representatives. Metrolab drawn on the participants’ practice experiences and knowledge of hosting different kinds of publics and professions in a same structure in order to imagine what rooms should fit into the centre, what atmosphere they should convey, how should they be interrelated and organized, and, concurrently, how the building should fit into the urban frame of its neighborhood. The result is that, in order for an integrated health centre framed in the landscape of a disadvantaged neighborhood to function, it has to work as social community centre offering accessibility and conviviality to diverse social groups. This paper outlines the methodology that Metrolab used to design and conduct, in close collaboration with DoW, a series of 3 workshops. Through sketching and paper modeling, the methodology made participants talk about their experience by projecting them into a situation. It included a combination of individual and collective work in order to sharp participants’ eyes on architectural forms, explicit their thoughts and experience through inter-subjectivity and imagine solutions to the challenges they raised. Such a collaborative method encompasses several challenges about patients’ participation and representation, replicability of the conditions of success and the plurality of the research findings communication formats. This paper underlines how this participatory process has contributed to build knowledge on the few-documented topic of the architecture of community health centres. More importantly, the contribution builds on this participatory process to discuss the importance of adapting the architecture of the new integrated health centre to the changing population of Brussels and to the issues of its specific neighborhood.

Keywords: co-design, health, social innovation, urban lab

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Authors: Asghar Ali, Narjis Khatoon

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This study focuses on the presence of wage differentials in Pakistan and also on the determinants that originate it. Since there are a smaller number of studies that are conducted on this topic in Pakistan, the current study aims to contribute in bridging the existing gap in this particular research genre. Hence, this study not only generates the desired results specific focus but it also contributes to the overall empirical work on the Pakistan economy. The preceding works which have been done to research wage determinants and wage differentials have used numerous different theories and approaches to reach their goals. The current study, in order to analyze the determinants of wage differentials in the developing economy, deals with the study of a number of such theories and approaches that are supposed as being beneficial for the purpose. This study undertakes the explanation of wage differentials in Pakistan by focusing on wage differentials in public and private sectors, formal and informal sectors, and major occupational groups. The study uses 'Wage Theory' to examine wage differentials among male and female employees in public and private sectors on varied levels of working conditions. This study also uses 'Segmented Labor Market Theory' to determine the wage differential in both public and private sectors, formal and informal, and major occupational groups in Pakistan. So the author has used various econometric techniques in order to explain and test these theories and to find out the required results. This study has employed seven different cross-sectional Labour Force Surveys for the time period between 2006-07 to 2012-13. Gender equality is not only a policy reform agenda for developing countries but also an important goal of Millennium Development Goals. This study investigates the nexus between wage inequality and economic growth and detects co-integration between gender wage differential and economic growth using ARDL bound test. It is confirmed from the empirical results that there exists long-run relationship between economic growth and wage differential. Our study indicated that half of the total female employees from fourteen major cities of Pakistan were employed in the public sector. Out of total female employees in private sector, 66 percent are employed in the formal sector, and 33 percent are working in the informal sector. Results also indicated that both men and women were paid more in the public sector compared to the private sector counterparts. Among the total female employees, only 9 percent had received any formal training, 52% were married and average years of schooling were 11 years. Further, our findings regarding wage differential between genders indicate that wage gap is lower in public sector as compared to private sector. In proportion, gender wage ratio was found to be 0.96, 0.62 and 0.66 in public, formal private and informal private sectors respectively. This suggests that in this case, private sector female employees with the same pay structure are compensated at a lower endowments rate as then public sector workers as compared to their counter parts.

Keywords: wage differentials, formal, informal, economic growth

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Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

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Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

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