Search results for: heat shock protein expression

Authors: Ceren Ince, Berkay Zafer Erdem, Shahram Derogar, Nabi Yuzer

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Fibre cement plates, often used in construction, generally are made using quartz as an inert material, cement as a binder and cellulose as a fibre. This paper first of all investigates the mechanical properties and durability of fibre cement plates when quartz is both partly and fully replaced with diatomite. Diatomite does not only have lower density compared to quartz but also has high pozzolanic activity. The main objective of this paper is the investigation of the effects of supplementary cementing materials (SCMs) on the short and long term mechanical properties and durability characteristics of fibre cement plates prepared using diatomite. Supplementary cementing materials such as ground granulated blast furnace slug (GGBS) and fly ash (FA) are used in this study. 10, 20, 30 and 40% of GGBS and FA are used as partial replacement materials to cement. Short and long term mechanical properties such as compressive and flexural strengths as well as capillary absorption, sorptivity characteristics and mass were investigated. Consistency and setting time at each replacement levels of SCMs were also recorded. The effects of using supplementary cementing materials on the carbonation and sulphate resistance of fibre cement plates were then experimented. The results, first of all, show that the use of diatomite as a full or partial replacement to quartz resulted in a systematic decrease in total mass of the fibre cement plates. The reduction of mass was largely due to the lower density and finer particle size of diatomite compared to quartz. The use of diatomite did not only reduce the mass of these plates but also increased the compressive strength significantly as a result of its high pozzolanic activity. The replacement levels of both GGBS and FA resulted in a systematic decrease in short term compressive strength with increasing replacement levels. This was essentially expected as the total heat of hydration is much lower in GGBS and FA than that of cement. Long term results however, indicated that the compressive strength of fibre cement plates prepared using both GGBS and FA increases with time and hence the compressive strength of plates prepared using SCMs is either equivalent or more than the compressive strength of plates prepared using cement alone. Durability characteristics of fibre cement plates prepared using SCMs were enhanced significantly. Measurements of capillary absorption and sopritivty characteristics were also indicated that the plates prepared using SCMs has much lower permeability compared to plates prepared cement alone. Much higher resistance to carbonation and sulphate attach were observed with plates prepared using SCMs. The results presented in this paper show that the use of SCMs does not only support the production of more sustainable construction materials but also enhances the mechanical properties and durability characteristics of fibre cement plates.

Keywords: diatomite, fibre, strength, supplementary cementing material

Procedia PDF Downloads 317

Authors: N. Schneider, M. Bruchhage, M. Hartweg, G. Mutungi, J. O Regan, S. Deoni

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Observational neuroimaging studies suggest differences between breast-fed and formula-fed infants in developmental myelination, a key brain process for learning and cognitive development. However, the possible effects of a nutrient formulation on myelin development in healthy term infants in an intervention study have not been investigated. Objective was, therefore, to investigate the efficacy of a nutrient formulation with higher levels of myelin-relevant nutrients as compared to a control formulation with lower levels of the same nutrients on brain myelination and cognitive development in the first 6 months of life. The study is an ongoing randomized, controlled, double-blind, two-center, parallel-group clinical trial with a nonrandomized, non-blinded arm of exclusively breastfed infants. The current findings result from a staged statistical analysis at 6 months; the recruitment and intervention period has been completed for all participants. Follow-up visits at 12, 18 and 24 months are still ongoing. N= 81 enrolled full term, neurotypical infants of both sexes were randomized into either the investigational (N= 42) or the control group (N= 39), and N= 108 children in the breast-fed arm served as a natural reference group. The effect of a blend of docosahexaenoic acid, arachidonic acid, iron, vitamin B12, folic acid as well as sphingomyelin from a uniquely proceed whey protein concentrate enriched in alpha-lactalbumin and phospholipids in an infant nutrition product matrix was investigated. The main outcomes for the staged statistical analyses at 6 months included brain myelination measures derived from MRI. Additional outcomes were brain volume, cognitive development and safety. The full analyses set at 6 months comprised N= 66 infants. Higher levels of myelin-relevant nutrients compared to lower levels resulted in significant differences in myelin structure, volume, and rate of myelination as early as 3 and 6 months of life. The cross-sectional change of means between groups for whole-brain myelin volume was 8.4% for investigational versus control formulation (3.5% versus the breastfeeding reference) group at 3 months and increased to 36.4% for investigational versus control formulation (14.1% versus breastfeeding reference) at 6 months. No statistically significant differences were detected for early cognition scores. Safety findings were largely similar across groups. This is the first pediatric nutritional neuroimaging study demonstrating the efficacy of a myelin nutrient blend on developmental myelination in well-nourished term infants. Myelination is a critical process in learning and development. The effects were demonstrated across the brain, particularly in temporal and parietal regions, known to be functionally involved in sensory, motor and language skills. These first results add to the field of nutritional neuroscience by demonstrating early life nutrition benefits for brain architecture which may be foundational for later cognitive and behavioral outcomes. ClinicalTrials.gov Identifier: NCT03111927 (Infant Nutrition and Brain Development - Full-Text View - ClinicalTrials.gov).

Keywords: brain development, infant nutrition, MRI, myelination

Procedia PDF Downloads 177

Authors: H. Jamili, S. Shakiba

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Today, one of the main concerns of human beings is facing the unpleasant changes of the environment. Buildings are responsible for a significant amount of natural resources consumption and carbon emissions production. In such a situation, this thought comes to mind that changing each building into a phenomenon of benefit to the environment. A change in a way that each building functions as an element that supports the environment, and construction, in addition to answering the need of humans, is encouraged, the way planting a tree is, and it is no longer seen as a threat to alive beings and the planet. Prospect: Today, different ideas of developing materials that can smartly function are realizing. For instance, Programmable Materials, which in different conditions, can respond appropriately to the situation and have features of modification in shape, size, physical properties and restoration, and repair quality. Studies are to progress having this purpose to plan for these materials in a way that they are easily available, and to meet this aim, there is no need to use expensive materials and high technologies. In these cases, physical attributes of materials undertake the role of sensors, wires and actuators then materials will become into robots itself. In fact, we experience robotics without robots. In recent decades, AI and technology advances have dramatically improving the performance of materials. These achievements are a combination of software optimizations and physical productions such as multi-materials 3D printing. These capabilities enable us to program materials in order to change shape, appearance, and physical properties to interact with different situations. nIt is expected that further achievements like Memory Materials and Self-learning Materials are also added to the Smart Materials family, which are affordable, available, and of use for a variety of applications and industries. From the architectural standpoint, the building skin is significantly considered in this research, concerning the noticeable surface area the buildings skin have in urban space. The purpose of this research would be finding a way that the programmable materials be used in building skin with the aim of having an effective and positive interaction. A Pixel Façade would be a solution for programming a building skin. The Pixel Facadeincludes components that contain a series of attributes that help buildings for their needs upon their environmental criteria. A PIXEL contains series of smart materials and digital controllers together. It not only benefits its physical properties, such as control the amount of sunlight and heat, but it enhances building performance by providing a list of features, depending on situation criteria. The features will vary depending on locations and have a different function during the daytime and different seasons. The primary role of a PIXEL FAÇADE can be defined as filtering pollutions (for inside and outside of the buildings) and providing clean energy as well as interacting with other PIXEL FACADES to estimate better reactions.

Keywords: building skin, environmental crisis, pixel facade, programmable materials, smart materials

Procedia PDF Downloads 79

Authors: Sara A. Pakvis, Giulia Scalet, Stefania Marconi, Ferdinando Auricchio, Matthijs Langelaar

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In recent developments in the field of multi-material additive manufacturing, differences in material properties are exploited to create printed shape-memory structures, which are referred to as 4D-printed structures. New printing techniques allow for the deliberate introduction of prestresses in the specimen during manufacturing, and, in combination with the right design, this enables new functionalities. This research focuses on bi-polymer 4D-printed structures, where the transformation process is based on a heat-induced glass transition in one material lowering its Young’s modulus, combined with an initial prestress in the other material. Upon the decrease in stiffness, the prestress is released, which results in the realization of an essentially pre-programmed deformation. As the design of such functional multi-material structures is crucial but far from trivial, a systematic methodology to find the design of 4D-printed structures is developed, where a finite element model is combined with a density-based topology optimization method to describe the material layout. This modeling approach is verified by a convergence analysis and validated by comparing its numerical results to analytical and published data. Specific aspects that are addressed include the interplay between the definition of the prestress and the material interpolation function used in the density-based topology description, the inclusion of a temperature-dependent stiffness relationship to simulate the glass transition effect, and the importance of the consideration of geometric nonlinearity in the finite element modeling. The efficacy of topology optimization to design 4D-printed structures is explored by applying the methodology to a variety of design problems, both in 2D and 3D settings. Bi-layer designs composed of thermoplastic polymers are printed by means of the fused deposition modeling (FDM) technology. Acrylonitrile butadiene styrene (ABS) polymer undergoes the glass transition transformation, while polyurethane (TPU) polymer is prestressed by means of the 3D-printing process itself. Tests inducing shape transformation in the printed samples through heating are performed to calibrate the prestress and validate the modeling approach by comparing the numerical results to the experimental findings. Using the experimentally obtained prestress values, more complex designs have been generated through topology optimization, and samples have been printed and tested to evaluate their performance. This study demonstrates that by combining topology optimization and 4D-printing concepts, stimuli-responsive structures with specific properties can be designed and realized.

Keywords: 4D-printing, glass transition, shape memory polymer, topology optimization

Procedia PDF Downloads 182

Authors: Mohammad Nazri Mustafa, Sh Norliza Sy Salim, Pedram Hatami Abdullah

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Civil and structural assets normally have an average of more than 30 years of design life. Adding to this advantage, the assets are normally subjected to slow degradation process. Due to the fact that repair and strengthening work for these assets are normally not dependent on plant shut down, the maintenance and integrity restoration of these assets are mostly done based on “as required” and “run to failure” basis. However unlike other industries, the exposure in oil and gas environment is harsher as the result of corrosive soil and groundwater, chemical spill, frequent wetting and drying, icing and de-icing, steam and heat, etc. Due to this type of exposure and the increasing level of structural defects and rectification in line with the increasing age of plants, assets integrity assessment requires a more defined scope and procedures that needs to be based on risk and assets criticality. This leads to the establishment of risk based inspection and proactive maintenance procedure for civil and structural assets. To date there is hardly any procedure and guideline as far as integrity assessment and systematic inspection and maintenance of civil and structural assets (onshore) are concerned. Group Technical Solutions has developed a procedure and guideline that takes into consideration credible failure scenario, assets risk and criticality from process safety and structural engineering perspective, structural importance, modeling and analysis among others. Detailed inspection that includes destructive and non-destructive tests (DT & NDT) and structural monitoring is also being performed to quantify defects, assess severity and impact on integrity as well as identify the timeline for integrity restoration. Each defect and its credible failure scenario is assessed against the risk on people, environment, reputation and production loss. This technical paper is intended to share on the established procedure and guideline and their execution in oil & gas plants. In line with the overall roadmap, the procedure and guideline will form part of specialized solutions to increase production and to meet the “Operational Excellence” target while extending service life of civil and structural assets. As the result of implementation, the management of civil and structural assets is now more systematically done and the “fire-fighting” mode of maintenance is being gradually phased out and replaced by a proactive and preventive approach. This technical paper will also set the criteria and pose the challenge to the industry for innovative repair and strengthening methods for civil & structural assets in oil & gas environment, in line with safety, constructability and continuous modification and revamp of plant facilities to meet production demand.

Keywords: assets criticality, credible failure scenario, proactive and preventive maintenance, risk based inspection

Procedia PDF Downloads 381

Authors: Antoine Lauvray, Fabien Poulhaon, Pierre Michaud, Pierre Joyot, Emmanuel Duc

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Additive Friction Stir Manufacturing (AFSM) is a new industrial process that follows the emergence of friction-based processes. The AFSM process is a solid-state additive process using the energy produced by the friction at the interface between a rotating non-consumable tool and a substrate. Friction depends on various parameters like axial force, rotation speed or friction coefficient. The feeder material is a metallic rod that flows through a hole in the tool. Unlike in Friction Stir Welding (FSW) where abundant literature exists and addresses many aspects going from process implementation to characterization and modeling, there are still few research works focusing on AFSM. Therefore, there is still a lack of understanding of the physical phenomena taking place during the process. This research work aims at a better AFSM process understanding and implementation, thanks to numerical simulation and experimental validation performed on a prototype effector. Such an approach is considered a promising way for studying the influence of the process parameters and to finally identify a process window that seems relevant. The deposition of material through the AFSM process takes place in several phases. In chronological order these phases are the docking phase, the dwell time phase, the deposition phase, and the removal phase. The present work focuses on the dwell time phase that enables the temperature rise of the system composed of the tool, the filler material, and the substrate and due to pure friction. Analytic modeling of heat generation based on friction considers as main parameters the rotational speed and the contact pressure. Another parameter considered influential is the friction coefficient assumed to be variable due to the self-lubrication of the system with the rise in temperature or the materials in contact roughness smoothing over time. This study proposes, through numerical modeling followed by experimental validation, to question the influence of the various input parameters on the dwell time phase. Rotation speed, temperature, spindle torque, and axial force are the main monitored parameters during experimentations and serve as reference data for the calibration of the numerical model. This research shows that the geometry of the tool as well as fluctuations of the input parameters like axial force and rotational speed are very influential on the temperature reached and/or the time required to reach the targeted temperature. The main outcome is the prediction of a process window which is a key result for a more efficient process implementation.

Keywords: numerical model, additive manufacturing, friction, process

Procedia PDF Downloads 132

Authors: Sahar Yousef, Chantelle Niblock, Gul Kacmaz

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Salt City, in the context of Jordan’s heritage landscape, is a significant case to explore when it comes to the interaction between tangible and intangible qualities of liveable cities. Most city centers, including Jerash, Salt, Irbid, and Amman, are historical locations. Six of these extraordinary sites were designated UNESCO World Heritage Sites. Jordan is widely acknowledged as a developing country characterized by swift urbanization and unrestrained expansion that exacerbate the challenges associated with the preservation of historic urban areas. The aim of this study is to conduct an examination and analysis of the existing condition of heritage connectivity within heritage city centers. This includes outdoor staircases, pedestrian pathways, footpaths, and other public spaces. Case study-style analysis of the urban core of As-Salt is the focus of this investigation. Salt City is widely acknowledged for its substantial tangible and intangible cultural heritage and has been designated as ‘The Place of Tolerance and Urban Hospitality’ by UNESCO since 2021. Liveability in urban heritage, particularly in historic city centers, incorporates several factors that affect our well-being; its enhancement is a critical issue in contemporary society. The dynamic interaction between humans and historical materials, which serves as a vehicle for the expression of their identity and historical narrative, constitutes preservation that transcends simple conservation. This form of engagement enables people to appreciate the diversity of their heritage recognising their previous and planned futures. Heritage preservation is inextricably linked to a larger physical and emotional context; therefore, it is difficult to examine it in isolation. Urban environments, including roads, structures, and other infrastructure, are undergoing unprecedented physical design and construction requirements. Concurrently, heritage reinforces a sense of affiliation with a particular location or space and unifies individuals with their ancestry, thereby defining their identity. However, a considerable body of research has focused on the conservation of heritage buildings in a fragmented manner without considering their integration within a holistic urban context. Insufficient attention is given to the significance of the physical and social roles played by the heritage staircases and baths that serve as connectors between these valued historical buildings. In doing so, the research uses a methodology that is based on consensus. Given that liveability is considered a complex matter with several dimensions. The discussion starts by making initial observations on the physical context and societal norms inside the urban center while simultaneously establishing the definitions of liveability and connectivity and examining the key criteria associated with these concepts. Then, identify the key elements that contribute to liveable connectivity within the framework of urban heritage in Jordanian city centers. Some of the outcomes that will be discussed in the presentation are: (1) There is not enough connectivity between heritage buildings as can be seen, for example, between buildings in Jada and Qala'. (2) Most of the outdoor spaces suffer from physical issues that hinder their use by the public, like in Salalem. (3) Existing activities in the city center are not well attended because of lack of communication between the organisers and the citizens.

Keywords: connectivity, Jordan, liveability, salt city, tangible and intangible heritage, urban heritage

Procedia PDF Downloads 46

Authors: J. N. Nackler, K. Saleh Pascha, W. Winter

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WHISCERS™ (Whole House In-Situ Carbon and Energy Reduction Solution) is an innovative process for Internal Wall Insulation (IWI) for energy-efficient retrofitting of heritage building, which uses laser measuring to determine the dimensions of a room, off-site insulation board cutting and rapid installation to complete the process. As part of a multinational investigation consortium the Austrian part adapted the WHISCERS system to local conditions of Vienna where most historical buildings have valuable stucco facades, precluding the application of an external insulation. The Austrian project contribution addresses the replacement of commonly used extruded polystyrene foam (XPS) with renewable materials such as wood and wood products to develop a more sustainable IWI system. As the timber industry is a major industry in Austria, a new innovative and more sustainable IWI solution could also open up new markets. The first approach of investigation was the Life Cycle Assessment (LCA) to define the performance of wood fibre board as insulation material in comparison to normally used XPS-boards. As one of the results the global-warming potential (GWP) of wood-fibre-board is 15 times less the equivalent to carbon dioxide while in the case of XPS it´s 72 times more. The hygrothermal simulation program WUFI was used to evaluate and simulate heat and moisture transport in multi-layer building components of the developed IWI solution. The results of the simulations prove in examined boundary conditions of selected representative brickwork constructions to be functional and usable without risk regarding vapour diffusion and liquid transport in proposed IWI. In a further stage three different solutions were developed and tested (1 - glued/mortared, 2 - with soft board, connected to wall with gypsum board as top layer, 3 - with soft board and clay board as top layer). All three solutions presents a flexible insulation layer out of wood fibre towards the existing wall, thus compensating irregularities of the wall surface. From first considerations at the beginning of the development phase, three different systems had been developed and optimized according to assembly technology and tested as small specimen in real object conditions. The built prototypes are monitored to detect performance and building physics problems and to validate the results of the computer simulation model. This paper illustrates the development and application of the Internal Wall Insulation system.

Keywords: internal insulation, wood fibre, hygrothermal simulations, monitoring, clay, condensate

Procedia PDF Downloads 203

Authors: Ahmed Borg, Elsa Aristodemou, Attia Attia

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Well control involves managing the circulating drilling fluid within the wells and avoiding kicks and blowouts as these can lead to losses in human life and drilling facilities. Current practices for good control incorporate predictions of pressure losses through computational models. Developing a realistic hydraulic model for a good control problem is a very complicated process due to the existence of a complex multiphase region, which usually contains a non-Newtonian drilling fluid and the miscibility of formation gas in drilling fluid. The current approaches assume an inaccurate flow fluid model within the well, which leads to incorrect pressure loss calculations. To overcome this problem, researchers have been considering the more complex two-phase fluid flow models. However, even these more sophisticated two-phase models are unsuitable for applications where pressure dynamics are important, such as in managed pressure drilling. This study aims to develop and implement new fluid flow models that take into consideration the miscibility of fluids as well as their non-Newtonian properties for enabling realistic kick treatment. furthermore, a corresponding numerical solution method is built with an enriched data bank. The research work considers and implements models that take into consideration the effect of two phases in kick treatment for well control in conventional drilling. In this work, a corresponding numerical solution method is built with an enriched data bank. Software STARCCM+ for the computational studies to study the important parameters to describe wellbore multiphase flow, the mass flow rate, volumetric fraction, and velocity of each phase. Results showed that based on the analysis of these simulation studies, a coarser full-scale model of the wellbore, including chemical modeling established. The focus of the investigations was put on the near drill bit section. This inflow area shows certain characteristics that are dominated by the inflow conditions of the gas as well as by the configuration of the mud stream entering the annulus. Without considering the gas solubility effect, the bottom hole pressure could be underestimated by 4.2%, while the bottom hole temperature is overestimated by 3.2%. and without considering the heat transfer effect, the bottom hole pressure could be overestimated by 11.4% under steady flow conditions. Besides, larger reservoir pressure leads to a larger gas fraction in the wellbore. However, reservoir pressure has a minor effect on the steady wellbore temperature. Also as choke pressure increases, less gas will exist in the annulus in the form of free gas.

Keywords: multiphase flow, well- control, STARCCM+, petroleum engineering and gas technology, computational fluid dynamic

Procedia PDF Downloads 100

Authors: Qian Li, Zhaoping Zhong

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Sewage sludge, a typical solid waste, has inevitably been produced in enormous quantities in China. Still worse, the amount of sewage sludge produced has been increasing due to rapid economic development and urbanization. Compared to the conventional method to treat sewage sludge, pyrolysis has been considered an economic and ecological technology because it can significantly reduce the sludge volume, completely kill pathogens, and produce valuable solid, gas, and liquid products. However, the large-scale utilization of sludge biochar has been limited due to the considerable risk posed by heavy metals in the sludge. Heavy metals enriched in pyrolytic biochar could be divided into exchangeable, reducible, oxidizable, and residual forms. The residual form of heavy metals is the most stable and cannot be used by organisms. Kaolin and zeolite are environmentally friendly inorganic minerals with a high surface area and heat resistance characteristics. So, they exhibit the enormous potential to immobilize heavy metals. In order to reduce the risk of leaching heavy metals in the pyrolysis biochar, this study pyrolyzed sewage sludge mixed with kaolin/zeolite in a small rotary kiln. The influences of additives and pyrolysis temperature on the leaching concentration and morphological transformation of heavy metals in pyrolysis biochar were investigated. The potential mechanism of stabilizing heavy metals in the co-pyrolysis of sludge blended with kaolin/zeolite was explained by scanning electron microscopy, X-ray diffraction, and specific surface area and porosity analysis. The European Community Bureau of Reference sequential extraction procedure has been applied to analyze the forms of heavy metals in sludge and pyrolysis biochar. All the concentrations of heavy metals were examined by flame atomic absorption spectrophotometry. Compared with the proportions of heavy metals associated with the F4 fraction in pyrolytic carbon prepared without additional agents, those in carbon obtained by co-pyrolysis of sludge and kaolin/zeolite increased. Increasing the additive dosage could improve the proportions of the stable fraction of various heavy metals in biochar. Kaolin exhibited a better effect on stabilizing heavy metals than zeolite. Aluminosilicate additives with excellent adsorption performance could capture more released heavy metals during sludge pyrolysis. Then heavy metal ions would react with the oxygen ions of additives to form silicate and aluminate, causing the conversion of heavy metals from unstable fractions (sulfate, chloride, etc.) to stable fractions (silicate, aluminate, etc.). This study reveals that the efficiency of stabilizing heavy metals depends on the formation of stable mineral compounds containing heavy metals in pyrolysis biochar.

Keywords: co-pyrolysis, heavy metals, immobilization mechanism, sewage sludge

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Authors: Andrés Lara Guillén, José M. Soriano Disla, Gemma Castejón Martínez, David Fernández-Gutiérrez

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The worldwide population is exponentially increasing, which causes a rising demand for food, energy and non-renewable resources. These demands must be attended to from a circular economy point of view. Under this approach, the obtention of strategic products from biowaste is crucial for the society to keep the current lifestyle reducing the environmental and social issues linked to the lineal economy. This is the main objective of the VALUEWASTE project. VALUEWASTE is about valorizing urban biowaste into proteins for food and feed and biofertilizers, closing the loop of this waste stream. In order to achieve this objective, the project validates three value chains, which begin with the anaerobic digestion of the biowaste. From the anaerobic digestion, three by-products are obtained: i) methane that is used by microorganisms, which will be transformed into microbial proteins; ii) digestate that is used by black soldier fly, producing insect proteins; and iii) a nutrient-rich effluent, which will be transformed into biofertilizers. VALUEWASTE is an innovative solution, which combines different technologies to valorize entirely the biowaste. However, it is also required to demonstrate that the solution is greener than other traditional technologies (baseline systems). On one hand, the proteins from microorganisms and insects will be compared with other reference protein production systems (gluten, whey and soybean). On the other hand, the biofertilizers will be compared to the production of mineral fertilizers (ammonium sulphate and synthetic struvite). Therefore, the aim of this study is to provide that biowaste valorization can reduce the environmental impacts linked to both traditional proteins manufacturing processes and mineral fertilizers, not only at a pilot-scale but also at an industrial one. In the present study, both baseline system and VALUEWASTE solution are evaluated through the Environmental Life Cycle Assessment (E-LCA). The E-LCA is based on the standards ISO 14040 and 14044. The Environmental Footprint methodology was the one used in this study to evaluate the environmental impacts. The results for the baseline cases show that the food proteins coming from whey have the highest environmental impact on ecosystems compared to the other proteins sources: 7.5 and 15.9 folds higher than soybean and gluten, respectively. Comparing feed soybean and gluten, soybean has an environmental impact on human health 195.1 folds higher. In the case of biofertilizers, synthetic struvite has higher impacts than ammonium sulfate: 15.3 (ecosystems) and 11.8 (human health) fold, respectively. The results shown in the present study will be used as a reference to demonstrate the better environmental performance of the bio-based products obtained through the VALUEWASTE solution. Other originalities that the E-LCA performed in the VALUEWASTE project provides are the diverse direct implications on investment and policies. On one hand, better environmental performance will serve to remove the barriers linked to these kinds of technologies, boosting the investment that is backed by the E-LCA. On the other hand, it will be a germ to design new policies fostering these types of solutions to achieve two of the key targets of the European Community: being self-sustainable and carbon neutral.

Keywords: anaerobic digestion, biofertilizers, circular economy, nutrients recovery

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Authors: Aberdam Edith, Sangari Linda, Petit Isabelle, Aberdam Daniel

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Background and Rationale: Transparent avascularised cornea is essential for normal vision and depends on limbal stem cells (LSC) that reside between the cornea and the conjunctiva. Ocular burns or injuries may destroy the limbus, causing limbal stem cell deficiency (LSCD). The cornea becomes vascularised by invaded conjunctival cells, the stroma is scarring, resulting in corneal opacity and loss of vision. Grafted autologous limbus or cultivated autologous LCS can restore the vision, unless the two eyes are affected. Alternative cellular sources have been tested in the last decades, including oral mucosa or hair follicle epithelial cells. However, only partial success has been achieved by the use of these cells since they were not able to uniformly commit into corneal epithelial cells. Human pluripotent stem cells (iPSC) display both unlimited growth capacity and ability to differentiate into any cell type. Our goal was to design a standardized and reproducible protocol to produce transplantable autologous LSC from patients through cell reprogramming technology. Methodology: First, keratinocyte primary culture was established from a small number of plucked hair follicles of healthy donors. The resulting epithelial cells were reprogrammed into induced pluripotent stem cells (iPSCs) and further differentiate into corneal epithelial cells (CEC), according to a robust protocol that recapitulates the main step of corneal embryonic development. qRT-PCR analysis and immunofluorescent staining during the course of differentiation confirm the expression of stage specific markers of corneal embryonic lineage. First appear ectodermal progenitor-specific cytokeratins K8/K18, followed at day 7 by limbal-specific PAX6, TP63 and cytokeratins K5/K14. At day 15, K3/K12+-corneal cells are present. To amplify the iPSC-derived LSC (named COiPSC), intact small epithelial colonies were detached and cultivated in limbal cell-specific medium. In that culture conditions, the COiPSC can be frozen and thaw at any passage, while retaining their corneal characteristics for at least eight passages. To evaluate the potential of COiPSC as an alternative ocular toxicity model, COiPSC were treated at passage P0 to P4 with increasing amounts of SDS and Benzalkonium. Cell proliferation and apoptosis of treated cells was compared to LSC and the SV40-immortalized human corneal epithelial cell line (HCE) routinely used by cosmetological industrials. Of note, HCE are more resistant to toxicity than LSC. At P0, COiPSC were systematically more resistant to chemical toxicity than LSC and even to HCE. Remarkably, this behavior changed with passage since COiPSC at P2 became identical to LSC and thus closer to physiology than HCE. Comparative transcriptome analysis confirmed that COiPSC from P2 are similar to a mixture of LSC and CEC. Finally, by organotypic reconstitution assay, we demonstrated the ability of COiPSC to produce a 3D corneal epithelium on a stromal equivalent made of keratocytes. Conclusion: COiPSC could become valuable for two main applications: (1) an alternative robust tool to perform, in a reproducible and physiological manner, toxicity assays for cosmetic products and pharmacological tests of drugs. (2). COiPSC could become an alternative autologous source for cornea transplantation for LSCD.

Keywords: Limbal stem cell deficiency, iPSC, cornea, limbal stem cells

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Authors: A. Abdi, A. Abbasi Daloee, A. Barari

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Aim: The adaptations that occur in human body after doing exercises training are a factor to help healthy people stay away from certain diseases. One of the main adaptations is a change in blood circulation, especially in vessels. The increase of capillary density is dependent on the balance between angiogenic and angiostatic factors. Most studies show that the changes made to angiogenic developmental factors resulted from physical exercises indicate the low level of stimulators compared with inhibitors. It is believed that the plasma level of VEGF-A, the important angiogenic factor, is reduced after physical exercise. Findings indicate that the extract of crataegus plant reduces the platelet-derived growth factor receptor (PDGFR) autophosphorylation in human's fibroblast. More importantly, crataegus (1 to 100 mg in liter) clearly leads to the inhibition of PDGFR autophosphorylation in vascular smooth muscle cells (VSMCs). Angiogenesis is a process that can be classified into physiological and pathophysiological forms. collagen XVIII is a part of extracellular protein and heparan sulfate proteoglycans in vascular epithelial and endothelial basement membrane cause the release of endostatin from noncollagenous collagen XVIII. Endostatin inhibits the growth of endothelial cells, inhibits angiogenesis, weakens different types of cancer, and the growth of tumors. The purpose of the current study was to investigate the effect of intensive aerobic running with or without aqueous extraction of black Crataegus elbursensis on Collagen XVIII in male rats. Design: Thirty-two Wistar male rats (4-6 weeks old, 125-135 gr weight) were acquired from the Pasteur's Institute (Amol, Mazandaran), and randomly assigned into control (n = 16) and training (n = 16) groups. Rats were further divided into saline-control (SC) (n=8), saline-training (ST) (n=8), crataegus pentaegyna extraction -control (CPEC) (n=8), and crataegus pentaegyna extraction - training (CPET) (n=8). The control (SC and CPEC) groups remained sedentary; whereas the training groups underwent a high running exercise program. plasma were excised and immediately frozen in liquid nitrogen. Statistical analysis was performed using a one way analysis of variance and Tukey test. Significance was accepted at P = 0.05. Results: The results show that aerobic exercise group had the highest concentration collagen XVIII compared to other groups and then respectively black crataegus, training-crataegus and control groups. Conclusion: In general, researchers in this study concluded that the increase of collagen XVIII (albeit insignificant) as a result of physical activity and consumption of black crataegus extract could possibly serve as a regional inhibitor of angiogenesis and another evidence for the anti-cancer effects of physical activities. Since the research has not managed in this study to measure the amount of plasma endostatin, it is suggested that both indices are measured with important angiogenic factors so that we can have a more accurate interpretation of changes to angiogenic and angiostatic factors resulted from physical exercises.

Keywords: aerobic running, Crataegus elbursensis, Collagen XVIII

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Authors: Reyhan Özdoğan, Mithat Çelebi, Özgür Ceylan, Mehmet Arif Kaya

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Super absorbent polymers (SAPs) are defined as materials that can absorb huge amount of water or aqueous solution in comparison to their own mass and retain in their lightly cross-linked structure. SAPs were produced from water soluble monomers via polymerization subsequently controlled crosslinking. SAPs are generally used for water absorbing applications such as baby diapers, patient or elder pads and other hygienic product industries. Crosslinking density (CD) of SAP structure is an essential factor for water absortion capacity (WAC). Low internal CD leads to high WAC values and vice versa. However, SAPs have low CD and high swelling capacities and tend to disintegrate when pressure is applied upon them, so SAPs under load cannot absorb liquids effectively. In order to prevent this undesired situation and to obtain suitable SAP structures having high swelling capacity and ability to work under load, surface crosslinking can be the answer. In industry, these superabsorbent gels are mostly produced via solution polymerization and then they need to be dried, grinded, sized, post polymerized and finally surface croslinked (involves spraying of a crosslinking solution onto dried and grinded SAP particles, and then curing by heat). It can easily be seen that these steps are time consuming and should be handled carefully for the desired final product. If we could synthesize desired final SAPs using less processes it will help reducing time and production costs which are very important for any industries. In this study, synthesis of SAPs were achieved successfully by inverse suspension (Pickering type) polymerization and subsequently in-situ surface cross-linking via using proper surfactants in high boiling point solvents. Our one-pot synthesis of surface cross-linked SAPs invovles only one-step for preparation, thus it can be said that this technique exhibits more preferable characteristic for the industry in comparison to conventional methods due to its one-step easy process. Effects of different surface crosslinking agents onto properties of poly(acrylic acid-co-sodium acrylate) based SAPs are investigated. Surface crosslink degrees are evaluated by swelling under load (SUL) test. It was determined water absorption capacities of obtained SAPs decrease with the increasing surface crosslink density while their mechanic properties are improved.

Keywords: inverse suspension polymerization, polyacrylic acid, super absorbent polymers (SAPs), surface crosslinking, sodium polyacrylate

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Authors: Adriana Garcia G., Alberto A. Escobar P., Amira D. Calvo L., Gabriel Lizama U., Alejandro Zepeda P., Fernando Martínez B., Susana Rincón A.

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Different studies have recently been focused on the use of antioxidants such as polyphenols because of to its anticarcinogenic capacity. However, these compounds are highly sensible to environmental factors such as light and heat, so lose its long-term stability, besides possess an astringent and bitter taste. Nevertheless, the polyphenols can be protected by microcapsule formulation. In this sense, a rich source of polyphenols is sorghum, besides presenting a high starch content. Due to the above, the aim of this work was to obtain modified starches from sorghum by extrusion to encapsulate polyphenols the sorghum by spray drying. Polyphenols were extracted by ethanol solution from sorghum (Pajarero/red) and determined by the method of Folin-Ciocalteu, obtaining GAE at 30 mg/g. Moreover, was extracted starch of sorghum (Sinaloense/white) through wet milling (yield 32 %). The hydrolyzed starch was modified with three treatments: acetic anhydride (2.5g/100g), sodium tripolyphosphate (4g/100g), and sodium tripolyphosphate/ acetic anhydride (2g/1.25g by each 100 g) by extrusion. Processing conditions of extrusion were as follows: barrel temperatures were of 60, 130 and 170 °C at the feeding, transition, and high-pressure extrusion zones, respectively. Analysis of Fourier Transform Infrared spectroscopy (FTIR), showed bands exhibited of acetyl groups (1735 cm-1) and phosphates (1170 cm-1, 910 cm-1 and 525 cm-1), indicating the respective modification of starch. Besides, all modified starches not developed viscosity, which is a characteristic required for use in the encapsulation of polyphenols using the spray drying technique. As result of the modification starch, was obtained a water solubility index (WSI) from 33.8 to 44.8 %, and crystallinity from 8 to 11 %, indicating the destruction of the starch granule. Afterwards, microencapsulation of polyphenols was developed by spray drying, with a blend of 10 g of modified starch, 60 ml polyphenol extract and 30 ml of distilled water. Drying conditions were as follows: inlet air temperature 150 °C ± 1, outlet air temperature 80°C ± 5. As result of the microencapsulation: were obtained yields of 56.8 to 77.4 % and an efficiency of encapsulation from 84.6 to 91.4 %. The FTIR analysis showed evidence of microcapsules loaded with polyphenols in bands 1042 cm-1, 1038 cm-1 and 1148 cm-1. Analysis Differential scanning calorimetry (DSC) showed transition temperatures from 144.1 to 173.9 °C. For the order hand, analysis of Scanning Electron Microscopy (SEM), were observed rounded surfaces with concavities, typical feature of microcapsules produced by spray drying, how result of rapid evaporation of water. Finally, the modified starches were obtained by extrusion with good characteristics for use as cover materials by spray drying, where the phosphorylated starch was the best treatment in this work, according to the encapsulation yield, efficiency, and transition temperature.

Keywords: encapsulation, extrusion, modified starch, polyphenols, spray drying

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Authors: Wedad Albalawi

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The mathematical inequalities have been the core of mathematical study and used in almost all branches of mathematics as well in various areas of science and engineering. The inequalities by Hardy, Littlewood and Polya were the first significant composition of several science. This work presents fundamental ideas, results and techniques, and it has had much influence on research in various branches of analysis. Since 1934, various inequalities have been produced and studied in the literature. Furthermore, some inequalities have been formulated by some operators; in 1989, weighted Hardy inequalities have been obtained for integration operators. Then, they obtained weighted estimates for Steklov operators that were used in the solution of the Cauchy problem for the wave equation. They were improved upon in 2011 to include the boundedness of integral operators from the weighted Sobolev space to the weighted Lebesgue space. Some inequalities have been demonstrated and improved using the Hardy–Steklov operator. Recently, a lot of integral inequalities have been improved by differential operators. Hardy inequality has been one of the tools that is used to consider integrity solutions of differential equations. Then, dynamic inequalities of Hardy and Coposon have been extended and improved by various integral operators. These inequalities would be interesting to apply in different fields of mathematics (functional spaces, partial differential equations, mathematical modeling). Some inequalities have been appeared involving Copson and Hardy inequalities on time scales to obtain new special version of them. A time scale is an arbitrary nonempty closed subset of the real numbers. Then, the dynamic inequalities on time scales have received a lot of attention in the literature and has become a major field in pure and applied mathematics. There are many applications of dynamic equations on time scales to quantum mechanics, electrical engineering, neural networks, heat transfer, combinatorics, and population dynamics. This study focuses on Hardy and Coposon inequalities, using Steklov operator on time scale in double integrals to obtain special cases of time-scale inequalities of Hardy and Copson on high dimensions. The advantage of this study is that it uses the one-dimensional classical Hardy inequality to obtain higher dimensional on time scale versions that will be applied in the solution of the Cauchy problem for the wave equation. In addition, the obtained inequalities have various applications involving discontinuous domains such as bug populations, phytoremediation of metals, wound healing, maximization problems. The proof can be done by introducing restriction on the operator in several cases. The concepts in time scale version such as time scales calculus will be used that allows to unify and extend many problems from the theories of differential and of difference equations. In addition, using chain rule, and some properties of multiple integrals on time scales, some theorems of Fubini and the inequality of H¨older.

Keywords: time scales, inequality of hardy, inequality of coposon, steklov operator

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Authors: Rauan Sakenov, Peter Bukovics, Peter Gaszler, Veronika Tokacs-Kollar, Beata Bugyi

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FH2 (formin homology-2) domains of several proteins, collectively known as formins, including DAAM, DAAM1 and mDia1, promote G-actin nucleation and elongation. FH2 domains of these formins exist as oligomers. Chain dimerization by ring structure formation serves as a structural basis for actin polymerization function of FH2 domain. Proper single chain configuration and specific interactions between its various regions are necessary for individual chains to form a dimer functional in G-actin nucleation and elongation. FH1 and WH2 domain-containing formins were shown to behave as intrinsically disordered proteins. Thus, the aim of this research was to study structural dynamics of FH2 domain of DAAM. To investigate structural features of FH2 domain of DAAM, molecular dynamics simulation of chain A of FH2 domain of DAAM solvated in water box in 50 mM NaCl was conducted at temperatures from 293.15 to 353.15K, with VMD 1.9.2, NAMD 2.14 and Amber Tools 21 using 2z6e and 1v9d PDB structures of DAAM was obtained on I-TASSER webserver. Calcium and ATP bound G-actin 3hbt PDB structure was used as a reference protein with well-described structural dynamics of denaturation. Topology and parameter information of CHARMM 2012 additive all-atom force fields for proteins, carbohydrate derivatives, water and ions were used in NAMD 2.14 and ff19SB force field for proteins in Amber Tools 21. The systems were energy minimized for the first 1000 steps, equilibrated and produced in NPT ensemble for 1ns using stochastic Langevin dynamics and the particle mesh Ewald method. Our root-mean square deviation (RMSD) analysis of molecular dynamics of chain A of FH2 domains of DAAM revealed similar insignificant changes of total molecular average RMSD values of FH2 domain of these formins at temperatures from 293.15 to 353.15K. In contrast, total molecular average RMSD values of G-actin showed considerable increase at 328K, which corresponds to the denaturation of G-actin molecule at this temperature and its transition from native, ordered, to denatured, disordered, state which is well-described in the literature. RMSD values of lasso and tail regions of chain A of FH2 domain of DAAM exhibited higher than total molecular average RMSD at temperatures from 293.15 to 353.15K. These regions are functional in intra- and interchain interactions and contain highly conserved tryptophan residues of lasso region, highly conserved GNYMN sequence of post region and amino acids of the shell of hydrophobic pocket of the salt bridge between Arg171 and Asp321, which are important for structural stability and ordered state of FH2 domain of DAAM and its functions in FH2 domain dimerization. In conclusion, higher than total molecular average RMSD values of lasso and post regions of chain A of FH2 domain of DAAM may explain disordered state of FH2 domain of DAAM at temperatures from 293.15 to 353.15K. Finally, absence of marked transition, in terms of significant changes in average molecular RMSD values between native and denatured states of FH2 domain of DAAM at temperatures from 293.15 to 353.15K, can make it possible to attribute these formins to the group of intrinsically disordered proteins rather than to the group of intrinsically ordered proteins such as G-actin.

Keywords: FH2 domain, DAAM, formins, molecular modelling, computational biophysics

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Authors: Saule Balmagambetova, Zhenisgul Tlegenova, Saule Madinova

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Cardiotoxicity associated with anticancer treatment, now defined as cancer therapy-related cardiac dysfunction (CTRCD), accompanies cancer patients and negatively impacts their survivorship. Currently, a cardio-oncological service is being created in Kazakhstan based on the provisions of the European Society of Cardio-oncology (ESC) Guidelines. In the frames of a pilot project, a cohort study on CTRCD conditions was initiated at the Aktobe Cancer center. One hundred twenty-eight newly diagnosed breast cancer patients started on doxorubicin and/or trastuzumab were recruited. Echocardiography with global longitudinal strain (GLS) assessment, biomarkers panel (cardiac troponin (cTnI), brain natriuretic peptide (BNP), myeloperoxidase (MPO), galectin-3 (Gal-3), D-dimers, C-reactive protein (CRP)), and other tests were performed at baseline and every three months. Patients were stratified by the cardiovascular risks according to the ESC recommendations and allocated into the risk groups during the pre-treatment visit. Of them, 10 (7.8%) patients were assigned to the high-risk group, 48 (37.5%) to the medium-risk group, and 70 (54.7%) to the low-risk group, respectively. High-risk patients have been receiving their cardioprotective treatment from the outset. Patients were also divided by treatment - in the anthracycline-based 83 (64.8%), in trastuzumab- only 13 (10.2%), and in the mixed anthracycline/trastuzumab group 32 individuals (25%), respectively. Mild symptomatic CTRCD was revealed and treated in 2 (1.6%) participants, and a mild asymptomatic variant in 26 (20.5%). Mild asymptomatic conditions are defined as left ventricular ejection fraction (LVEF) ≥50% and further relative reduction in GLS by >15% from baseline and/or a further rise in cardiac biomarkers. The listed biomarkers were assessed longitudinally in repeated-measures linear regression models during 12 months of observation. The associations between changes in biomarkers and CTRCD and between changes in biomarkers and LVEF were evaluated. Analysis by risk groups revealed statistically significant differences in baseline LVEF scores (p 0.001), BNP (p 0.0075), and Gal-3 (p 0.0073). Treatment groups found no statistically significant differences at baseline. After 12 months of follow-up, only LVEF values showed a statistically significant difference by risk groups (p 0.0011). When assessing the temporal changes in the studied parameters for all treatment groups, there were statistically significant changes from visit to visit for LVEF (p 0.003); GLS (p 0.0001); BNP (p<0.00001); MPO (p<0.0001); and Gal-3 (p<0.0001). No moderate or strong correlations were found between the biomarkers values and LVEF, between biomarkers and GLS. Between the biomarkers themselves, a moderate, close to strong correlation was established between cTnI and D-dimer (r 0.65, p<0.05). The dose-dependent effect of anthracyclines has been confirmed: the summary dose has a moderate negative impact on GLS values: -r 0.31 for all treatment groups (p<0.05). The present study found myeloperoxidase as a promising biomarker of cardiac dysfunction in the mixed anthracycline/trastuzumab treatment group. The hazard of CTRCD increased by 24% (HR 1.21; 95% CI 1.01;1.73) per doubling in baseline MPO value (p 0.041). Increases in BNP were also associated with CTRCD (HR per doubling, 1.22; 95% CI 1.12;1.69). No cases of chemotherapy discontinuation due to cardiotoxic complications have been recorded. Further observations are needed to gain insight into the ability of biomarkers to predict CTRCD onset.

Keywords: breast cancer, chemotherapy, cardiotoxicity, Kazakhstan

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Authors: Ning Chia

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Each of the three different language texts (Manchu, Russian, and Latin) of the 1689 Treaty of Nerchinsk bore official seals from Imperial Russia and Qing China. These seals have received no academic attention, yet they can reveal a site of a layered and shared material, cultural, political, and diplomatic world of the time in Eastern Eurasia. The very different seal selections from both empires while ratifying the Treaty of Beijing in 1860 have obtained no scholarly advertency either; they can also explicate a tremendously changed relationship with visual and material manifestation. Exploring primary sources in Manchu, Russian, and Chinese languages as well as the images of the visual seals, this study investigates the reasons and purposes of utilizing official seals for the treaty agreement. A refreshed understanding of Russian-Qing diplomacy will be developed by pursuing the following aspects: (i) Analyzing the iconographic meanings of each seal insignia and unearthing a competitive, yet symbols-delivered and seal-generated, 'dialogue' between the two empires (ii) Contextualizing treaty seals within the historical seal cultures, and discovering how domestic seal system in each empire’s political institution developed into treaty-defined bilateral relations (iii) Expounding the seal confiding in each empire’s daily governing routines, and annotating the trust in the seal as a quested promise from the opponent negotiator to fulfill the treaty terms (iv) Contrasting the two seal traditions along two civilization-lines, Eastern vs. Western, and dissecting how the two styles of seal emblems affected the cross-cultural understanding or misunderstanding between the two empires (v) Comprehending the history-making events from the substantial resources such as the treaty seals, and grasping why the seals for the two treaties, so different in both visual design and symbolic value, were chosen in the two relationship eras (vi) Correlating the materialized seal 'expression' and the imperial worldviews based on each empire’s national/or power identity, and probing the seal-represented 'rule under the Heaven' assumption of China and Russian rising role in 'European-American imperialism … centered on East Asia' (Victor Shmagin, 2020). In conclusion, the impact of official seals on diplomatic treaties needs profound knowledge in seal history, insignia culture, and emblem belief to be able to comprehend. The official seals in both Imperial Russia and Qing China belonged to a particular statecraft art in a specific material and visual form. Once utilized in diplomatic treaties, the meticulously decorated and politically institutionalized seals were transformed from the determinant means for domestic administration and social control into the markers of an empire’s sovereign authority. Overlooked in historical practice, the insignia seal created a wire of 'visual contest' between the two rival powers. Through this material lens, the scholarly knowledge of the Russian-Qing diplomatic relationship will be significantly upgraded. Connecting Russian studies, Qing/Chinese studies, and Eurasian studies, this study also ties material culture, political culture, and diplomatic culture together. It promotes the study of official seals and emblem symbols in worldwide diplomatic history.

Keywords: Russia-Qing diplomatic relation, Treaty of Beijing (1860), Treaty of Nerchinsk (1689), Treaty seals

Procedia PDF Downloads 193

Authors: Maddalena Arigoni, Maria Luisa Ratto, Raffaele A. Calogero, Luca Alessandri

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The presence of tumor heterogeneity, where distinct cancer cells exhibit diverse morphological and phenotypic profiles, including gene expression, metabolism, and proliferation, poses challenges for molecular prognostic markers and patient classification for targeted therapies. Understanding the causes and progression of cancer requires research efforts aimed at characterizing heterogeneity, which can be facilitated by evolving single-cell sequencing technologies. However, analyzing single-cell data necessitates computational methods that often lack objective validation. Therefore, the establishment of benchmarking datasets is necessary to provide a controlled environment for validating bioinformatics tools in the field of single-cell oncology. Benchmarking bioinformatics tools for single-cell experiments can be costly due to the high expense involved. Therefore, datasets used for benchmarking are typically sourced from publicly available experiments, which often lack a comprehensive cell annotation. This limitation can affect the accuracy and effectiveness of such experiments as benchmarking tools. To address this issue, we introduce omics benchmark experiments designed to evaluate bioinformatics tools to depict the heterogeneity in single-cell tumor experiments. We conducted single-cell RNA sequencing on six lung cancer tumor cell lines that display resistant clones upon treatment of EGFR mutated tumors and are characterized by driver genes, namely ROS1, ALK, HER2, MET, KRAS, and BRAF. These driver genes are associated with downstream networks controlled by EGFR mutations, such as JAK-STAT, PI3K-AKT-mTOR, and MEK-ERK. The experiment also featured an EGFR-mutated cell line. Using 10XGenomics platform with cellplex technology, we analyzed the seven cell lines together with a pseudo-immunological microenvironment consisting of PBMC cells labeled with the Biolegend TotalSeq™-B Human Universal Cocktail (CITEseq). This technology allowed for independent labeling of each cell line and single-cell analysis of the pooled seven cell lines and the pseudo-microenvironment. The data generated from the aforementioned experiments are available as part of an online tool, which allows users to define cell heterogeneity and generates count tables as an output. The tool provides the cell line derivation for each cell and cell annotations for the pseudo-microenvironment based on CITEseq data by an experienced immunologist. Additionally, we created a range of pseudo-tumor tissues using different ratios of the aforementioned cells embedded in matrigel. These tissues were analyzed using 10XGenomics (FFPE samples) and Curio Bioscience (fresh frozen samples) platforms for spatial transcriptomics, further expanding the scope of our benchmark experiments. The benchmark experiments we conducted provide a unique opportunity to evaluate the performance of bioinformatics tools for detecting and characterizing tumor heterogeneity at the single-cell level. Overall, our experiments provide a controlled and standardized environment for assessing the accuracy and robustness of bioinformatics tools for studying tumor heterogeneity at the single-cell level, which can ultimately lead to more precise and effective cancer diagnosis and treatment.

Keywords: single cell omics, benchmark, spatial transcriptomics, CITEseq

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Authors: Mathieu Bourdeau, Philippe Basset, Julien Waeytens, Elyes Nefzaoui

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As residential buildings account for a third of the overall energy consumption and greenhouse gas emissions in Europe, building energy modeling is an essential tool to reach energy efficiency goals. In the energy modeling process, calibration is a mandatory step to obtain accurate and reliable energy simulations. Nevertheless, the comparison between simulation results and the actual building energy behavior often highlights a significant performance gap. The literature discusses different origins of energy performance gaps, from building design to building operation. Then, building operation description in energy models, especially energy usages and users’ behavior, plays an important role in the reliability of simulations but is also the most accessible target for post-occupancy energy management and optimization. Therefore, the present study aims to discuss results on the calibration ofresidential building energy models using real operation data. Data are collected through a sensor network of more than 180 sensors and advanced energy meters deployed in three collective residential buildings undergoing major retrofit actions. The sensor network is implemented at building scale and in an eight-apartment sample. Data are collected for over one year and half and coverbuilding energy behavior – thermal and electricity, indoor environment, inhabitants’ comfort, occupancy, occupants behavior and energy uses, and local weather. Building energy simulations are performed using a physics-based building energy modeling software (Pleaides software), where the buildings’features are implemented according to the buildingsthermal regulation code compliance study and the retrofit project technical files. Sensitivity analyses are performed to highlight the most energy-driving building features regarding each end-use. These features are then compared with the collected post-occupancy data. Energy-driving features are progressively replaced with field data for a step-by-step calibration of the energy model. Results of this study provide an analysis of energy performance gap on an existing residential case study under deep retrofit actions. It highlights the impact of the different building features on the energy behavior and the performance gap in this context, such as temperature setpoints, indoor occupancy, the building envelopeproperties but also domestic hot water usage or heat gains from electric appliances. The benefits of inputting field data from an extensive instrumentation campaign instead of standardized scenarios are also described. Finally, the exhaustive instrumentation solution provides useful insights on the needs, advantages, and shortcomings of the implemented sensor network for its replicability on a larger scale and for different use cases.

Keywords: calibration, building energy modeling, performance gap, sensor network

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Authors: Maryam Hajrezaie, Mahmood Ameen Abdulla, Nazia Abdul Majid, Hapipa Mohd Ali, Pouya Hassandarvish, Maryam Zahedi Fard

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Background: Colon cancer is one of the most prevalent cancers in the world and is the third leading cause of death among cancers in both males and females. The incidence of colon cancer is ranked fourth among all cancers but varies in different parts of the world. Cancer chemoprevention is defined as the use of natural or synthetic compounds capable of inducing biological mechanisms necessary to preserve genomic fidelity. Andrographolide is the major labdane diterpenoidal constituent of the plant Andrographis paniculata (family Acanthaceae), used extensively in the traditional medicine. Extracts of the plant and their constituents are reported to exhibit a wide spectrum of biological activities of therapeutic importance. Laboratory animal model studies have provided evidence that Andrographolide play a role in inhibiting the risk of certain cancers. Objective: Our aim was to evaluate the chemopreventive efficacy of the Andrographolide in the AOM induced rat model. Methods: To evaluate inhibitory properties of andrographolide on colonic aberrant crypt foci (ACF), five groups of 7-week-old male rats were used. Group 1 (control group) were fed with 10% Tween 20 once a day, Group 2 (cancer control) rats were intra-peritoneally injected with 15 mg/kg Azoxymethan, Gropu 3 (drug control) rats were injected with 15 mg/kg azoxymethan and 5-Flourouracil, Group 4 and 5 (experimental groups) were fed with 10 and 20 mg/kg andrographolide each once a day. After 1 week, the treatment group rats received subcutaneous injections of azoxymethane, 15 mg/kg body weight, once weekly for 2 weeks. Control rats were continued on Tween 20 feeding once a day and experimental groups 10 and 20 mg/kg andrographolide feeding once a day for 8 weeks. All rats were sacrificed 8 weeks after the azoxymethane treatment. Colons were evaluated grossly and histopathologically for ACF. Results: Administration of 10 mg/kg and 20 mg/kg andrographolide were found to be effectively chemoprotective, as evidenced microscopily and biochemically. Andrographolide suppressed total colonic ACF formation up to 40% to 60%, respectively, when compared with control group. Pre-treatment with andrographolide, significantly reduced the impact of AOM toxicity on plasma protein and urea levels as well as on plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and gamma-glutamyl transpeptidase (GGT) activities. Grossly, colorectal specimens revealed that andrographolide treatments decreased the mean score of number of crypts in AOM-treated rats. Importantly, rats fed andrographolide showed 75% inhibition of foci containing four or more aberrant crypts. The results also showed a significant increase in glutathione (GSH), superoxide dismutase (SOD), nitric oxide (NO), and Prostaglandin E2 (PGE2) activities and a decrease in malondialdehyde (MDA) level. Histologically all treatment groups showed a significant decrease of dysplasia as compared to control group. Immunohistochemical staining showed up-regulation of Hsp70 and down-regulation of Bax proteins. Conclusion: The current study demonstrated that Andrographolide reduce the number of ACF. According to these data, Andrographolide might be a promising chemoprotective activity, in a model of AOM-induced in ACF.

Keywords: chemopreventive, andrographolide, colon cancer, aberrant crypt foci (ACF)

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Authors: John Christian Lequiron, Gerry Bagtasa, Olivia Cabrera, Leoncio Amadore, Tolentino Moya

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Changes in air surface temperature play an important role in the Philippine’s economy, industry, health, and food production. While increasing global mean temperature in the recent several decades has prompted a number of climate change and variability studies in the Philippines, most studies still focus on rainfall and tropical cyclones. This study aims to investigate the trend and variability of observed air surface temperature and determine its major influencing factor/s in the Philippines. A non-parametric Mann-Kendall trend test was applied to monthly mean temperature of 17 synoptic stations covering 56 years from 1960 to 2015 and a mean change of 0.58 °C or a positive trend of 0.0105 °C/year (p < 0.05) was found. In addition, wavelet decomposition was used to determine the frequency of temperature variability show a 12-month, 30-80-month and more than 120-month cycles. This indicates strong annual variations, interannual variations that coincide with ENSO events, and interdecadal variations that are attributed to PDO and CO2 concentrations. Air surface temperature was also correlated with smoothed sunspot number and galactic cosmic rays, the results show a low to no effect. The influence of ENSO teleconnection on temperature, wind pattern, cloud cover, and outgoing longwave radiation on different ENSO phases had significant effects on regional temperature variability. Particularly, an anomalous anticyclonic (cyclonic) flow east of the Philippines during the peak and decay phase of El Niño (La Niña) events leads to the advection of warm southeasterly (cold northeasterly) air mass over the country. Furthermore, an apparent increasing cloud cover trend is observed over the West Philippine Sea including portions of the Philippines, and this is believed to lessen the effect of the increasing air surface temperature. However, relative humidity was also found to be increasing especially on the central part of the country, which results in a high positive trend of heat index, exacerbating the effects on human discomfort. Finally, an assessment of gridded temperature datasets was done to look at the viability of using three high-resolution datasets in future climate analysis and model calibration and verification. Several error statistics (i.e. Pearson correlation, Bias, MAE, and RMSE) were used for this validation. Results show that gridded temperature datasets generally follows the observed surface temperature change and anomalies. In addition, it is more representative of regional temperature rather than a substitute to station-observed air temperature.

Keywords: air surface temperature, carbon dioxide, ENSO, galactic cosmic rays, smoothed sunspot number

Procedia PDF Downloads 295

Authors: Xianghe Dai, Dennis Lam, Therese Sheehan, Naveed Rehman, Jie Yang

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Concrete is one of the most used construction materials that may be made onsite as fresh concrete and then placed in formwork to produce the desired shapes of structures. It has been recognized that the raw materials and mix proportion of concrete dominate the mechanical characteristics of hardened concrete, and the curing method and environment applied to the concrete in early stages of hardening will significantly influence the concrete properties, such as compressive strength, durability, permeability etc. In construction practice, there are various curing methods to maintain the presence of mixing water throughout the early stages of concrete hardening. They are also beneficial to concrete in hot weather conditions as they provide cooling and prevent the evaporation of water. Such methods include ponding or immersion, spraying or fogging, saturated wet covering etc. Also there are various curing methods that may be implemented to decrease the level of water lost which belongs to the concrete surface, such as putting a layer of impervious paper, plastic sheeting or membrane on the concrete to cover it. In the concrete material laboratory, accelerated strength gain methods supply the concrete with heat and additional moisture by applying live steam, coils that are subject to heating or pads that have been warmed electrically. Currently when determining the mechanical parameters of a concrete, the concrete is usually sampled from fresh concrete on site and then cured and tested in laboratories where standardized curing procedures are adopted. However, in engineering practice, curing procedures in the construction sites after the placing of concrete might be very different from the laboratory criteria, and this includes some standard curing procedures adopted in the laboratory that can’t be applied on site. Sometimes the contractor compromises the curing methods in order to reduce construction costs etc. Obviously the difference between curing procedures adopted in the laboratory and those used on construction sites might over- or under-estimate the real concrete quality. This paper presents the effect of three typical curing methods (air curing, water immersion curing, plastic film curing) and of maintaining concrete in steel moulds on the compressive strength development of normal concrete. In this study, Portland cement with 30% fly ash was used and different curing periods, 7 days, 28 days and 60 days were applied. It was found that the highest compressive strength was observed from concrete samples to which 7-day water immersion curing was applied and from samples maintained in steel moulds up to the testing date. The research results implied that concrete used as infill in steel tubular members might develop a higher strength than predicted by design assumptions based on air curing methods. Wrapping concrete with plastic film as a curing method might delay the concrete strength development in the early stages. Water immersion curing for 7 days might significantly increase the concrete compressive strength.

Keywords: compressive strength, air curing, water immersion curing, plastic film curing, maintaining in steel mould, comparison

Procedia PDF Downloads 278

Authors: Deepak Loura

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Production and productivity of several crops in the country continue to be adversely affected by biotic (e.g., Insect-pests and diseases) and abiotic (e.g., water temperature and salinity) stresses. Over-dependence on pesticides and other chemicals is economically non-viable for the resource-poor farmers of our country. Further, pesticides can potentially affect human and environmental safety. While traditional breeding techniques and proper- management strategies continue to play a vital role in crop improvement, we need to judiciously use biotechnology approaches for the development of genetically modified crops addressing critical problems in the improvement of crop plants for sustainable agriculture. Modern biotechnology can help to increase crop production, reduce farming costs, and improve food quality and the safety of the environment. Genetic engineering is a new technology which allows plant breeders to produce plants with new gene combinations by genetic transformation of crop plants for improvement of agronomic traits. Advances in recombinant DNA technology have made it possible to have genes between widely divergent species to develop genetically modified or genetically engineered plants. Plant genetic engineering provides the strength to harness useful genes and alleles from indigenous microorganisms to enrich the gene pool for developing genetically modified (GM) crops that will have inbuilt (inherent) resistance to insect pests, diseases, and abiotic stresses. Plant biotechnology has made significant contributions in the past 20 years in the development of genetically engineered or genetically modified crops with multiple benefits. A variety of traits have been introduced in genetically engineered crops which include (i) herbicide resistance. (ii) pest resistance, (iii) viral resistance, (iv) slow ripening of fruits and vegetables, (v) fungal and bacterial resistance, (vi) abiotic stress tolerance (drought, salinity, temperature, flooding, etc.). (vii) quality improvement (starch, protein, and oil), (viii) value addition (vitamins, micro, and macro elements), (ix) pharmaceutical and therapeutic proteins, and (x) edible vaccines, etc. Multiple genes in transgenic crops can be useful in developing durable disease resistance and a broad insect-control spectrum and could lead to potential cost-saving advantages for farmers. The development of transgenic to produce high-value pharmaceuticals and the edible vaccine is also under progress, which requires much more research and development work before commercially viable products will be available. In addition, molecular-aided selection (MAS) is now routinely used to enhance the speed and precision of plant breeding. Newer technologies need to be developed and deployed for enhancing and sustaining agricultural productivity. There is a need to optimize the use of biotechnology in conjunction with conventional technologies to achieve higher productivity with fewer resources. Therefore, genetic modification/ engineering of crop plants assumes greater importance, which demands the development and adoption of newer technology for the genetic improvement of crops for increasing crop productivity.

Keywords: biotechnology, plant genetic engineering, genetically modified, biotic, abiotic, disease resistance

Procedia PDF Downloads 56

Authors: Alon Cohen, Jeffrey Kantor, Shalom Levy

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The use of the audit risk model in auditing has faced limitations and difficulties, leading auditors to rely on a conceptual level of its application. The qualitative approach to assessing risks has resulted in different risk assessments, affecting the quality of audits and decision-making on the adoption of CAATTs. This study aims to investigate risk factors impacting the implementation of the audit risk model and propose a complementary risk-based instrument (KRIs) to form substance risk judgments and mitigate against heightened risk of material misstatement (RMM). The study addresses the question of how risk factors impact the implementation of the audit risk model, improve risk judgments, and aid in the adoption of CAATTs. The study uses a three-stage scale development procedure involving a pretest and subsequent study with two independent samples. The pretest involves an exploratory factor analysis, while the subsequent study employs confirmatory factor analysis for construct validation. Additionally, the authors test the ability of the KRIs to predict audit efforts needed to mitigate against heightened RMM. Data was collected through two independent samples involving 767 participants. The collected data was analyzed using exploratory factor analysis and confirmatory factor analysis to assess scale validity and construct validation. The suggested KRIs, comprising two risk components and seventeen risk items, are found to have high predictive power in determining audit efforts needed to reduce RMM. The study validates the suggested KRIs as an effective instrument for risk assessment and decision-making on the adoption of CAATTs. This study contributes to the existing literature by implementing a holistic approach to risk assessment and providing a quantitative expression of assessed risks. It bridges the gap between intuitive risk evaluation and the theoretical domain, clarifying the mechanism of risk assessments. It also helps improve the uniformity and quality of risk assessments, aiding audit standard-setters in issuing updated guidelines on CAATT adoption. A few limitations and recommendations for future research should be mentioned. First, the process of developing the scale was conducted in the Israeli auditing market, which follows the International Standards on Auditing (ISAs). Although ISAs are adopted in European countries, for greater generalization, future studies could focus on other countries that adopt additional or local auditing standards. Second, this study revealed risk factors that have a material impact on the assessed risk. However, there could be additional risk factors that influence the assessment of the RMM. Therefore, future research could investigate other risk segments, such as operational and financial risks, to bring a broader generalizability to our results. Third, although the sample size in this study fits acceptable scale development procedures and enables drawing conclusions from the body of research, future research may develop standardized measures based on larger samples to reduce the generation of equivocal results and suggest an extended risk model.

Keywords: audit risk model, audit efforts, CAATTs adoption, key risk indicators, sustainability

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Authors: Isaura Bonneux

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District heating and cooling (DHC) is increasingly recognized as a viable path towards sustainable heating and cooling. While some countries like Sweden and Denmark have a longstanding tradition of DHC, Belgium is lacking behind. The Northern part of Belgium, Flanders, had only a total of 95 heating networks in July 2023. Nevertheless, it is increasingly exploring its possibilities to enhance the scope of DHC. DHC is a complex energy system, requiring a lot of collaboration between various stakeholders on various levels. Therefore, it is of interest to look closer at policy-related documents at the Flemish (regional) level, as these policies set the scene for DHC development in the Flemish region. This kind of analysis has not been undertaken so far. This paper has the following research question: “Who talks about DHC, and in which way and context is DHC discussed in Flemish policy-related documents?” To answer this question, the Overton policy database was used to search and retrieve relevant policy-related documents. Overton retrieves data from governments, think thanks, NGOs, and IGOs. In total, out of the 244 original results, 117 documents between 2009 and 2023 were analyzed. Every selected document included theme keywords, policymaking department(s), date, and document type. These elements were used for quantitative data description and visualization. Further, qualitative content analysis revealed patterns and main themes regarding DHC in Flanders. Four main conclusions can be drawn: First, it is obvious from the timeframe that DHC is a new topic in Flanders with still limited attention; 2014, 2016 and 2017 were the years with the most documents, yet this number is still only 12 documents. In addition, many documents talked about DHC but not much in depth and painted it as a future scenario with a lot of uncertainty around it. The largest part of the issuing government departments had a link to either energy or climate (e.g. Flemish Environmental Agency) or policy (e.g. Socio-Economic Council of Flanders) Second, DHC is mentioned most within an ‘Environment and Sustainability’ context, followed by ‘General Policy and Regulation’. This is intuitive, as DHC is perceived as a sustainable heating and cooling technique and this analysis compromises policy-related documents. Third, Flanders seems mostly interested in using waste or residual heat as a heating source for DHC. The harbors and waste incineration plants are identified as potential and promising supply sources. This approach tries to conciliate environmental and economic incentives. Last, local councils get assigned a central role and the initiative is mostly taken by them. The policy documents and policy advices demonstrate that Flanders opts for a bottom-up organization. As DHC is very dependent on local conditions, this seems a logic step. Nevertheless, this can impede smaller councils to create DHC networks and slow down systematic and fast implementation of DHC throughout Flanders.

Keywords: district heating and cooling, flanders, overton database, policy analysis

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Authors: Elena Sherstoboeva, Elena Karzanova

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This paper explores the impact of the annexation of Crimea on the regulation of live performances and tour management of Russian pop music performers in Ukraine and of Ukrainian performers in Russia. Without a doubt, the cultural relationship between Russia and Ukraine is not limited to this issue. Yet concert markets tend to respond particularly rapidly to political, economic, and social changes, especially in Russia and Ukraine, where the high level of digital piracy means that the music businesses mainly depend upon income from performances rather than from digital rights sales. This paper argues that the rules formed in both countries after Russia’s annexation of Crimea in 2014 have contributed to the separation of a single cultural space that had existed in Soviet and Post-Soviet Russia and Ukraine before the annexation. These rules have also facilitated performers’ self-censorship and increased the politicisation of the music businesses in the two neighbouring countries. This study applies a comparative socio-legal approach to study Russian and Ukrainian live events and tour regulation. A qualitative analysis of Russian and Ukrainian national and intergovernmental legal frameworks is applied to examine formal regulations. Soviet and early post-Soviet laws and policies are also studied, but only to the extent that they help to track the changes in the Russian–Ukrainian cultural relationship. To identify and analyse the current informal rules, the study design includes in-depth semi-structured interviews with 30 live event or tour managers working in Russia and Ukraine. A case study is used to examine how the Eurovision Song Contest, an annual international competition, has played out within the Russian–Ukrainian conflict. The study suggests that modern Russian and Ukrainian frameworks for live events and tours have developed Soviet regulatory traditions when cultural policies served as a means of ideological control. At the same time, contemporary regulations mark a considerable perspective shift, as the previous rules have been aimed at maintaining close cultural connections between the Russian and Ukrainian nations. Instead of collaboration, their current frameworks mostly serve as forms of repression, implying that performers must choose only one national market in which to work. The regulatory instruments vary and often impose limitations that typically exist in non-democratic regimes to restrict foreign journalism, such as visa barriers or bans on entry. The more unexpected finding is that, in comparison with Russian law, Ukrainian regulations have created more obstacles to the organisation of live tours and performances by Russian artists in Ukraine. Yet this stems from commercial rather than political factors. This study predicts that the more economic challenges the Russian or Ukrainian music businesses face, the harsher the regulations will be regarding the organisation of live events or tours in the other country. This study recommends that international human rights organisations and non-governmental organisations develop and promote specific standards for artistic rights and freedoms, given the negative effects of the increasing politicisation of the entertainment business and cultural spheres to freedom of expression and cultural rights and pluralism.

Keywords: annexation of Crimea, artistic freedom, censorship, cultural policy

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Authors: Gabrielle Peck, Ryan Hayes

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Current large scale fire tests focus on flammability and heat release measurements. Smoke toxicity isn’t considered despite it being a leading cause of death and injury in unwanted fires. A key reason could be that the practical difficulties associated with quantifying individual toxic components present in a fire effluent often require specialist equipment and expertise. Fire effluent contains a mixture of unreactive and reactive gases, water, organic vapours and particulate matter, which interact with each other. This interferes with the operation of the analytical instrumentation and must be removed without changing the concentration of the target analyte. To mitigate the need for expensive equipment and time-consuming analysis, a portable gas analysis system was designed, constructed and tested for use in large-scale fire tests as a simpler and more robust alternative to online FTIR measurements. The novel equipment aimed to be easily portable and able to run on battery or mains electricity; be able to be calibrated at the test site; be capable of quantifying CO, CO2, O2, HCN, HBr, HCl, NOx and SO2 accurately and reliably; be capable of independent data logging; be capable of automated switchover of 7 bubblers; be able to withstand fire effluents; be simple to operate; allow individual bubbler times to be pre-set; be capable of being controlled remotely. To test the analysers functionality, it was used alongside the ISO/TS 19700 Steady State Tube Furnace (SSTF). A series of tests were conducted to assess the validity of the box analyser measurements and the data logging abilities of the apparatus. PMMA and PA 6.6 were used to assess the validity of the box analyser measurements. The data obtained from the bench-scale assessments showed excellent agreement. Following this, the portable analyser was used to monitor gas concentrations during large-scale testing using the ISO 9705 room corner test. The analyser was set up, calibrated and set to record smoke toxicity measurements in the doorway of the test room. The analyser was successful in operating without manual interference and successfully recorded data for 12 of the 12 tests conducted in the ISO room tests. At the end of each test, the analyser created a data file (formatted as .csv) containing the measured gas concentrations throughout the test, which do not require specialist knowledge to interpret. This validated the portable analyser’s ability to monitor fire effluent without operator intervention on both a bench and large-scale. The portable analyser is a validated and significantly more practical alternative to FTIR, proven to work for large-scale fire testing for quantification of smoke toxicity. The analyser is a cheaper, more accessible option to assess smoke toxicity, mitigating the need for expensive equipment and specialist operators.

Keywords: smoke toxicity, large-scale tests, iso 9705, analyser, novel equipment

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Authors: Iñaki Iturria, Pasquale Russo, Montserrat Nacher-Vázquez, Giuseppe Spano, Paloma López, Miguel Angel Pardo

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Introduction: It is known that some Lactic Acid Bacteria (LAB) present an interesting probiotic effect. Probiotic bacteria stimulate host resistance to microbial pathogens and thereby aid in immune response, and modulate the host's immune responses to antigens with a potential to down-regulate hypersensitivity reactions. Therefore, probiotic therapy is valuable against intestinal infections and may be beneficial in the treatment of Inflammatory Bowel Disease (IBD). Several in vitro tests are available to evaluate the probiotic potential of a LAB strain. However, an in vivo model is required to understand the interaction between the host immune system and the bacteria. During the last few years, zebrafish (Danio rerio) has gained interest as a promising vertebrate model in this field. This organism has been extensively used to study the interaction between the host and the microbiota, as well as the host immune response under several microbial infections. In this work, we report on the use of the zebrafish model to investigate in vivo the colonizing ability and the immunomodulatory effect of probiotic LAB. Methods: Lactobacillus strains belonging to different LAB species were fluorescently tagged and used to colonize germ-free zebrafish larvae gastrointestinal tract (GIT). Some of the strains had a well-documented probiotic effect (L. acidophilus LA5); while others presented an exopolysaccharide (EPS) producing phenotype, thus allowing evaluating the influence of EPS in the colonization and immunomodulatory effect. Bacteria colonization was monitored for 72 h by direct observation in real time using fluorescent microscopy. CFU count per larva was also evaluated at different times. The immunomodulatory effect was assessed analysing the differential expression of several innate immune system genes (MyD88, NF-κB, Tlr4, Il1β and Il10) by qRT- PCR. The anti-inflammatory effect was evaluated using a chemical enterocolitis zebrafish model. The protective effect against a pathogen was also studied. To that end, a challenge test was developed using a fluorescently tagged pathogen (Vibrio anguillarum-GFP+). The progression of the infection was monitored up to 3 days using a fluorescent stereomicroscope. Mortality rates and CFU counts were also registered. Results and conclusions: Larvae exposed to EPS-producing bacteria showed a higher fluorescence and CFU count than those colonized with no-EPS phenotype LAB. In the same way, qRT-PCR results revealed an immunomodulatory effect on the host after the administration of the strains with probiotic activity. A downregulation of proinflammatory cytoquines as well as other cellular mediators of inflammation was observed. The anti-inflammatory effect was found to be particularly marked following exposure to LA% strain, as well as EPS producing strains. Furthermore, the challenge test revealed a protective effect of probiotic administration. As a matter of fact, larvae fed with probiotics showed a decrease in the mortality rate ranging from 20 to 35%. Discussion: In this work, we developed a promising model, based on the use of gnotobiotic zebrafish coupled with a bacterial fluorescent tagging in order to evaluate the probiotic potential of different LAB strains. We have successfully used this system to monitor in real time the colonization and persistence of exogenous LAB within the gut of zebrafish larvae, to evaluate their immunomodulatory effect and for in vivo competition assays. This approach could bring further insights into the complex microbial-host interactions at intestinal level.

Keywords: gnotobiotic, immune system, lactic acid bacteria, probiotics, zebrafish

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