Search results for: Petr Saha

Authors: Matej Patzelt, Jan Dudak, Frantisek Krejci, Jan Zemlicka, Vladimir Musil, Jitka Riedlova, Viktor Sykora, Jana Mrzilkova, Petr Zach

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Introduction: Micro-CT is well used for examination of bone structures and teeth. On the other hand visualization of the soft tissues is still limited. The goal of our study was to create a new fixation method for soft tissue imaging in micro-CT. Methodology: We used organs of 18 mice - heart, lungs, kidneys, liver and brain, which we fixated in ethanol after meticulous preparation. We fixated organs in different concentrations of ethanol and for different period of time. We used three types of ethanol concentration - 97%, 50% and ascending ethanol concentration (25%, 50%, 75%, 97% each for 12 hours). Fixated organs were scanned after 72 hours, 168 hours and 336 hours period of fixation. We scanned all specimens in micro-CT MARS (Medipix All Resolution System). Results: Ethanol method provided contrast enhancement in all studied organs in all used types of fixation. Fixation in 97% ethanol provided very fast fixation and the contrast among the tissues was visible already after 72 hours of fixation. Fixation for the period of 168 and 336 hours gave better details, especially in lung tissue, where alveoli were visualized. On the other hand, this type of fixation caused organs to petrify. Fixation in 50% ethanol provided best results in 336 hours fixation, details were visualized better than in 97% ethanol and samples were not as hard as in fixation in 97% ethanol. Best results were obtained in fixation in ascending ethanol concentration. All organs were visualized in great details, best-visualized organ was heart, where trabeculae and valves were visible. In this type of fixation, organs stayed soft for whole time. Conclusion: New ethanol method is a great option for soft tissue fixation as well as the method for enhancing contrast among tissues in organs. The best results were obtained with fixation of the organs in ascending ethanol concentration, the best visualized organ was the heart.

Keywords: x-ray imaging, small animals, ethanol, ex-vivo

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Authors: Radka Obořilová, Hana Šimečková, Matěj Pastucha, Jan Přibyl, Petr Skládal, Ivana Mašlaňová, Zdeněk Farka

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Due to the spreading of antimicrobial resistance, alternative approaches to combat superinfections are being sought, both in the field of lysing agents and methods for studying bacterial lysis. A suitable alternative to antibiotics is phage therapy and enzybiotics, for which it is also necessary to study the mechanism of their action. Biosensor-based techniques allow rapid detection of pathogens in real time, verification of sensitivity to commonly used antimicrobial agents, and selection of suitable lysis agents. The detection of lysis takes place on the surface of the biosensor with immobilized bacteria, which has the potential to be used to study biofilms. An example of such a biosensor is surface plasmon resonance (SPR), which records the kinetics of bacterial lysis based on a change in the resonance angle. The bacteria are immobilized on the surface of the SPR chip, and the action of phage as the mass loss is monitored after a typical lytic cycle delay. Atomic force microscopy (AFM) is a technique for imaging of samples on the surface. In contrast to electron microscopy, it has the advantage of real-time imaging in the native conditions of the nutrient medium. In our case, Staphylococcus aureus was lysed using the enzyme lysostaphin and phage P68 from the familyPodoviridae at 37 ° C. In addition to visualization, AFM was used to study changes in mechanical properties during lysis, which resulted in a reduction of Young’s modulus (E) after disruption of the bacterial wall. Changes in E reflect the stiffness of the bacterium. These advanced methods provide deeper insight into bacterial lysis and can help to fight against bacterial diseases.

Keywords: biosensors, atomic force microscopy, surface plasmon resonance, bacterial lysis, staphylococcus aureus, phage P68

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Authors: Ashish Bohre, Blaž Likozar, Saikat Dutta, Dionisios G. Vlachos, Basudeb Saha

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Graphene oxide, decorated with surface oxygen functionalities, has emerged as a sustainable alternative to precious metal catalysts for many reactions. Herein, we report for the first time that graphene oxide becomes super active for C-C coupling upon incorporation of multilayer crystalline features, highly oxidized surface, Brønsted acidic functionalities and defect sites on the surface and edges via modified oxidation. The resulting improved graphene oxide (IGO) demonstrates superior activity to commonly used framework zeolites for upgrading of low carbon biomass furanics to long carbon chain aviation fuel precursors. A maximum 95% yield of C15 fuel precursor with high selectivity is obtained at low temperature (60 C) and neat conditions via hydroxyalkylation/alkylation (HAA) of 2-methylfuran (2-MF) and furfural. The coupling of 2-MF with carbonyl molecules ranging from C3 to C6 produced the precursors of carbon numbers 12 to 21. The catalyst becomes inactive in the 4th cycle due to the loss of oxygen functionalities, defect sites and multilayer features; however, regains comparable activity upon regeneration. Extensive microscopic and spectroscopic characterization of the fresh and reused IGO is presented to elucidate high activity of IGO and to establish a correlation between activity and surface and structural properties. Kinetic Monte Carlo (KMC) and density functional theory (DFT) calculations are presented to further illustrate the surface features and the reaction mechanism.

Keywords: methacrylic acid, itaconic acid, biomass, monomer, solid base catalyst

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Authors: Harish Rajak, Kamlesh Raghuwanshi

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Histone deacetylase (HDAC) are promising target for cancer treatment. The panobinostat (Farydak; Novartis; approved by USFDA in 2015) and chidamide (Epidaza; Chipscreen Biosciences; approved by China FDA in 2014) are the novel HDAC inhibitors ratified for the treatment of patients with multiple myeloma and peripheral T cell lymphoma, respectively. On the other hand, two other HDAC inhibitors, Vorinostat (SAHA; approved by USFDA in 2006) and Romidepsin (FK228; approved by USFDA in 2009) are already in market for the treatment of cutaneous T-cell lymphoma. Several hydroxamic acid based HDAC inhibitors i.e., belinostat, givinostat, PCI24781 and JNJ26481585 are in clinical trials. HDAC inhibitors consist of three pharmacophoric features - an aromatic cap group, zinc binding group (ZBG) and a linker chain connecting cap group to ZBG. Herein, we report synthesis, characterization and biological evaluation of HDAC inhibitors possessing substituted isatin moiety as cap group which recognize the surface of active enzyme pocket and thiosemicarbazide moiety incorporated as linker group responsible for connecting cap group to ZBG (hydroxamic acid). Several analogues were found to inhibit HDAC and cellular proliferation of Hela cervical cancer cells with GI50 values in the micro molar range. Some of the compounds exhibited promising results in vitro antiproliferative studies. Attempts were also made to establish the structure activity relationship among synthesized HDAC inhibitors.

Keywords: HDAC inhibitors, hydroxamic acid derivatives, isatin derivatives, antiproliferative activity, docking

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Authors: Radek Richtr, Petr Pauš

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Social media nowadays connects people more tightly and intensively than ever, but simultaneously, some sort of social distance, incomprehension, lost of social integrity appears. People can be strongly connected to the person on the other side of the world but unaware of neighbours in the same district or street. The Stoa is a type of application from the ”serious games” genre- it is research augmented reality experiment masked as a gaming environment. In the Stoa environment, the player can plant and grow virtual (organic) structure, a Pillar, that represent the whole suburb. Everybody has their own idea of what is an acceptable, admirable or harmful visual intervention in the area they live in; the purpose of this research experiment is to find and/or define residents shared subconscious spirit, genius loci of the Pillars vicinity, where residents live in. The appearance and evolution of Stoa’s Pillars reflect the real world as perceived by not only the creator but also by other residents/players, who, with their actions, refine the environment. Squares, parks, patios and streets get their living avatar depictions; investors and urban planners obtain information on the occurrence and level of motivation for reshaping the public space. As the project is in product conceptual design phase, the function is one of its most important factors. Function-based modelling makes design problem modular and structured and thus decompose it into sub-functions or function-cells. Paper discuss the current conceptual model for Stoa project, the using of different organic structure textures and models, user interface design, UX study and project’s developing to the final state.

Keywords: augmented reality, urban computing, interaction design, mixed reality, social engineering

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Authors: Prodipta Bhowmik, Poulami Saha, Preety Mehra, Yogesh Soni, Triloki Nath Jha

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In India, the smart city concept is growing day by day. So, for smart city development, a better traffic management and monitoring system is a very important requirement. Nowadays, road accidents increase due to more vehicles on the road. Reckless driving is mainly responsible for a huge number of accidents. So, an efficient traffic management system is required for all kinds of roads to control the traffic speed. The speed limit varies from road to road basis. Previously, there was a radar system but due to high cost and less precision, the radar system is unable to become favorable in a traffic management system. Traffic management system faces different types of problems every day and it has become a researchable topic on how to solve this problem. This paper proposed a computer vision and machine learning-based automated system for multiple vehicle detection, tracking, and speed estimation of vehicles using image processing. Detection of vehicles and estimating their speed from a real-time video is tough work to do. The objective of this paper is to detect vehicles and estimate their speed as accurately as possible. So for this, a real-time video is first captured, then the frames are extracted from that video, then from that frames, the vehicles are detected, and thereafter, the tracking of vehicles starts, and finally, the speed of the moving vehicles is estimated. The goal of this method is to develop a cost-friendly system that can able to detect multiple types of vehicles at the same time.

Keywords: OpenCV, Haar Cascade classifier, DLIB, YOLOV3, centroid tracker, vehicle detection, vehicle tracking, vehicle speed estimation, computer vision

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Authors: Arup Saha Chaudhuri

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Structural steel is a very ductile material with high strength carrying capacity, thus it is very useful to make earthquake resistant buildings. It is a homogeneous material also. The member section and the structural system can be made very efficient for economical design. As the steel is recyclable and reused, it is a green material. The embodied energy for the efficiently designed steel structure is less than the RC structure. For sustainable green building steel is the best material nowadays. Moreover, pre-engineered and pre-fabricated faster construction methodologies help the development work to complete within the stipulated time. In this paper, the usefulness of Eccentric Bracing Frame (EBF) in steel structure over Moment Resisting Frame (MRF) and Concentric Bracing Frame (CBF) is shown. Stability of the steel structures against horizontal forces especially in seismic condition is efficiently possible by Eccentric bracing systems with economic connection details. The EBF is pin–ended, but the beam-column joints are designed for pin ended or for full connectivity. The EBF has several desirable features for seismic resistance. In comparison with CBF system, EBF system can be designed for appropriate stiffness and drift control. The link beam is supposed to yield in shear or flexure before initiation of yielding or buckling of the bracing member in tension or compression. The behavior of a 2-D steel frame is observed under seismic loading condition in the present paper. Ductility and brittleness of the frames are compared with respect to time period of vibration and dynamic base shear. It is observed that the EBF system is better than MRF system comparing the time period of vibration and base shear participation.

Keywords: steel building, green and sustainable, earthquake resistant, EBF system

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Authors: András Farkas, István Molnár, Jan Vrána, Veronika Burešová, Petr Cápal, András Cseh, Márta Molnár-Láng, Jaroslav Doležel

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Species of Aegilops played a central role in the evolution of wheat and are sources of traits related to yield quality and tolerance against biotic and abiotic stresses. These wild genes and alleles are desirable to use in crop improvement programs via introgressive hybridization. However, the success of chromosome mediated gene transfer to wheat are hampered by the pour knowledge on the genome structure of Aegilops relative to wheat and by the low number of cost-effective molecular markers specific for Aegilops chromosomes. The COS markers specific for genes conserved throughout evolution in both sequence and copy number between Triticeae/Aegilops taxa and define orthologous regions, thus enabling the comparison of regions on the chromosomes of related species. The present study compared individual chromosomes of Aegilops umbellulata (UU), Ae. comosa (MM), Ae. speltoides (SS) and Ae. caudata (CC) purified by flourescent labelling with oligonucleotid SSR repeats and biparametric flow cytometry with wheat by identifying orthologous chromosomal regions by COS markers. The linear order of bin-mapped COS markers along the wheat D chromosomes was identified by the use of chromosome-specific sequence data and virtual gene order. Syntenic regions of wheat identifying genome rearrangements differentiating the U, M, S or C genomes from the D genome of wheat were detected. The conserved orthologous set markers assigned to Aegilops chromosomes promise to accelerate gene introgression by facilitating the identification of alien chromatin. The syntenic relationships between the Aegilops species and wheat will facilitate the targeted development of new markers specific for U, M, S and C genomic regions and will contribute to the understanding of molecular processes related to the evolution of Aegilops.

Keywords: Aegilops, cos-markers, flow-sorting, wheat

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Authors: Jan Luxa, Vlastimil Mazanek, Petr Malinsky, Alexander Romanenko, Mariapompea Cutroneo, Vladimir Havranek, Josef Novak, Eva Stepanovska, Anna Mackova, Zdenek Sofer

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Using accelerated energetic ions is an interesting method for the introduction of structural changes in various carbon-based materials. This way, the properties can be altered in two ways: a) the ions lead to the formation of conductive pathways in graphene oxide structures due to the elimination of oxygen functionalities and b) doping with selected ions to form metal nanoclusters, thus increasing the conductivity. In this work, energetic beams were employed in two ways to prepare capacitor structures in graphene oxide (GO), reduced graphene oxide (rGO) and polyimide (PI) on a micro-scale. The first method revolved around using ion beam writing with a focused ion beam, and the method involved ion implantation via a polymeric mask. To prepare the polymeric mask, a direct spin-coating of PMMA on top of the foils was used. Subsequently, proton beam writing and development in isopropyl alcohol were employed. Finally, the mask was removed using acetone solvent. All three materials were exposed to ion beams with an energy of 2.5-5 MeV and an ion fluence of 3.75x10¹⁴ cm-² (1800 nC.mm-²). Thus, prepared microstructures were thoroughly characterized by various analytical methods, including Scanning electron microscopy (SEM) with Energy-Dispersive X-ray spectroscopy (EDS), X-ray Photoelectron spectroscopy (XPS), micro-Raman spectroscopy, Rutherford Back-scattering Spectroscopy (RBS) and Elastic Recoil Detection Analysis (ERDA) spectroscopy. Finally, these materials were employed and tested as sensors for humidity using electrical conductivity measurements. The results clearly demonstrate that the type of ions, their energy and fluence all have a significant influence on the sensory properties of thus prepared sensors.

Keywords: graphene, graphene oxide, polyimide, ion implantation, sensors

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Authors: Tesfay Gidey, Emiru Birhane, Ashenafi Manaye, Hailemariam Kassa, Tesfay Atsbha, Negasi Solomon, Hadgu Hishe, Aklilu Negussie, Petr Madera, Jose G. Borges

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The globally endangered Dracaena ombet is one of the ten dragon multipurpose tree species in arid ecosystems. Anthropogenic and natural factors are now impacting the sustainability of the species. This study was conducted to prioritize criteria and alternative strategies for the conservation of the species using the analytical hierarchy process (AHP) model by involving all relevant stakeholders in the Desa'a dry Afromontane forest in northern Ethiopia. Information about the potential alternative strategies and the criteria for their evaluation was first collected from experts, personal experiences, and literature reviews. Afterward, they were validated using stakeholders' focus group discussions. Five candidate strategies with three evaluation criteria were considered for prioritization using the AHP techniques. The overall priority ranking value of the stakeholders showed that the ecological criterion was deemed as the most essential factor for the choice of alternative strategies, followed by the economic and social criteria. The minimum cut-off strategy, combining exclosures with the collection of only 5% of plant parts from the species, soil and water conservation, and silviculture interventions, was selected as the best alternative strategy for sustainable D. ombet conservation. The livelihood losses due to the selected strategy should be compensated by the collection of non-timber forest products, poultry farming, home gardens, rearing small ruminants, beekeeping, and agroforestry. This approach may be extended to study other dragon tree species and explore strategies for the conservation of other arid ecosystems.

Keywords: conservation strategies, analytical hierarchy process model, Desa'a forest, endangered species, Ethiopia, overexploitation

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Authors: Al-Hindi Rashad, Alotibi Ibrahim

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The aim of this study was to examine the antibacterial activity of the peroxide components of some locally produced honeys: Toran, Zaitoon (Olive), Shaflah, Saha, Jizan, Rabea Aja, Fakhira, Sedr Aljanoob, Tenhat, Karath and Bareq against two of the drug resistant bacteria; i.e., methicillin resistant Staph. aureus (MRSA, ATCC 43330) and Acinetobacter baumannii. Measurement of the antibacterial activity of honey samples by using the agar well diffusion method was adopted as follows: by using turbidity standard McFaraland 0.5, suspensions of bacterial strains MRSA ATCC 43330 and Acinetobacter baumannii were prepared. By the spreading plate method, 100 µl of the suspension was inoculated onto Muller-Hinton agar medium. On the inoculated agar medium, five wells were made using a sterile cork borer (diameter 5 mm).100 µl of honey dilutions (10%, 30%, 50%, 70% and 100%) were used. The study indicated that the highly effective activity was in some local honey samples such as Toran honey against MRSA, and Shafalah honey against MRSA and Acinetobacter baumannii which showed bactericidal effects at concentrations 70 % to 100 % as well. The majority of local honey samples recorded bacteriostatic effects on MRSA and Acinetobacter baumannii at consternations 50 % and above. In conclusion this investigation indicated that in regard to the majority inhibitory effect on microorganisms, the existing of H2O2 in honey samples together with phenolic content greatly provide a strong antibacterial activities among different types of honey, because in some previous studies the H2O2 content of honey interacts with phenolic content and showed better inhibitory effect than in absent of H2O2.

Keywords: antibacterial activity, honey, hospital acquired, Saudi Arabia

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Authors: Vojtech Stejskal, Hana Skalova, Petr Kvapil, George Hutchinson

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Algae growth has been an important issue in water management of water plants, ponds and lakes, swimming pools, aquaculture & fish farms, gardens or golf courses for last decades. There are solutions based on chemical or biological principles. Apart of these traditional principles for inhibition of algae growth and biofilm production there are also physical methods which are very competitive compared to the traditional ones. Ultrasonic technology is one of these alternatives. Ultrasonic emitter is able to eliminate the biofilm which behaves as a host and attachment point for algae and is original reason for the algae growth. The ultrasound waves prevent majority of the bacteria in planktonic form becoming strongly attached sessile bacteria that creates welcoming layer for the biofilm production. Biofilm creation is very fast – in the serene water it takes between 30 minutes to 4 hours, depending on temperature and other parameters. Ultrasound device is not killing bacteria. Ultrasound waves are passing through bacteria, which retract as if they were in very turbulent water even though the water is visually completely serene. In these conditions, bacteria does not excrete the polysaccharide glue they use to attach to the surface of the pool or pond, where ultrasonic technology is used. Ultrasonic waves decrease the production of biofilm on the surfaces in the selected area. In case there are already at the start of the application of ultrasonic technology in a pond or basin clean inner surfaces, the biofilm production is almost absolutely inhibited. This paper talks about two different pilot applications – one in Czech Republic and second in United States of America, where the used ultrasonic technology (AlgaeControl) is coming from. On both sites, there was used Mezzo Ultrasonic Algae Control System with very positive results not only on biofilm production, but also algae growth in the surrounding area. Technology has been successfully tested in two different environments. The poster describes the differences and their influence on the efficiency of ultrasonic technology application. Conclusions and lessons learned can be possibly applied also on other sites within Europe or even further.

Keywords: algae growth, biofilm production, ultrasonic solution, ultrasound

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Authors: Vishnu A., Ashesh Saha

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The operation of high-speed rotating machinery in industries is accompanied by rotor vibrations due to many factors. One of the primary instability mechanisms in a rotor system is the centrifugal force induced due to the eccentricity of the center of mass away from the center of rotation. These unwanted vibrations may lead to catastrophic fatigue failure. So, there is a need to control these rotor vibrations. In this work, control of rotor vibrations by using a 4-pole Radial Active Magnetic Bearing (RAMB) as an actuator is analysed. A continuous rotor system model is considered for the analysis. Several important factors, like the gyroscopic effect and rotary inertia of the shaft and disc, are incorporated into this model. The large deflection of the shaft and the restriction to axial motion of the shaft at the bearings result in nonlinearities in the system governing equation. The rotor system is modeled in such a way that the system dynamics can be related to the geometric and material properties of the shaft and disc. The mathematical model of the rotor system is developed by incorporating the control forces generated by the RAMB. A simple PD controller is used for the attenuation of system vibrations. An analytical expression for the amplitude and phase equations is derived using the Method of Multiple Scales (MMS). Analytical results are verified with the numerical results obtained using an ‘ode’ solver in-built into MATLAB Software. The control force is found to be effective in attenuating the system vibrations. The multi-valued solutions leading to the jump phenomenon are also eliminated with a proper choice of control gains. Most interestingly, the shape of the backbone curves can also be altered for certain values of control parameters.

Keywords: rotor dynamics, continuous rotor system model, active magnetic bearing, PD controller, method of multiple scales, backbone curve

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Authors: Bishal Saha, Musah Ahmed Rufai Muhyedeen, Jubeyer Hossain Joy, Muhammad Muhitur Rahman, Mohammad Shahedur Rahman, Md Arif Hasan, Syed Masiur Rahman

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Greenhouse gas (GHG) emission is one of the critical problems of today’s world. Many countries have been taking many short- and long-term plans to reduce climate change mitigation. However, the potential of behavioral changes in addressing this problem is promising, as reported by many researchers. This paper presents a comprehensive literature review that focuses on ways to influence people’s behavior in their homes, workplace, and transportation to mitigate the emission directly or indirectly. This study will investigate different theories pertinent to planned behavior and the key elements for modifying behavior like biophilia, reinforcement to use optimum energy and recyclable products, proper application of greenhouse tax, modern technology, and sustainable design adaptation, transportation sharing, social and community norms, proper education and information, and financial incentives. There is a number of challenges associated with behavioral changes. Behavioral interventions have different actions varied by their type and need to combine various policy tools and great social marketing. Many interventions can reduce GHG emissions without any compromise with household well-being. This study will develop a landscape of prevailing theories of environmental psychology by identifying and reviewing the key themes and findings of this field of study. It will support especially the developing countries to reduce GHG emissions without significant capital investment. It is also expected that the behavioral changes will lead to the successful adoption of climate-friendly policies easily. This study will also generate new research questions and directions.

Keywords: behavioral changes, climate change mitigation, environmental psychology, greenhouse gas emission

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Authors: Paul Narh Doku

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The relationship between parental HIV/AIDS status or death and child mental health is well known, although the role of child maltreatment as a confounder or mediator in this relationship remains uncertain. This study examined the potential path mechanism through child maltreatment mediating the link between HIV/AIDS family dysfunction trajectories and psychosocial problems. A cross-sectional survey was conducted in the Lower Manya Municipal Assembly of Ghana. A questionnaire which consisted of the Strengths and Difficulties Questionnaire (SDQ), Social and Health Assessment (SAHA), Rosenberg Self-Esteem Scale (RSES), and the Conflict Tactics Scale (CTS) was completed by 291 adolescents. Controlling for relevant sociodemographic confounders, mediation analyses using linear regression were fitted to examine whether the association between family dysfunction and psychosocial problems is mediated by child maltreatment. The results indicate that, among adolescents, child maltreatment fully mediated the association between being orphaned by AIDS and self-esteem, delinquency and risky behaviours, and peer problems. Similarly, child maltreatment fully mediated the association between living with an HIV/AIDS-infected parent and self-esteem, delinquency and risky behaviours, depression/emotional problems, and peer problems. Partial mediation was found for hyperactivity. Child maltreatment mediates the association between the family dysfunction trajectories of parental HIV/AIDS or death and psychosocial problems among adolescents. This implies that efforts to address child maltreatment among families affected by HIV/AIDS may be helpful in the prevention of psychosocial problems among these children, thus enhancing their well-being. The findings, therefore, underscore the need for comprehensive psychosocial interventions that address both the unique negative exposures of HIV/AIDS and maltreatment for children affected by HIV.

Keywords: child maltreatment, child abuse, mental health, psychosocial problems, domestic violence, HIV/AIDS, adolescents

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Authors: Esha Saha, Pradip Kumar Ray

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A hospital inventory comprises of a large number and great variety of items for the proper treatment and care of patients, such as pharmaceuticals, medical equipment, surgical items, etc. Improper management of these items, i.e. stockouts, may lead to delay in treatment or other fatal consequences, even death of the patient. So, generally the hospitals tend to overstock items to avoid the risk of stockout which leads to unnecessary investment of money, difficulty in storing, more expiration and wastage, etc. Thus, in such challenging environment, it is necessary for hospitals to follow an inventory policy considering the stochasticity of demand in a hospital. Statistical analysis captures the correlation of patient condition based on bed occupancy with the patient demand which changes stochastically. Due to the dependency on bed occupancy, the markov model is developed that helps to map the changes in demand of hospital inventory based on the changes in the patient condition represented by the movements of bed occupancy states (acute care state, rehabilitative state and long-care state) during the length-of-stay of patient in a hospital. An inventory policy is developed for a hospital based on the fulfillment of patient demand with the objective of minimizing the frequency and quantity of placement of orders of inventoried items. The analytical structure of the model based on probability calculation is provided to show the optimal inventory-related decisions. A case-study is illustrated in this paper for the development of hospital inventory model based on patient demand for multiple inpatient pharmaceutical items. A sensitivity analysis is conducted to investigate the impact of inventory-related parameters on the developed optimal inventory policy. Therefore, the developed model and solution approach may help the hospital managers and pharmacists in managing the hospital inventory in case of stochastic demand of inpatient pharmaceutical items.

Keywords: bed occupancy, hospital inventory, markov model, patient condition, pharmaceutical items

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Authors: Josef Fládr, Jiří Němeček, Veronika Koudelková, Petr Bílý

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Mechanical parameters of cementitious composites differ quite significantly based on the composition of cement matrix. They are also influenced by mixing times and procedure. The research presented in this paper was aimed at identification of differences in microstructure of normal strength (NSC) and differently mixed high strength (HSC) cementitious composites. Scanning electron microscopy (SEM) investigation together with energy dispersive X-ray spectroscopy (EDX) phase analysis of NSC and HSC samples was conducted. Evaluation of interfacial transition zone (ITZ) between the aggregate and cement matrix was performed. Volume share, thickness, porosity and composition of ITZ were studied. In case of HSC, samples obtained by several different mixing procedures were compared in order to find the most suitable procedure. In case of NSC, ITZ was identified around 40-50% of aggregate grains and its thickness typically ranged between 10 and 40 µm. Higher porosity and lower share of clinker was observed in this area as a result of increased water-to-cement ratio (w/c) and the lack of fine particles improving the grading curve of the aggregate. Typical ITZ with lower content of Ca was observed only in one HSC sample, where it was developed around less than 15% of aggregate grains. The typical thickness of ITZ in this sample was similar to ITZ in NSC (between 5 and 40 µm). In the remaining four HSC samples, no ITZ was observed. In general, the share of ITZ in HSC samples was found to be significantly smaller than in NSC samples. As ITZ is the weakest part of the material, this result explains to large extent the improved mechanical properties of HSC compared to NSC. Based on the comparison of characteristics of ITZ in HSC samples prepared by different mixing procedures, the most suitable mixing procedure from the point of view of properties of ITZ was identified.

Keywords: electron diffraction spectroscopy, high strength concrete, interfacial transition zone, normal strength concrete, scanning electron microscopy

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Authors: Arijit Saha, Hassan Kassem, Leo Hoening

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Ongoing climate change demands the increasing use of renewable energies. Wind energy plays an important role in this context since it can be applied almost everywhere in the world. To reduce the costs of wind turbines and to make them more competitive, simulations are very important since experiments are often too costly if at all possible. The wind turbine on a vast open area experiences the turbulence generated due to the atmosphere, so it was of utmost interest from this research point of view to generate the turbulence through various Inlet Turbulence Generation methods like Precursor cyclic and Kaimal Spectrum Exponential Coherence (KSEC) in the computational simulation domain. To be able to validate computational fluid dynamic simulations of wind turbines with the experimental data, it is crucial to set up the conditions in the simulation as close to reality as possible. This present work, therefore, aims at investigating the turbulent inflow strategy and boundary conditions of KSEC and providing a comparative analysis alongside the Precursor cyclic method for Large Eddy Simulation within the context of wind energy applications. For the generation of the turbulent box through KSEC method, firstly, the constrained data were collected from an auxiliary channel flow, and later processing was performed with the open-source tool PyconTurb, whereas for the precursor cyclic, only the data from the auxiliary channel were sufficient. The functionality of these methods was studied through various statistical properties such as variance, turbulent intensity, etc with respect to different Bulk Reynolds numbers, and a conclusion was drawn on the feasibility of KSEC method. Furthermore, it was found necessary to verify the obtained data with DNS case setup for its applicability to use it as a real field CFD simulation.

Keywords: Inlet Turbulence Generation, CFD, precursor cyclic, KSEC, large Eddy simulation, PyconTurb

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Authors: Diptiman Dinda, Shyamal Kumar Saha

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In the 21st century, sensitive and selective detection of trace amounts of explosives has become a serious problem. Generally, nitro compound and its derivatives are being used worldwide to prepare different explosives. Recently, TNP (2, 4, 6 trinitrophenol) is the most commonly used constituent to prepare powerful explosives all over the world. It is even powerful than TNT or RDX. As explosives are electron deficient in nature, it is very difficult to detect one separately from a mixture. Again, due to its tremendous water solubility, detection of TNP in presence of other explosives from water is very challenging. Simple instrumentation, cost-effective, fast and high sensitivity make fluorescence based optical sensing a grand success compared to other techniques. Graphene oxide (GO), with large no of epoxy grps, incorporate localized nonradiative electron-hole centres on its surface to give very weak fluorescence. In this work, GO is functionalized with 2, 6-diamino pyridine to remove those epoxy grps. through SN2 reaction. This makes GO into a bright blue luminescent fluorophore (DAP/rGO) which shows an intense PL spectrum at ∼384 nm when excited at 309 nm wavelength. We have also characterized the material by FTIR, XPS, UV, XRD and Raman measurements. Using this as fluorophore, a large fluorescence quenching (96%) is observed after addition of only 200 µL of 1 mM TNP in water solution. Other nitro explosives give very moderate PL quenching compared to TNP. Such high selectivity is related to the operation of FRET mechanism from fluorophore to TNP during this PL quenching experiment. TCSPC measurement also reveals that the lifetime of DAP/rGO drastically decreases from 3.7 to 1.9 ns after addition of TNP. Our material is also quite sensitive to 125 ppb level of TNP. Finally, we believe that this graphene based luminescent material will emerge a new class of sensing materials to detect trace amounts of explosives from aqueous solution.

Keywords: graphene, functionalization, fluorescence quenching, FRET, nitroexplosive detection

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Authors: Saha Naseri

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Introduction/Objective: Son preference, a preference for sons over daughters, is a practice deeply-rooted in gender inequality that is widespread in many societies and across different religions and cultures. In this study, we are aiming to study the effects of son preference on fertility outcomes (birth interval and current contraceptive use) in Afghanistan, where have been perceived with high rates of son preference. The objectives of the study are to examine the association between the sex of the previous child and the duration of the subsequent birth interval and to evaluate the effect of son preference on current contraceptive use. Methodology: Afghanistan Demographic and Health Survey (DHS) (2015) was used to study the impact of son preference on fertility outcomes among married women. The data collected from 28,661 on currently-married women, aged between 15 and 49 years who have at least one child, have used to conduct this quantitative study. Outcomes of interest are birth interval and current contraceptive use. Simple and multiple regression analysis have been conducted to assess the effect of son preference on these fertility outcomes. Results: The present study has highlighted that son preference somehow exists among married women in Afghanistan. It is indicated that the sex of the first birth is significantly associated with the succeeding birth interval. Having a female child as the first baby was associated with a shorter average succeeding birth interval by 1.8 months compared to a baby boy (p-value = 0.000). For the second model, the results identified that women who desire for more sons have 7% higher odds to be current contraceptive user compared to those who have no preference (p-value = 0.03). The latter results do not indicate the son preference. However, one limitation for this result was the timeliness of the questions asked since contraceptive use in the current time was asked along with a question on ‘future’ desired sex composition. Moreover, women may have just given birth and want to reach a certain time span of birth interval before planning for another child, even if it was a boy, which might have affected the results. Conclusion: Overall, this study has demonstrated that there is a positive relationship between son preference and one main fertility behaviors, birth interval. The second fertility outcome, current contraceptive use, was not a good indicator to measure son preference. Based on the finding, recommendations will be made for appropriate interventions addressing gender norms and related fertility decisions.

Keywords: Afghanistan, birth interval, contraceptive, son preference

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Authors: Matej Patzelt, Jana Mrzilkova, Jan Dudak, Frantisek Krejci, Jan Zemlicka, Zdenek Wurst, Petr Zach, Vladimir Musil

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Introduction: Micro-CT is well used for examination of bone structures and teeth. On the other hand visualization of the soft tissues is still limited. The goal of our study was to elaborate methodology for soft tissue samples imaging in micro-CT. Methodology: We used organs of rats and mice. We either did a preparation of the organs and fixation in contrast solution or we did cannulation of blood vessels and their injection for imaging of the vascular system. First, we scanned native specimens, then we created corrosive specimens by resins. In the next step, we injected vascular system either by Aurovist contrast agent or by Exitron. In the next step, we focused on soft tissues contrast increase. We scanned samples fixated in Lugol solution, samples fixated in pure ethanol and in formaldehyde solution. All used methods were afterwards compared. Results: Native specimens did not provide sufficient contrast of the tissues in any of organs. Corrosive samples of the blood stream provided great contrast and details; on the other hand, it was necessary to destroy the organ. Further examined possibility was injection of the AuroVist contrast that leads to the great bloodstream contrast. Injection of Exitron contrast agent comparing to Aurovist did not provide such a great contrast. The soft tissues (kidney, heart, lungs, brain, and liver) were best visualized after fixation in ethanol. This type of fixation showed best results in all studied tissues. Lugol solution had great results in muscle tissue. Fixation by formaldehyde solution showed similar quality of contrast in the tissues like ethanol. Conclusion: Before imaging, we need to, first, determinate which structures of the soft tissues we want to visualize. In the case of the bloodstream, the best was AuroVist and corrosive specimens. Muscle tissue is best visualized by Lugol solution. In the case of the organs containing cavities, like kidneys or brain, the best way was ethanol fixation.

Keywords: experimental imaging, fixation, micro-CT, soft tissues

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Authors: Konstantin Yu. Moiseev, Antonina F. Budnik, Andrey I. Emanuilov, Petr M. Masliukov

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The vast majority of sympathetic ganglionic neurons are catecholaminergic. Some sympathetic neurons lack catecholamines and mostly use acetylcholine as their main neurotransmitter. Some cholinergic postganglionic neurons also express neuronal nitric oxide synthase (nNOS). Preganglionic sympathetic neurons are cholinergic and most of them are also nNOS-immunoreactive (IR). The purpose of this study was to gain further insight into the neuroplasticity of sympathetic neurons during postnatal ontogenesis by comparing the development of pre- and postganglionic neurons expressing nNOS in different mammals. nNOS was investigated by immunohistochemistry in the sympathetic superior cervical ganglion (SCG), stellate ganglion (SG), celiac ganglion (CG) and spinal cord from rats, mice and cats of different ages (newborn, 10-day-old, 20-day-old, 30-day-old, 2-month-old and 2-year-old). In rats and mice, nNOS-positive neurons were not found in sympathetic ganglia from birth onwards. In cats, non-catecholaminergic nNOS-IR sympathetic ganglionic neurons are present from the moment of birth. In all studied age groups, substantial populations of nNOS-IR cells (up to 8.3%) was found in the SG, with a much smaller population found in the SCG (<1%) and only few cells observed in the CG. The percentage of nNOS-IR neurons in the CG and SCG did not significantly change during development. The proportion of nNOS-IR neuron profiles in the SG increased in first 20 days of life from 2.3±0.15% to 8.3±0.56%. In the SG, percentages of nNOS-IR sympathetic neurons colocalizing vasoactive intestinal peptide increased in the first 20 days of life. Choline acetyltransferase (ChAT)-IR and calcitonin gene-related peptide-IR neurons were not observed in the sympathetic ganglia of newborn animals and did not appear until 10 days after birth. In the SG of newborn and 10-day-old kittens, the majority of NOS-IR neurons were calbindin (CB)-IR, whereas in the SCG and CG of cats of all age groups and in the SG of 30-day-old and older kittens, the vast majority of NOS-IR neurons lacked CB. In newborn mammals, the most of sympathetic preganglionic neurons in the nucleus intermediolateralis thoracolumbalis pars principalis (nucl.ILp) were nNOS-IR. The percentage of nNOS-IR neurons decreased and the same parameter of ChAT-IR neurons increased during the development. We conclude that the development of nNOS-IR preganglionic and ganglionic sympathetic neurons in different mammals has time and species differences.

Keywords: sympathetic neuron, nitric oxide synthase, immunohistochemistry, development

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Authors: Jakub Vávra, Milan Valenta, Petr Kosek

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This article presents a perspective on the conduct of the dramatherapy process with persons with addictive behaviours with regard to the directiveness of the process. Although drama therapy as one of the creative arts approaches is rather non-directive in nature, depending on the clientele, there may be a need to structure the process more and, depending on the needs of the clients, to guide the process more directive. The specificity for people with addictive behaviours is discussed through the prism of the dramatherapeutic perspective, where we can find both a psychotherapeutic component as well as a component touching on expression and art, which is rather non-directive in nature. Within the context of practice with clients, this theme has repeatedly emerged and dramatherapists themselves have sought to find ways of coping with clients' demands and needs for structure and guidance within the dramatherapy process. Some of the outcomes from the supervision work also guided the research. Based on this insight, the research questions were approached. The first research question asks: in what ways is directive in dramatherapy manifested and manifested in the process? The second research question then complements the first and asks: to which phenomena are directivity in dramatherapy linked? In relation to the research questions, data were collected using focus groups and field notes. The qualitative approach of Content analysis and Relational analysis was chosen as the methodology. For analyzing qualitative research, we chose an Inductive coding scheme: Open coding, Axial coding, Pattern matching, Member checking, and Creating a coding scheme. In the presented partial research results, we find recurrent schemes related to directive coding in drama therapy. As an important element, directive leadership emerges in connection with safety for the client group, then in connection with the clients' order and also the department of the facility, and last but not least, to the personality of the drama therapist. By careful analysis and looking for patterns in the research results, we can see connections that are impossible to interpret at this stage but already provide clues to our understanding of the topic and open up further avenues for research in this area.

Keywords: dramatherapy, directivity, personal approach, aims of dramatherapy process, safetyness

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Authors: A. Saha Chaudhuri, D. Sarkar

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In steel construction, there are two families of structural members. One is hot rolled steel and another is cold formed steel. Cold formed steel section includes steel sheet, strip, plate or flat bar. Cold formed steel section is manufactured in roll forming machine by press brake or bending operation. Cold formed steel (CFS), also known as Light Gauge Steel (LGS). As cold formed steel is a sustainable material, it is widely used in green building. Cold formed steel can be recycled and reused with no degradation in structural properties. Cold formed steel structures can earn credits for green building ratings such as LEED and similar programs. Cold formed steel construction satisfies international demand for better, more efficient and affordable buildings. Cold formed steel sections are used in building, car body, railway coach, various types of equipment, storage rack, grain bin, highway product, transmission tower, transmission pole, drainage facility, bridge construction etc. Various shapes of cold formed steel sections are available, such as C section, Z section, I section, T section, angle section, hat section, box section, square hollow section (SHS), rectangular hollow section (RHS), circular hollow section (CHS) etc. In building construction cold formed steel is used as eave strut, purlin, girt, stud, header, floor joist, brace, diaphragm and covering for roof, wall and floor. Cold formed steel has high strength to weight ratio and high stiffness. Cold formed steel is non shrinking and non creeping at ambient temperature, it is termite proof and rot proof. CFS is durable, dimensionally stable and non combustible material. CFS is economical in transportation and handling. At present days cold formed steel becomes a competitive building material. In this paper all these applications related present research work are described and how the CFS can be used as blast resistant structural system that is examined.

Keywords: cold form steel sections, applications, present research review, blast resistant design

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Authors: Fariha Akhter Chowdhury, Sabrina Mahboob, Anamika Saha, Afrin Jahan, Mohammad Nurul Islam

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Urinary tract infection (UTI) is frequently experienced by the female population where the prevalence increases with aging. Escherichia coli, one of the most common UTI causing organisms, retains glutathione defense mechanism that aids the organism to withstand the harsh physiological environment of urinary tract, host oxidative immune response and even to affect antibiotic-mediated cell death and the emergence of resistance. In this study, we aimed to investigate the glutathione reductase activity of uropathogenic E. coli (UPEC) by observing the reduced glutathione (GSH) level alteration under stressful condition. Urine samples of 58 patients with UTI were collected. Upon isolation and identification, 88% of the samples presented E. coli as UTI causing organism among which randomly selected isolates (n=9), obtained from urine samples of female patients, were considered for this study. E. coli isolates were grown under normal and stressful conditions where H₂O₂ was used as the stress-inducing agent. GSH level estimation of the isolates in both conditions was carried out based on the colorimetric measurement of 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) and GSH reaction product using microplate reader assay. The GSH level of isolated E. coli sampled from adult patients decreased under stress compared to normal condition (p = 0.011). On the other hand, GSH production increased markedly in samples that were collected from elderly subjects (p = 0.024). A significant partial correlation between age and change of GSH level was found as well (p = 0.007). This study may help to reveal ways for better understanding of E. coli pathogenesis of UTI prevalence in elderly patients.

Keywords: Escherichia coli, glutathione reductase activity, oxidative stress, reduced glutathione (GSH), urinary tract infection (UTI)

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Authors: Bipul Krishna Saha, Basab Chakraborty, Ashish Alex Sam, Parthasarathi Ghosh

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The demand for energy is cumulatively increasing with time.  Since the availability of conventional energy resources is dying out gradually, significant interest is being laid on searching for alternate energy resources and minimizing the wastage of energy in various fields.  In such perspective, low-grade waste heat from several industrial sources can be reused to generate electricity. The present work is to further the adoption of the Organic Rankine Cycle (ORC) technology in Indian industrial sector.  The present paper focuses on extending the previously reported idea to the next level through a comparative review with three different working fluids using practical data from an Indian industrial plant. For comprehensive study in the simulation platform of Aspen Hysys^®, v8.6, the waste heat data has been collected from a current coke oven gas plant in India.  A parametric analysis of non-regenerative ORC and regenerative ORC is executed using the working fluids R-123, R-11 and R-21 for subcritical ORC system.  The primary goal is to determine the optimal working fluid considering various system parameters like turbine work output, obtained system efficiency, irreversibility rate and second law efficiency under applied multiple heat source temperature (160 °C- 180 °C).  Selection of the turbo-expanders is one of the most crucial tasks for low-temperature applications in ORC system. The present work is an attempt to make suitable recommendation for the appropriate configuration of the turbine. In a nutshell, this study justifies the proficiency of integrating the ORC technology in Indian perspective and also finds the appropriate parameter of all components integrated in ORC system for building up an ORC prototype.

Keywords: organic Rankine cycle, regenerative organic Rankine cycle, waste heat recovery, Indian industry

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Authors: Eva Slaninova, Diana Cernayova, Zuzana Sedrlova, Katerina Mrazova, Petr Sedlacek, Jana Nebesarova, Stanislav Obruca

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Cyanobacteria are gram-negative prokaryotes belonging to a group of photosynthetic bacteria. In comparison with heterotrophic microorganisms, cyanobacteria utilize atmospheric nitrogen and carbon dioxide without any additional substrates. This ability of these microorganisms could be employed in biotechnology for the production of bioplastics, concretely polyhydroxyalkanoates (PHAs) which are primarily accumulated as a storage material in cells in the form of intracellular granules. In this study, there two cyanobacterial cultures from genera Synechocystis were used, namely Synechocystic sp. PCC 6803 and Synechocystis salina CCALA 192. There were optimized and used several various approaches, including microscopic techniques such as cryo-scanning electron microscopy (Cryo-SEM) and transmission electron microscopy (TEM), and fluorescence lifetime imaging microscopy using Nile red as a fluorescent probe (FLIM). Due to these instrumental techniques, the morphology of intracellular space and surface of cells were characterized. The next group of methods which were employed was spectroscopic techniques such as UV-Vis spectroscopy measured in two modes (turbidimetry and integration sphere) and Fourier transform infrared spectroscopy (FTIR). All these diverse techniques were used for the detection and characterization of pigments (chlorophylls, carotenoids, phycocyanin, etc.) and PHAs, in our case poly (3-hydroxybutyrate) (P3HB). To verify results, gas chromatography (GC) was employed concretely for the determination of the amount of P3HB in biomass. Cyanobacteria were also characterized as polyhydroxybutyrate producers by flow cytometer, which could count cells and at the same time distinguish cells including P3HB and without due to fluorescent probe called BODIPY and live/dead fluorescent probe SYTO Blue. Based on results, P3HB content in cyanobacteria cells was determined, as also the overall fitness of the cells. Acknowledgment: Funding: This study was partly funded by the projectGA19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), project I 4082-B25.

Keywords: cyanobacteria, fluorescent probe, microscopic techniques, poly(3hydroxybutyrate), spectroscopy, chromatography

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Authors: Miroslav Hyza, Petr Rulik, Vojtech Bednar, Jan Sury

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An early and reliable detection of an increased radioactivity level in the atmosphere is one of the key aspects of atmospheric radiological monitoring. Although the standard laboratory procedures provide detection limits as low as few µBq/m³, their major drawback is the delayed result reporting: typically a few days. This issue is the main objective of the HAMRAD project, which gave rise to a prototype of an autonomous monitoring device. It is based on the idea of sequential aerosol sampling using a carrousel sample changer combined with a gamma-ray spectrometer. In our hardware configuration, the air is drawn through a filter positioned on the carrousel so that it could be rotated into the measuring position after a preset sampling interval. Filter analysis is performed via a 50% HPGe detector inside an 8.5cm lead shielding. The spectrometer output signal is then analyzed using DSP electronics and Gamwin software with preset nuclide libraries and other analysis parameters. After the counting, the filter is placed into a storage bin with a capacity of 250 filters so that the device can run autonomously for several months depending on the preset sampling frequency. The device is connected to a central server via GPRS/GSM where the user can view monitoring data including raw spectra and technological data describing the state of the device. All operating parameters can be remotely adjusted through a simple GUI. The flow rate is continuously adjustable up to 10 m³/h. The main challenge in spectrum analysis is the natural background subtraction. As detection limits are heavily influenced by the deposited activity of radon decay products and the measurement time is fixed, there must exist an optimal sample decay time (delayed spectrum acquisition). To solve this problem, we adopted a simple procedure based on sequential spectrum acquisition and optimal partial spectral sum with respect to the detection limits for a particular radionuclide. The prototyped device proved to be able to detect atmospheric contamination at the level of mBq/m³ per an 8h sampling.

Keywords: aerosols, atmosphere, atmospheric radioactivity monitoring, autonomous sampler

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Authors: Martin Tazky, Rudolf Hela, Lucia Osuska, Petr Novosad

Abstract:

Concrete’s volumetric changes are natural process caused by silicate minerals’ hydration. These changes can lead to cracking and subsequent destruction of cementitious material’s matrix. In most cases, cracks can be assessed as a negative effect of hydration, and in all cases, they lead to an acceleration of degradation processes. Preventing the formation of these cracks is, therefore, the main effort. Once of the possibility how to eliminate this natural concrete shrinkage process is by using different types of dispersed reinforcement. For this application of concrete shrinking, steel and polymer reinforcement are preferably used. Despite ordinarily used reinforcement in concrete to eliminate shrinkage it is possible to look at this specific problematic from the beginning by itself concrete mix composition. There are many secondary raw materials, which are helpful in reduction of hydration heat and also with shrinkage of concrete during curing. The new science shows the possibilities of shrinkage reduction also by the controlled formation of hydration products, which could act by itself morphology as a traditionally used dispersed reinforcement. This contribution deals with the possibility of controlled formation of mono- and tri-sulfate which are considered like degradation minerals. Mono- and tri- sulfate's controlled formation in a cementitious composite can be classified as a self-healing ability. Its crystal’s growth acts directly against the shrinking tension – this reduces the risk of cracks development. Controlled formation means that these crystals start to grow in the fresh state of the material (e.g. concrete) but stop right before it could cause any damage to the hardened material. Waste materials with the suitable chemical composition are very attractive precursors because of their added value in the form of landscape pollution’s reduction and, of course, low cost. In this experiment, the possibilities of using the fly ash from fluidized bed combustion as a mono- and tri-sulphate formation additive were investigated. The experiment itself was conducted on cement paste and concrete and specimens were subjected to a thorough analysis of physicomechanical properties as well as microstructure from the moment of mixing up to 180 days. In cement composites, were monitored the process of hydration and shrinkage. In a mixture with the used admixture of fluidized bed combustion fly ash, possible failures were specified by electronic microscopy and dynamic modulus of elasticity. The results of experiments show the possibility of shrinkage concrete reduction without using traditionally dispersed reinforcement.

Keywords: shrinkage, monosulphates, trisulphates, self-healing, fluidized fly ash

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Authors: Eiman A. Al-Enezi, Gin Jose, Sikha Saha, Paul Millner

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Rare earth nanotechnology has gained a considerable amount of interest in the field of biosensing due to the unique luminescence properties of lanthanides. Chelating rare earth ions plays a significant role in biological labelling applications including medical diagnostics, due to their different excitation and emission wavelengths, variety of their spectral properties, sharp emission peaks and long fluorescence lifetimes. We aimed to develop a platform for biosensors based on Europium (Eu³⁺) chelates against biomarkers of cardiac injury (heart-type fatty acid binding protein; H-FABP3) and stroke (glial fibrillary acidic protein; GFAP). Additional novelty in this project is the use of synthetic binding proteins (Affimers), which could offer an excellent alternative targeting strategy to the existing antibodies. Anti-GFAP and anti-HFABP3 Affimer binders were modified to increase the number of carboxy functionalities. Europium nitrate then incubated with the modified Affimer. The luminescence characteristics of the Eu³⁺ complex with modified Affimers and antibodies against anti-GFAP and anti-HFABP3 were measured against different concentrations of the respective analytes on excitation wavelength of 395nm. Bovine serum albumin (BSA) was used as a control against the IgG/Affimer Eu³⁺ complexes. The emission spectrum of Eu³⁺ complex resulted in 5 emission peaks ranging between 550-750 nm with the highest intensity peaks were at 592 and 698 nm. The fluorescence intensity of Eu³⁺ chelates with the modified Affimer or antibodies increased significantly by 4-7 folder compared to the emission spectrum of Eu³⁺ complex. The fluorescence intensity of the Affimer complex was quenched proportionally with increased analyte concentration, but this did not occur with antibody complex. In contrast, the fluorescence intensity for Eu³⁺ complex increased slightly against increased concentration of BSA. These data demonstrate that modified Affimers Eu³⁺ complexes can function as nanobiosensors with potential diagnostic and analytical applications.

Keywords: lanthanides, europium, chelates, biosensors

Procedia PDF Downloads 493