Search results for: markov chain mote carlo

Authors: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

Procedia PDF Downloads 125

Authors: Elif Tugce Aksun Tumerkan, Chris Lowe, Hannah Krupa

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Fluorescent proteins are self-sufficient in forming chromophores with a visible wavelength from 3 amino acids sequence within their own polypeptide structure. This chromophore – a molecule that absorbs a photon of light and exhibits an energy transition equal to the energy of the absorbed photon. Fluorescent proteins (FPs) consisted of a chain of 238 amino acid residues and composed of 11 beta strands shaped in a cylinder surrounding an alpha helix structure. A better understanding of the system of the chromospheres and the increasing advance in protein engineering in recent years, the properties of FPs offers the potential for new applications. They have used sensors and probes in molecular biology and cell-based research that giving a chance to observe these FPs tagged cell localization, structural variation and movement. For clarifying functional uses of fluorescent proteins, electrophoretic properties of these proteins are one of the most important parameters. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is used for determining electrophoretic properties commonly. While there are many techniques are used for determining the functionality of protein-based research, SDS-PAGE analysis can only provide a molecular level assessment of the proteolytic fragments. Before SDS-PAGE analysis, fluorescent proteins need to successfully purified. Due to directly purification of the target, FPs is difficult from the animal, gene expression is commonly used which must be done by transformation with the plasmid. Furthermore, used gel within electrophoresis and staining agents properties have a key role. In this review, the different factors that have the impact on the electrophoretic properties of fluorescent proteins explored. Fluorescent protein separation and purification are the essential steps before electrophoresis that should be done very carefully. For protein purification, gene expression process and following steps have a significant function. For successful gene expression, the properties of selected bacteria for expression, used plasmid are essential. Each bacteria has own characteristics which are very sensitive to gene expression, also used procedure is the important factor for fluorescent protein expression. Another important factors are gel formula and used staining agents. Gel formula has an effect on the specific proteins mobilization and staining with correct agents is a key step for visualization of electrophoretic bands of protein. Visuality of proteins can be changed depending on staining reagents. Apparently, this review has emphasized that gene expression and purification have a stronger effect than electrophoresis protocol and staining agents.

Keywords: cell biology, gene expression, staining agents, SDS-page

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Authors: Oluwaseun Adeleke, Samuel O. Ikani, Fidelis Edet, Anthony Nwala, Mopelola Raji, Simeon Christian Chukwu

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Background: The Delivering Innovations in Selfcare (DISC) project aims to increase access to self-care options for women of reproductive age, starting with self-inject subcutaneous depot medroxyprogesterone acetate (DMPA-SC) contraception services. However, the project has faced challenges in ensuring the continuous availability of the commodity in health facilities. Although most states in the country rely on the federal ministry of Health for supplies, some are gradually funding the procurement of Family Planning (FP) commodities. This attempt is, however, often accompanied by procurement delays and purchases inadequate to meet demand. This dilemma was further exacerbated by the commencement of demand generation activities by the project in supported states which geometrically increased commodity utilization rates and resulted in receding stock and occasional service disruptions. Strategies: The project deployed various strategies were implemented to ensure the continuous availability of commodities. These include facilitating inter-facility transfer, monthly tracking of commodity utilization, and alerting relevant authorities when stock levels reach a minimum. And supporting state-level procurement of DMPA-SC commodities through catalytic interventions. Results: Effective monitoring of commodity inventory at the facility level and strategic engagement with federal and state-level logistics units have proven successful in mitigating stock-out of commodities. It has helped secure up to 13,000 units of DMPA-SC commodities from federal logistics units and enabled state units to prioritize supported sites. This has ensured the continuity of DMPA-SC services and an increasing trend in the practice of self-injection. Conclusion: A functional supply chain is crucial to achieving commodity security, and without it, health programs cannot succeed. Stakeholder engagement, stock management and catalytic interventions have provided both short- and long-term measures to mitigate stock-outs and ensured a consistent supply of commodities to clients.

Keywords: family planning, contraception, DMPA-SC, self-care, self-injection, commodities, stock-out

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Authors: Ertuğrul Yılmaz

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One of the most important occupational groups in the food chain from farm to fork is a veterinary medicine. This occupational group, which has important duties in the prevention of many zoonotic diseases and in public health, takes place in many critical control points from soil to our kitchen. It has important duties from mycotoxins transmitted from the soil to slaughterhouses or milk processing facilities. Starting from the soil, which constitutes 70% of mycotoxin contamination, up to the TMR made from raw materials obtained from the soil, there are all critical control points from feeding to slaughterhouses and milk production enterprises. We can take the precaution of mycotoxins such as Aflatoxin B1, Ochratoxin, Zearalenone, and Fumonisin, which we encounter on farms while in the field. It has been reported that aflatoxin B1 is a casenerogen and passes into milk in studies. It is likely that many mycotoxins pose significant threats to public health and will turn out to be even more dangerous over time. Even raw material storage and TMR preparation are very important for public health. The danger of fumonisin accumulating in the liver will be understood over time. Zoonotic diseases are also explained with examples. In this study, how important veterinarians are in terms of public health is explained with examples. In the two-year mycotoxin screenings, fumonisin mycotoxin was found to be very high in corn and corn by-products, and it was determined that it accumulated in the liver for a long time and remained cornic in animals. It has been determined that mycotoxins are present in all livestock feeds, poultry feeds, and raw materials, not alone, but in double-triple form. Starting from the end, mycotoxin scans should be carried out from feed to raw materials and from raw materials to soil. In this way, we prevent the transmission of mycotoxins to animals and from animals to humans. Liver protectors such as toxin binders, beta-glucan, mannan oligosaccharides, activated carbon, prebiotics, and silymarin were used in certain proportions in the total mixed ratio, and positive results were obtained. Humidity and temperature controls of raw material silos were made at certain intervals. Necropsy was performed on animals that died as a result of mycotoxicosis, and macroscopic photographs were taken of the organs. We have determined that the mycotoxin screening in experimental animals and the feeds made without detecting the presence and amount of bacterial factors affect the results of the project to be made. For this, a series of precautionary plans have been created, starting from the production processes.

Keywords: mycotoxins, feed safety, processes, public health

Procedia PDF Downloads 87

Authors: Natalia Uribe Corrales, María Fernanda Benavides Erazo, Santiago Henao Villegas

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Introduction: Transport is a crucial link in the porcine chain because it is considered a stressful event in the animal, due to it is a new environment, which generates new interactions, together with factors such as speed, noise, temperature changes, vibrations, deprivation of food and water. Therefore, inadequate handling at this stage can lead to bruises, musculoskeletal injuries, fatigue, and mortality, resulting in canal seizures and economic losses. Objective: To characterize the transport and driving practices for the mobilization of standing pigs directed to slaughter plants in the Valle de Aburrá, Antioquia, Colombia in 2017. Methods: A descriptive cross-sectional study was carried out with the transporters arriving at the slaughterhouses approved by National Institute for Food and Medicine Surveillance (INVIMA) during 2017 in the Valle de Aburrá. The process of obtaining the samples was made from probabilistic sampling. Variables such as journey time, mechanical technical certificate, training in animal welfare, driving speed, material, and condition of floors and separators, supervision of animals during the trip, load density and mortality were analyzed. It was approved by the ethics committee for the use and care of animals CICUA of CES University, Act number 14 of 2015. Results: 190 trucks were analyzed, finding that 12.4% did not have updated mechanical technical certificate; the transporters experience in pig’s transportation was an average of 9.4 years (d.e.7.5). The 85.8% reported not having received training in animal welfare. Other results were that the average speed was 63.04km/hr (d.e 13.46) and the 62% had floors in good condition; nevertheless, the 48% had bad conditions on separators. On the other hand, the 88% did not supervise their animals during the journey, although the 62.2% had an adequate loading density, in relation to the average mortality was 0.2 deaths/travel (d.e. 0.5). Conclusions: Trainers should be encouraged on issues such as proper maintenance of vehicles, animal welfare, obligatory review of animals during mobilization and speed of driving, as these poorly managed indicators generate stress in animals, increasing generation of injuries as well as possible accidents; also, it is necessary to continue to improve aspects such as aluminum floors and separators that favor easy cleaning and maintenance, as well as the appropriate handling in the density of load that generates animal welfare.

Keywords: animal welfare, driving practices, pigs, truck infrastructure

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Authors: Nurdan Olguncelik Kaplan, Aysen Akay

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Nowadays soil pollution has become a global problem. External added polluters to the soil are destroying and changing the structure of the soil and the problems are becoming more complex and in this sense the correction of these problems is going to be harder and more costly. Cadmium has got a fast mobility in the soil and plant system because of that cadmium can interfere very easily to the human and animal food chain and in the same time this can be very dangerous. The cadmium which is absorbed and stored by the plants is causing to many metabolic changes of the plants like; protein synthesis, nitrogen and carbohydrate metabolism, enzyme (nitrate reductase) activation, photo and chlorophyll synthesis. The biological function of cadmium is not known over the plants and it is not a necessary element. The plant is generally taking in small amounts the cadmium and this element is competing with the zinc. Cadmium is causing root damages. Buckwheat (Fagopyrum esculentum) is an important nutraceutical because of its high content of flavonoids, minerals and vitamins, and their nutritionally balanced amino-acid composition. Buckwheat has relatively high biomass productivity, is adapted to many areas of the world, and can flourish in sterile fields; therefore buckwheat plants are widely used for the phytoremediation process.The aim of this study were to evaluate the phytoremediation capacity of the high-yielding plant Buckwheat (Fagopyrum esculentum) in soils contaminated with Cd and Zn. The soils were applied to differrent doses cd(0-12.5-25-50-100 mg Cd kg−1 soil in the form of 3CdSO4.8H2O ) and Zn (0-10-30 mg Zn kg−1 soil in the form of ZnSO4.7H2O) and incubated about 60 days. Later buckwheat seeds were sown and grown for three mounth under greenhouse conditions. The test plants were irrigated by using pure water after the planting process. Buckwheat seeds (Gunes and Aktas species) were taken from Bahri Dagdas International Agricultural Research. After harvest, Cd and Zn concentrations of plant biomass and grain, yield and translocation factors (TFs) for Cd and Cd were determined. Cadmium accumulation in biomass and grain significantly increased in dose-dependent manner. Long term field trials are required to further investigate the potential of buckwheat to reclaimed the soil. But this could be undertaken in conjunction with actual remediation schemes. However, the differences in element accumulation among the genotypes were affected more by the properties of genotypes than by the soil properties. Gunes genotype accumulated higher lead than Aktas genotypes.

Keywords: buckwheat, cadmium, phytoremediation, zinc

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Authors: Nishanth Venugopal Menon, Chuah Yon Jin, Samantha Phey, Wu Yingnan, Zhang Ying, Vincent Chan, Kang Yuejun

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Cell Migration is a vital phenomenon that the cells undergo in various physiological processes like wound healing, disease progression, embryogenesis, etc. Cell migration depends primarily on the chemical and physical cues available in the cellular environment. The chemical cue involves the chemokines secreted and gradients generated in the environment while physical cues indicate the impact of matrix properties like nanotopography and stiffness on the cells. Mesenchymal Stem Cells (MSCs) have been shown to have a role wound healing in vivo and its migration to the site of the wound has been shown to have a therapeutic effect. In the field of stem cell based tissue regeneration of bones and cartilage, one approach has been to introduce scaffold laden with MSCs into the site of injury to enable tissue regeneration. In this work, we have studied the combinatorial impact of the substrate physical properties on MSC migration. A microfluidic in vitro model was created to perform the migration studies. The microfluidic model used is a three compartment device consisting of two cell seeding compartments and one migration compartment. Four different PDMS substrates with varying substrate roughness, stiffness and hydrophobicity were created. Its surface roughness and stiffness was measured using Atomic Force Microscopy (AFM) while its hydrphobicity was measured from the water contact angle using an optical tensiometer. These PDMS substrates are sealed to the microfluidic chip following which the MSCs are seeded and the cell migration is studied over the period of a week. Cell migration was quantified using fluorescence imaging of the cytoskeleton (F-actin) to find out the area covered by the cells inside the migration compartment. The impact of adhesion proteins on cell migration was also quantified using a real-time polymerase chain reaction (qRT PCR). These results suggested that the optimal substrate for cell migration would be one with an intermediate level of roughness, stiffness and hydrophobicity. A higher or lower value of these properties affected cell migration negatively. These observations have helped us in understanding that different substrate properties need to be considered in tandem, especially while designing scaffolds for tissue regeneration as cell migration is normally impacted by the combinatorial impact of the matrix. These observations may lead us to scaffold optimization in future tissue regeneration applications.

Keywords: cell migration, microfluidics, in vitro model, stem cell migration, scaffold, substrate properties

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Authors: Yufeng Wu, Yifan Gu, Bin Li, Wei Wang

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Resource recycling process contains a subsystem with interactions of three dimensions including coupling allocation of primary and secondary resources, responsibility coordination of stakeholders in forward and reverse supply chains, and trans-boundary transfer of hidden resource and environmental responsibilities between regions. Overlap or lack of responsibilities is easy to appear at the intersection of the three management dimensions. It is urgent to make an overall design of the policy system for recycling resources. From theoretical perspective, this paper analyzes the unique external differences of resource and environment in various dimensions and explores the reason why the effects of trans-dimensional policies are strongly correlated. Taking the example of the copper resources contained in the waste electrical and electronic equipment, this paper constructs reduction effect accounting model of resources recycling and set four trans-dimensional policy scenarios including resources tax and environmental tax reform of the raw and secondary resources, application of extended producer responsibility system, promotion of clean development mechanism, and strict entry barriers of imported wastes. In these ways, the paper simulates the impact effect of resources recycling process on resource deduction and emission reduction of waste water and gas, and constructs trans-dimensional policy mix scenario through integrating dominant strategy. The results show that combined application of various dimensional policies can achieve incentive compatibility and the trans-dimensional policy mix scenario can reach a better effect. Compared with baseline scenario, this scenario will increase 91.06% copper resources reduction effect and improve emission reduction of waste water and gas by eight times from 2010 to 2030. This paper further analyzes the development orientation of policies in various dimension. In resource dimension, the combined application of compulsory, market and authentication methods should be promoted to improve the use ratio of secondary resources. In supply chain dimension, resource value, residual functional value and potential information value contained in waste products should be fully excavated to construct a circular business system. In regional dimension, it should give full play to the comparative advantages of manufacturing power to improve China’s voice in resource recycling in the world.

Keywords: resource recycling, trans-dimension, policy design, incentive compatibility, life cycle

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Authors: Tanima Chatterjee, Maitree Bhattacharyya

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The global challenge of type 2 diabetes mellitus is a major health concern in this millennium, and researchers are continuously exploring new targets to develop a novel therapeutic strategy. Type 2 diabetes mellitus (T2DM) is often coupled with dyslipidemia increasing the risks for cardiovascular (CVD) complications. Enhanced oxidative and nitrosative stresses appear to be the major risk factors underlying insulin resistance, dyslipidemia, β-cell dysfunction, and T2DM pathogenesis. Autophagy emerges to be a promising defense mechanism against stress-mediated cell damage regulating tissue homeostasis, cellular quality control, and energy production, promoting cell survival. In this study, we have attempted to explore the pivotal role of autophagy in T2DM subjects with or without dyslipidemia in peripheral blood mononuclear cells and insulin-resistant HepG2 cells utilizing flow cytometric platform, confocal microscopy, and molecular biology techniques like western blotting, immunofluorescence, and real-time polymerase chain reaction. In the case of T2DM with dyslipidemia higher population of autophagy, positive cells were detected compared to patients with the only T2DM, which might have resulted due to higher stress. Autophagy was observed to be triggered both by oxidative and nitrosative stress revealing a novel finding of our research. LC3 puncta was observed in peripheral blood mononuclear cells and periphery of HepG2 cells in the case of the diabetic and diabetic-dyslipidemic conditions. Increased expression of ATG5, LC3B, and Beclin supports the autophagic pathway in both PBMC and insulin-resistant Hep G2 cells. Upon blocking autophagy by 3-methyl adenine (3MA), the apoptotic cell population increased significantly, as observed by caspase‐3 cleavage and reduced expression of Bcl2. Autophagy has also been evidenced to control oxidative stress-mediated up-regulation of inflammatory markers like IL-6 and TNF-α. To conclude, this study elucidates autophagy to play a protective role in the case of diabetes mellitus with dyslipidemia. In the present scenario, this study demands to have a significant impact on developing a new therapeutic strategy for diabetic dyslipidemic subjects by enhancing autophagic activity.

Keywords: autophagy, apoptosis, dyslipidemia, reactive oxygen species, reactive nitrogen species, Type 2 diabetes

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Authors: Narendra Akhadkar, Philippe Delcambre

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In power protection and control products, assembly process variations due to the individual parts manufactured from single or multi-cavity tooling is a major problem. The dimensional and geometrical variations on the individual parts, in the form of manufacturing tolerances and assembly tolerances, are sources of clearance in the kinematic joints, polarization effect in the joints, and tolerance stack-up. All these variations adversely affect the quality of product, functionality, cost, and time-to-market. Variation simulation analysis may be used in the early product design stage to predict such uncertainties. Usually, variations exist in both manufacturing processes and materials. In the tolerance analysis, the effect of the dimensional and geometrical variations of the individual parts on the functional characteristics (conditions) of the final assembled products are studied. A functional characteristic of the product may be affected by a set of interrelated dimensions (functional parameters) that usually form a geometrical closure in a 3D chain. In power protection and control products, the prerequisite is: when a fault occurs in the electrical network, the product must respond quickly to react and break the circuit to clear the fault. Usually, the response time is in milliseconds. Any failure in clearing the fault may result in severe damage to the equipment or network, and human safety is at stake. In this article, we have investigated two important functional characteristics that are associated with the robust performance of the product. It is demonstrated that the experimental data obtained at the Schneider Electric Laboratory prove the very good prediction capabilities of the variation simulation performed using CETOL (tolerance analysis software) in an industrial context. Especially, this study allows design engineers to better understand the critical parts in the product that needs to be manufactured with good, capable tolerances. On the contrary, some parts are not critical for the functional characteristics (conditions) of the product and may lead to some reduction of the manufacturing cost, ensuring robust performance. The capable tolerancing is one of the most important aspects in product and manufacturing process design. In the case of miniature circuit breaker (MCB), the product's quality and its robustness are mainly impacted by two aspects: (1) allocation of design tolerances between the components of a mechanical assembly and (2) manufacturing tolerances in the intermediate machining steps of component fabrication.

Keywords: geometrical variation, product robustness, tolerance analysis, variation simulation

Procedia PDF Downloads 164

Authors: Taus Muganda

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This research explores the intricate relationship between e-procurement adoption and effective service delivery in the Uganda Coffee Industry, focusing on the processes involved, key actors, and the impact of digital transformation. The study is guided by three prominent theories, Actor-Network Theory, Resource-Based View Theory, and Institutional Theory to comprehensively explore the dynamics of e-procurement in the context of the coffee sector. The primary aim of this project is to examine the e-procurement adoption process and its role in enhancing service delivery within the Uganda Coffee Industry. The research questions guiding this inquiry are: firstly, whether e-procurement adoption and implementation contribute to achieving quality service delivery; and secondly, how e-procurement adoption can be effectively realized within the Uganda Coffee Industry. To address these questions, the study has laid out specific objectives. Firstly, it seeks to investigate the impact of e-procurement on effective service delivery, analysing how the integration of digital processes influences the overall quality of services provided in the coffee industry. Secondly, it aims to critically analyse the measures required to achieve effective delivery outcomes through the adoption and implementation of e-procurement, assessing the strategies that can maximize the benefits of digital transformation. Furthermore, the research endeavours to identify and examine the key actor’s instrumental in achieving effective service delivery within the Uganda Coffee Industry. By utilizing Actor-Network Theory, the study will elucidate the network of relationships and collaborations among actors involved in the e-procurement process. The research contributes to addressing a critical gap in the sector. Despite coffee being the leading export crop in Uganda, constituting 16% of total exports, there is a recognized need for digital transformation, specifically in the realm of e-procurement, to enhance the productivity of producers and contribute to the economic growth of the country. The study aims to provide insights into transforming the Uganda Coffee Industry by focusing on improving the e-procurement services delivered to actors in the coffee sector. The three forms of e-procurement investigated in this research—E-Sourcing, E-Payment, and E-Invoicing—serve as focal points in understanding the multifaceted dimensions of digital integration within the Uganda Coffee Industry. This research endeavours to offer practical recommendations for policymakers, industry stakeholders, and the UCDA to strategically leverage e-procurement for the benefit of the entire coffee value chain.

Keywords: e-procurement, effective service delivery, actors, actor-network theory, resource-based view theory, institutional theory, e-invocing, e-payment, e-sourcing

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Authors: Devi Eka Septiyani Arifin, Jrjeng Ruan

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As an approach to manipulate the performance of polymer thin film, epitaxy crystallization within polymer blends of poly(vinylidene fluoride) (PVDF) and its copolymer poly(vinylidene fluoride-trifluoroethylene) P(VDF-TrFE) was studied in this research, which involves the competition between phase separation and crystal growth of constitutive semicrystalline polymers. The unique piezoelectric feature of poly(vinylidene fluoride) crystalline phase is derived from the packing of molecular chains in all-trans conformation, which spatially arranges all the substituted fluorene atoms on one side of the molecular chain and hydrogen atoms on the other side. Therefore, the net dipole moment is induced across the lateral packing of molecular chains. Nevertheless, due to the mutual repulsion among fluorene atoms, this all-trans molecular conformation is not stable, and ready to change above curie temperature, where thermal energy is sufficient to cause segmental rotation. This research attempts to explore whether the epitaxial interactions between piezoelectric crystals and crystal lattice of hexamethylbenzene (HMB) crystalline platelet is able to stabilize this metastable all-trans molecular conformation or not. As an aromatic crystalline compound, the melt of HMB was surprisingly found able to dissolve the poly(vinylidene fluoride), resulting in homogeneous eutectic solution. Thus, after quenching this binary eutectic mixture to room temperature, subsequent heating or annealing processes were designed to explore the involve phase separation and crystallization behavior. The phase transition behaviors were observed in-situ by X-ray diffraction and differential scanning calorimetry (DSC). The molecular packing was observed via transmission electron microscope (TEM) and the principles of electron diffraction were brought to study the internal crystal structure epitaxially developed within thin films. Obtained results clearly indicated the occurrence of heteroepitaxy of PVDF/PVDF-TrFE on HMB crystalline platelet. Both the concentration of poly(vinylidene fluoride) and the mixing ratios of these two constitutive polymers have been adopted as the influential factors for studying the competition between the epitaxial crystallization of PVDF and P(VDF-TrFE) on HMB crystalline. Furthermore, the involved epitaxial relationship is to be deciphered and studied as a potential factor capable of guiding the wide spread of piezoelectric crystalline form.

Keywords: epitaxy, crystallization, crystalline platelet, thin film and mixing ratio

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Authors: A. B. Moldes, X. Vecino, L. Rodriguez-López, J. M. Dominguez, J. M. Cruz

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The concept of the circular economy is focus on the importance of providing goods and services sustainably. Thus, in a future it will be necessary to respond to the environmental contamination and to the use of renewables substrates by moving to a more restorative economic system that drives towards the utilization and revalorization of residues to obtain valuable products. During its evolution our industrial economy has hardly moved through one major characteristic, established in the early days of industrialization, based on a linear model of resource consumption. However, this industrial consumption system will not be maintained during long time. On the other hand, there are many industries, like the corn milling industry, that although does not consume high amount of non renewable substrates, they produce valuable streams that treated accurately, they could provide additional, economical and environmental, benefits by the extraction of interesting commercial renewable products, that can replace some of the substances obtained by chemical synthesis, using non renewable substrates. From this point of view, the use of streams from corn milling industry to obtain surface-active compounds will decrease the utilization of non-renewables sources for obtaining this kind of compounds, contributing to a circular and global economy. However, the success of the circular economy depends on the interest of the industrial sectors in the revalorization of their streams by developing relevant and new business models. Thus, it is necessary to invest in the research of new alternatives that reduce the consumption of non-renewable substrates. In this study is proposed the utilization of a corn milling industry stream to obtain an extract with surfactant capacity. Once the biosurfactant is extracted, the corn milling stream can be commercialized as nutritional media in biotechnological process or as animal feed supplement. Usually this stream is combined with other ingredients obtaining a product namely corn gluten feed or may be sold separately as a liquid protein source for beef and dairy feeding, or as a nutritional pellet binder. Following the productive scheme proposed in this work, the corn milling industry will obtain a biosurfactant extract that could be incorporated in its productive process replacing those chemical detergents, used in some point of its productive chain, or it could be commercialized as a new product of the corn manufacture. The biosurfactants obtained from corn milling industry could replace the chemical surfactants in many formulations, and uses, and it supposes an example of the potential that many industrial streams could offer for obtaining valuable products when they are manage properly.

Keywords: biosurfactantes, circular economy, corn, sustainability

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Authors: Miguel Llorens-Marin, Adolfo Hernandez, Maria Puelles-Gallo

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The increasing influence of Online Travel Agencies (OTAs) on hotel bookings and the electronic word-of-mouth (eWOM) contained in them has been featured by many scientific studies as a major factor in the booking decision. The main reason is that nowadays, in the hotel sector, consumers first come into contact with the offer through the web and the online environment. Due to the nature of the hotel product and the fact that it is booked in advance to actually seeing it, there is a lack of knowledge about its actual features. This makes eWOM a major channel to help consumers to reduce their perception of risk when making their booking decisions. This research studies the relationship between aspects of customer influenceability by reading eWOM communications, at the time of booking a hotel, with the propensity to write a review. In other words, to test relationships between the reading and the writing of eWOM. Also investigates the importance of different underlying motivations for writing eWOM. Online surveys were used to obtain the data from a sample of hotel customers, with 739 valid questionnaires. A measurement model and Path analysis were carried out to analyze the chain of relationships among the independent variable (influenceability from reading reviews) and the dependent variable (propensity to write a review) with the mediating effects of additional variables, which help to explain the relationship. The authors also tested the moderating effects of age and gender in the model. The study considered three different underlying motivations for writing a review on a hotel experience, namely hedonic, altruistic and conflicted. Results indicate that the level of influenceability by reading reviews has a positive effect on the propensity to write reviews; therefore, we manage to link the reading and the writing of reviews. Authors also discover that the main underlying motivation to write a hotel review is the altruistic motivation, being the one with the higher Standard regression coefficient above the hedonic motivation. The authors suggest that the propensity to write reviews is not related to sociodemographic factors (age and gender) but to attitudinal factors such as ‘the most influential factor when reading’ and ‘underlying motivations to write. This gives light on the customer engagement motivations to write reviews. The implications are that managers should encourage their customers to write eWOM reviews on altruistic grounds to help other customers to make a decision. The most important contribution of this work is to link the effect of reading hotel reviews with the propensity to write reviews.

Keywords: hotel reviews, electronic word-of-mouth (eWOM), online consumer reviews, digital marketing, social media

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Authors: Hasan Topacoglu

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The Meiji Restoration was a chain of events that restored Japan in 1868 and considered as the beginning of Japanese Modernization by many scholars. In 1968, to honor its modern incarnation, Japan celebrated Meiji Centennial as one of the biggest Media Events in the country after the World War II. It was celebrated all around the country throughout the year following with a central event in Tokyo. Meanwhile, Japanese scholars started an opposition movement and claimed that Government was using this event to raise nationalism, pointing at Government’s statement on the meaning of Meiji. Most of the scholars, unfortunately, were hooked into the ideological problem of the Government’s way of planning and evaluated it as a failure. However, scholars missed out an important point that apart from the central event in Tokyo, each city planned its own event and celebrated it on a different date, also with a different theme. For example, Kyoto showed a regional characteristic and focused on Kyoto’s own culture, tradition etc., and highlighted a further past than 100 years. This was mainly because some areas/cities had a different ‘memory’ for Meiji Restoration than Tokyo which was reflected through the way they celebrated Meiji Centennial. On the other hand, 2023 will be the year of Turkish Republic Centennial. A year which will be marked by national and maybe even international events. Although an official committee has not been announced yet, The 2023 Vision, a list of goals has been released by the Government to coincide with the centenary of the Republic of Turkey in 2023 and there are some ongoing projects that are planned to be completed by then. By looking at the content of these projects, it is possible to say that Government is aiming to focus on Modernization through the Centennial. However, some of the projects are already showing some interesting characteristics such as the Istanbul New Airport whose design is inspired by Selimiye Mosque’s Islamic-Ottoman figure. It is true that Turkey and Japan have different historical backgrounds and the timeline of the Meiji Restoration and Foundation of Turkish Republic are different. Therefore, a particular comparison between these two events is not justified. However, they may have more in common than we are up to think because, each country marked the start of a new nation conceived on modern principles. For that reason, it is important to understand the similarities or differences between Meiji Centennial and Turkish Republic Centennial as a media event. This study introduces Meiji Centennial as a media event and analyses opposition movement along with the meaning of Meiji Centennial. Additionally, it explains regional characteristic differences and gives Kyoto as an example. Moreover, it introduces some of the ongoing Centennial projects in Turkey and analyses the meaning of the Turkish Republic Centennial through these projects. Without comparing Japan and Turkey, it explains the case of Japan but the discussion centers on deepening our understanding of Centennial as a Media Event and remarks some important aspects for Turkey’s upcoming Centennial events.

Keywords: media events, Meiji centennial, the 2023 vision, Turkish republic centennial

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Authors: Leslie Nobler

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African fabrics based specifically on the Dutch Wax Print, or Ankara, popularized during Africa's colonial era, have had an enormous impact on western fashion (especially in the US and UK), in the last half-decade. The trend has had an effect on the world of visual arts as well, which circuitously, also heavily impacts fashion design. In fashion, and notably in celebrity apparel choices, this is in part due to ‘identity’ and taking pride in one's African roots; in the visual arts, artists such as Yinka Shonibare and Njideka Akunyili Crosby are making statements about identity politics, colonialism up through post-colonialism, and racism. The ‘global village’ brought on by the internet has driven this proliferation, as have improvements in the printing technology with which the Ankara print is made, combining wax-resist with roller printing. The newest patterns can now be designed authentically in western African and easily sent electronically to Europe for printing. Examples of Ankara's new reach across the Atlantic abound. They have taken several paths, which the paper will detail. Briefly, the first is its greater utilization in the fashion world, from authentic textile shops in African American neighborhoods to copied (knocked-off) low-end reproductions in discount chains. Secondly, we are seeing far more uses of these textiles/patterns in important works of fine arts from major museums, in Philadelphia to Palm Beach to the Mass MOCA (in the US), all the way to the Israel Museum in Jerusalem, and everywhere in between. And lastly, but quite significantly, we see this trend throughout social media thanks to Instagram, Pinterest and celebrity photos –even at the recent royal wedding. What shall sustain this major new design direction is that Ankara changes with and adapts to the times. Some of it is now printed in West Africa, often in the Nigeria area. And some may be designed in Europe or even at knock-off apparel studios in NY or Asia. But it stays utterly relevant because the motifs are based on objects and scenes in everyday life. In my design studio and university design classes, this idea is first and foremost, from our big spiritual eye motifs to drawings of our art supplies to the ‘politically-loaded’ chain patterns. This first-hand creativity experience becomes part of the research of this paper, along with historic and contemporary sources of inquiry, both through a literature/image search and anecdotal experience into what is behind this exciting and surprising trend.

Keywords: African wax print, Ankara, identity (politics), textile design, surface design

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Authors: Katarzyna Miśkiewicz, Magdalena Lasoń-Rydel, Małgorzata Krępska, Katarzyna Sieczyńska, Iwona Masłowska-Lipowicz, Katarzyna Ławińska

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The aim of the study was to analyse a selection of 12 pasteurised products made from fruit and vegetables, such as fruit juices, fruit drinks, jams, marmalades and jam produced by small regional factories as well as large food corporations. The research was carried out as part of the project "Innovative system of healthy and regional food distribution", funded by the Ministry of Education and Science (Poland), which aims to create an economically and organisationally strong agri-food industry in Poland through effective cooperation between scientific and socio-economic actors. The main activities of the project include support for the creation of new distribution channels for regional food products and their easy access to a wide group of potential customers while maintaining the highest quality standards. One of the key areas of the project is food quality analyses conducted to indicate the competitive advantage of regional products. Presented here are studies on the content of sugars, vitamin C, preservatives, synthetic colours, sweeteners, sodium and potassium, as well as studies on the microbiological purity of selected products made from fruit and vegetables. The composition of products made from fruit and vegetables varies greatly and depends on both the type of raw material and the way it is processed. Of the samples tested, fruit drinks contained the least amount of sugars, and jam and marmalade made by large producers and bought in large chain stores contained the most. However, the low sugar content of some fruit drinks is due to the presence of the sweetener sucralose in their composition. The vitamin C content of the samples varied, being higher in products where it was added during production. All products made in small local factories were free of food additives such as preservatives, sweeteners and synthetic colours, indicating their superiority over products made by large producers. Products made in small local factories were characterised by a relatively high potassium content. The microbiological purity of commercial products was confirmed - no Salmonella spp. were detected, and the number of mesophilic bacteria, moulds, yeasts, and β-glucuronidase-positive E. coli was below the limit of quantification.

Keywords: fruit and vegetable products, sugars, food additives, HPLC, ICP-OES

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Authors: Manisha, Manisha Mathur, Jay K. Desai, Shesh Asopa, Manisha Mehra

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The present study was carried out for the comprehensive analysis of lumpy skin disease (LSD) in cattle and to elucidate the histopathology of vital organs in natural outbreaks. Lumpy skin disease (LSD) is a viral infection that primarily affects cattle. It is caused by a Capri pox virus and is characterized by the formation of skin nodules or lesions. For this study, a postmortem of 20 cows who died of Lumpy skin disease in different regions of Rajasthan was conducted. This study aimed to examine a cow's external and internal organs to confirm if lumpy skin disease was the cause of death. Accurate diagnosis is essential for improving disease surveillance, understanding the disease's progression, and informing control measures. Pathological examinations reveal virus-induced changes across organs, while histopathological analyses provide crucial insights into the disease's pathogenesis, aiding in the development of advanced diagnostics and effective prevention strategies. Histopathological examination of nodular skin lesions revealed edema, hyperemia, acanthosis, severe hydropic degeneration/ballooning degeneration, and hyperkeratosis in the epidermis. In the lungs, congestion, oedema, emphysema, and atelectasis were observed grossly. Microscopically changes were suggestive of interstitial pneumonia, suppurative pneumonia, bronchopneumonia post pneumonic fibrosis, and stage of resolution. Grossely liver showed congestion and necrotic foci microscopically in most of the cases, and the liver showed acute viral hepatitis. Microscopically in kidneys, multifocal interstitial nephritis was observed. There was marked interstitial inflammation and zonal fibrosis with cystically dilated tubules and bowman's capsules. Microscopically, most of the heart tissue section showed normal histology with few sarcocysts in between cardiac muscles. In some cases, loss of cross striation, sarcoplasmic vacuolation, fregmentation, and disintegration of cardiac fibres were observed. The present study revealed the characteristic gross and histopathological changes in different organs in natural cases of lumpy skin disease. Further, the disease was confirmed based on the molecular diagnosis and transmission electron microscopy of capripox infection in the affected cattle in the study area.

Keywords: Capripoxvirus, lumpy skin disease, polymerage chain reaction, transmission electron microscopy

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Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

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Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

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Authors: Chiranjit Das, Sanjay Jharkharia

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This study is focused on transport mode selection under lead time variability and emissions constraint. In order to reduce the carbon emissions generation due to transportation, organization has often faced a dilemmatic choice of transport mode selection since logistic cost and emissions reduction are complementary with each other. Another important aspect of transportation decision is lead-time variability which is least considered in transport mode selection problem. Thus, in this study, we provide a comprehensive mathematical based analytical model to decide transport mode selection under emissions constraint. We also extend our work through analysing the effect of lead time variability in the transport mode selection by a sensitivity analysis. In order to account lead time variability into the model, two identically normally distributed random variables are incorporated in this study including unit lead time variability and lead time demand variability. Therefore, in this study, we are addressing following questions: How the decisions of transport mode selection will be affected by lead time variability? How lead time variability will impact on total supply chain cost under carbon emissions? To accomplish these objectives, a total transportation cost function is developed including unit purchasing cost, unit transportation cost, emissions cost, holding cost during lead time, and penalty cost for stock out due to lead time variability. A set of modes is available to transport each node, in this paper, we consider only four transport modes such as air, road, rail, and water. Transportation cost, distance, emissions level for each transport mode is considered as deterministic and static in this paper. Each mode is having different emissions level depending on the distance and product characteristics. Emissions cost is indirectly affected by the lead time variability if there is any switching of transport mode from lower emissions prone transport mode to higher emissions prone transport mode in order to reduce penalty cost. We provide a numerical analysis in order to study the effectiveness of the mathematical model. We found that chances of stock out during lead time will be higher due to the higher variability of lead time and lad time demand. Numerical results show that penalty cost of air transport mode is negative that means chances of stock out zero, but, having higher holding and emissions cost. Therefore, air transport mode is only selected when there is any emergency order to reduce penalty cost, otherwise, rail and road transport is the most preferred mode of transportation. Thus, this paper is contributing to the literature by a novel approach to decide transport mode under emissions cost and lead time variability. This model can be extended by studying the effect of lead time variability under some other strategic transportation issues such as modal split option, full truck load strategy, and demand consolidation strategy etc.

Keywords: carbon emissions, inventory theoretic model, lead time variability, transport mode selection

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Authors: Joao Ferreira Do Rosario, Nuno Gustavo, Ana Machado, Lurdes Calisto, Luisa Carvalho, Georgette Andraz

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Currently, tourism small and medium enterprises (TSMEs) face serious economic and financial problems, making recovery efforts difficult. How the pandemic will affect tourists' behavior is still to be known. Will tourists be even more cautious regarding their choices or, on the contrary, will they be more adventurers with an enormous desire to travel in search of the lost freedom? Tourists may become even more demanding when traveling, more austere, or less concerned and eager to socialize. Adjusting to this "new tourist" is an added challenge for tourism service providers. Generation Z made up of individuals born in 1995 and following years, currently tends to assume a particular role and meaning in the present and future economic and social context, considering that we are facing the youngest workforce as well as tomorrow's consumers. This generation is distinguished from others as it is the first generation to combine a high level of education and technological knowledge and to fully experience the digital world. These young people are framed by a new value system that can explain new behaviours and consumption, namely, in the context of tourism. All these considerations point to the importance of investigating this target group as it is essential to understand how these individuals perceive, understand, act, and can be involved in a new environment built around a society regulated by new priorities and challenges of a sustainable nature. This leads not only to a focus on short-term market choices but mainly to predict future choices from a longer-term perspective. Together with the social background of a person, values are considered a stable antecedent of behavior and might therefore predict not just immediate, but also future choices. Furthermore, the meaning attributed to travel has a general connotation and goes beyond a specific travel choice or experience. In other words, values and travel's meaning form a chain of influences on the present and future travel behavior. This study explores the social background and values of Generation Z travelers vs the meaning these tourists give to travel. The aim is to discover in their present behavior cues to predict travel choices so that the future of tourism can be secured. This study also provides data for predicting the tourism choices of youngsters in the more immediate future. Methodologically, a quantitative approach was adopted based on the collection of data through a survey. Since academic research on Generation Z of tourists is still scarce, it is expected to contribute to deepening scientific knowledge in this area. Furthermore, it is expected that this research will support tourism professionals in defining differentiated marketing strategies and adapted to the requirements of this target, in a new time.

Keywords: Generation Z, travel behavior, travel meaning, Generation Z Values

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Authors: Sibashish Baksi, Ujjaini Sarkar, Sudeshna Saha

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In the present study, hydrolysis of Pinus roxburghi wood powder was carried out with Viscozyme, and kinetics of the hydrolysis has been investigated. Finely ground sawdust is submerged into 2% aqueous peroxide solution (pH=11.5) and pretreated through autoclaving, probe sonication, and alkaline peroxide pretreatment. Afterward, the pretreated material is subjected to hydrolysis. A chain of experiments was executed with delignified biomass (50 g/l) and varying enzyme concentrations (24.2–60.5 g/l). In the present study, 14.32 g/l of glucose, along with 7.35 g/l of xylose, have been recovered with a viscozyme concentration of 48.8 g/l and the same condition was treated as optimum condition. Additionally, thermal deactivation of viscozyme has been investigated and found to be gradually decreasing with escalated enzyme loading from 48.4 g/l (dissociation constant= 0.05 h⁻¹) to 60.5 g/l (dissociation constant= 0.02 h⁻¹). The hydrolysis reaction is a pseudo first-order reaction, and therefore, the rate of the hydrolysis can be expressed as a fractal-like kinetic equation that communicates between the product concentration and hydrolytic time t. It is seen that the value of rate constant (K) increases from 0.008 to 0.017 with augmented enzyme concentration from 24.2 g/l to 60.5 g/l. Greater value of K is associated with stronger enzyme binding capacity of the substrate mass. However, escalated concentration of supplied enzyme ensures improved interaction with more substrate molecules resulting in an enhanced de-polymerization of the polymeric sugar chains per unit time which eventually modifies the physiochemical structure of biomass. All fractal dimensions are in between 0 and 1. Lower the value of fractal dimension, more easily the biomass get hydrolyzed. It can be seen that with increased enzyme concentration from 24.2 g/l to 48.4 g/l, the values of fractal dimension go down from 0.1 to 0.044. This indicates that the presence of more enzyme molecules can more easily hydrolyze the substrate. However, an increased value has been observed with a further increment of enzyme concentration to 60.5g/l because of diffusional limitation. It is evident that the hydrolysis reaction system is a heterogeneous organization, and the product formation rate depends strongly on the enzyme diffusion resistances caused by the rate-limiting structures of the substrate-enzyme complex. Value of the rate constant increases from 1.061 to 2.610 with escalated enzyme concentration from 24.2 to 48.4 g/l. As the rate constant is proportional to Fick’s diffusion coefficient, it can be assumed that with a higher concentration of enzyme, a larger amount of enzyme mass dM diffuses into the substrate through the surface dF per unit time dt. Therefore, a higher rate constant value is associated with a faster diffusion of enzyme into the substrate. Regression analysis of time curves with various enzyme concentrations shows that diffusion resistant constant increases from 0.3 to 0.51 for the first two enzyme concentrations and again decreases with enzyme concentration of 60.5 g/l. During diffusion in a differential scale, the enzyme also experiences a greater resistance during diffusion of larger dM through dF in dt.

Keywords: viscozyme, glucose, fractal kinetics, thermal deactivation

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Authors: Ratneev Kaur, Tajinder Kaur, Anupam Kaur

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Introduction: Polycystic Ovary Syndrome (PCOS) is a heterogenous disorder of endocrine system among women of reproductive age. PCOS is characterized by hyperandrogenism, anovulation, polycystic ovaries, hirsutism, obesity, and hyperinsulinemia. Several pathways are implicated in its etiology including the metabolic pathway of steroid hormone synthesis regulatory pathways. PCOS is an androgen excess disorder, genes operating in steroidogenesis may alter pathogenesis of PCOS. The cytochrome P450scc is a cholesterol side chain cleavage enzyme coded by CYP11A1 gene and catalyzes conversion of cholesterol to pregnenolone, the initial and rate-limiting step in steroid hormone synthesis. It is postulated that polymorphisms in this gene may play an important role in the regulation of CYP11A1 expression and leading to increased or decreased androgen production. The present study will be the first study from north India to best of our knowledge, to analyse the association of CYP11A1 (rs11632698) polymorphism in women suffering from PCOS. Methodology: The present study was approved by ethical committee of Guru Nanak Dev University in consistent with declaration of Helsinki. A total of 300 samples (150 PCOS cases and 150 controls) were recruited from Hartej hospital, for the present study. Venous blood sample (3ml) was withdrawn from women diagnosed with PCOS by doctor, according to Rotterdam 2003 criteria and from healthy age matched controls only after informed consent and detailed filled proforma. For molecular genetics analysis, blood was stored in EDTA vials. After DNA isolation by organic method, PCR-RFLP approach was used for genotyping and association analysis of rs11632698 polymorphism. Statistical analysis was done to check for significance of selected polymorphism with PCOS. Results: In 150 PCOS cases, the frequency of AA, AG and GG genotype was found to be 48%, 35%, and 13% compared to 62%, 27% and 8% in 150 controls. The major allele (A) and minor allele (G) frequency was 68% and 32% in cases and 78% and 22% in controls. Minor allele frequency was higher in cases as compared to controls, as well as the distribution of genotype was observed to be statistically significant (ᵡ²=6.525, p=0.038). Odds ratio in dominant, co-dominant and recessive models observed was 1.81 (p=0.013), 1.54 (p=0.012) and 1.77 (p=0.132) respectively. Conclusion: The present study showed statistically significant association of rs11632698 with PCOS (p=0.038) in North Indian women.

Keywords: polycystic ovary syndrome, CYP11A1, rs11632698, hyperandrogenism

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Authors: Agnieszka H. Ludwig-Slomczynska, Michal T. Seweryn, Przemyslaw Kapusta, Ewelina Pitera, Katarzyna Cyganek, Urszula Mantaj, Lucja Dobrucka, Ewa Wender-Ozegowska, Maciej T. Malecki, Pawel Wolkow

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Obesity results from an imbalance between energy intake and its expenditure. Genome-Wide Association Study (GWAS) analyses have led to discovery of only about 100 variants influencing body mass index (BMI), which explain only a small portion of genetic variability. Analysis of gene epistasis gives a chance to discover another part. Since it was shown that interaction and communication between nuclear and mitochondrial genome are indispensable for normal cell function, we have looked for epistatic interactions between the two genomes to find their correlation with BMI. Methods: The analysis was performed on 366 T1DM patients using Illumina Infinium OmniExpressExome-8 chip and followed by imputation on Michigan Imputation Server. Only genes which influence mitochondrial functioning (listed in Human MitoCarta 2.0) were included in the analysis – variants of nuclear origin (MAF > 5%) in 1140 genes and 42 mitochondrial variants (MAF > 1%). Gene expression analysis was performed on GTex data. Association analysis between genetic variants and BMI was performed with the use of Linear Mixed Models as implemented in the package 'GENESIS' in R. Analysis of association between mRNA expression and BMI was performed with the use of linear models and standard significance tests in R. Results: Among variants involved in epistasis between mitochondria and nucleus we have identified one in mitochondrial transcription factor, TFB2M (rs6701836). It interacted with mitochondrial variants localized to MT-RNR1 (p=0.0004, MAF=15%), MT-ND2 (p=0.07, MAF=5%) and MT-ND4 (p=0.01, MAF=1.1%). Analysis of the interaction between nuclear variant rs6701836 (nuc) and rs3021088 localized to MT-ND2 mitochondrial gene (mito) has shown that the combination of the two led to BMI decrease (p=0.024). Each of the variants on its own does not correlate with higher BMI [p(nuc)=0.856, p(mito)=0.116)]. Although rs6701836 is intronic, it influences gene expression in the thyroid (p=0.000037). rs3021088 is a missense variant that leads to alanine to threonine substitution in the MT-ND2 gene which belongs to complex I of the electron transport chain. The analysis of the influence of genetic variants on gene expression has confirmed the trend explained above – the interaction of the two genes leads to BMI decrease (p=0.0308). Each of the mRNAs on its own is associated with higher BMI (p(mito)=0.0244 and p(nuc)=0.0269). Conclusıons: Our results show that nuclear-mitochondrial epistasis can influence BMI in T1DM patients. The correlation between transcription factor expression and mitochondrial genetic variants will be subject to further analysis.

Keywords: body mass index, epistasis, mitochondria, type 1 diabetes

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Authors: Jan Schmitt, Sophie Fischer

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After the Corona pandemic, climate change is a further, long-lasting challenge the society must deal with. An ongoing climate change need to be prevented. Nevertheless, the adoption tothe already changed climate conditionshas to be focused in many sectors. Recently, the decisive role of the economic sector with high value added can be seen in the Corona crisis. Hence, manufacturing industry as such a sector, needs to be prepared for climate change and adaption. Several examples from the manufacturing industry show the importance of a strategic effort in this field: The outsourcing of a major parts of the value chain to suppliers in other countries and optimizing procurement logistics in a time-, storage- and cost-efficient manner within a network of global value creation, can lead vulnerable impacts due to climate-related disruptions. E.g. the total damage costs after the 2011 flood disaster in Thailand, including costs for delivery failures, were estimated at 45 billion US dollars worldwide. German car manufacturers were also affected by supply bottlenecks andhave close its plant in Thailand for a short time. Another OEM must reduce the production output. In this contribution, a game-based learning approach is presented, which should enable manufacturing companies to derive their own strategies for climate adaption out of a mix of different actions. Based on data from a regional study of small, medium and large manufacturing companies in Mainfranken, a strongly industrialized region of northern Bavaria (Germany) the game-based learning approach is designed. Out of this, the actual state of efforts due to climate adaption is evaluated. First, the results are used to collect single actions for manufacturing companies and second, further actions can be identified. Then, a variety of climate adaption activities can be clustered according to the scope of activity of the company. The combination of different actions e.g. the renewal of the building envelope with regard to thermal insulation, its benefits and drawbacks leads to a specific strategy for climate adaption for each company. Within the game-based approach, the players take on different roles in a fictionalcompany and discuss the order and the characteristics of each action taken into their climate adaption strategy. Different indicators such as economic, ecologic and stakeholder satisfaction compare the success of the respective measures in a competitive format with other virtual companies deriving their own strategy. A "play through" climate change scenarios with targeted adaptation actions illustrate the impact of different actions and their combination onthefictional company.

Keywords: business strategy, climate change, climate adaption, game-based learning

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Authors: Gizaw Mamo Gebeyehu, Marianna Pap, Geza Makkai, Tibor Z. Janosi, Shima Rashidian, Tibor A. Rauch

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Sepsis poses a significant global health threat, necessitating extensive exploration of indicators tied to its pathological mechanisms and multi-organ dysfunction. While murine studies have shed light on sepsis, the intricate cellular and molecular landscape in human sepsis remains enigmatic. Exploring the influence of activated monocyte-derived exosomes in sepsis sheds light on a promising pathway for understanding the intricate cellular and molecular mechanisms involved in this condition in humans. In sepsis, exosome-borne mRNA and miRNA orchestrate immune response gene expression in recipient cells. Yet, the specifics of exosome-mediated cell-to-cell communication, especially how mRNA cargoes modulate gene expression in recipient cells, remain poorly understood. This study aims to elucidate the precise molecular pathways through which exosomal mRNA cargo, particularly MAFB, contributes to the developing sepsis-induced molecular aberrations in liver tissues, employing rigorously defined cell culture conditions. THP-1 cells were treated with LPS to induce changes in exosomal RNA profiles. Exosomes were isolated and characterized using microscopy and mass spectrometry. RNA was extracted from exosomes and sequenced. The most abundant exosomal mRNAs were subjected to GO analysis for functional annotation analysis and KEGG database analysis to identify the involved enriched pathways. PCR (Polymerase Chain Reaction), RNA sequencing, and Western blotting were involved to analyze changes in gene expression, protein levels, and signaling pathways within the liver cells( HepG2) after exposure to exosomal MAFB. This study pinpoints exosomal MAFB as a potential key regulator linked to liver cell damage during sepsis, along with associated genes (miR155HG, H3F3A, and possibly JARD2) forming a crucial molecular pathway contributing to liver cell injury, Together, these elements indicate a vital molecular pathway that plays a significant role in the emergence of liver cell injury during sepsis.. These findings suggest the importance of further research on these components for potential therapeutic interventions in managing acute liver damage in sepsis.

Keywords: sepsis, exososome, exosomal MAFB, LPS-induced THP-1 cells, RNA profiles, sepsis-triggered liver injury

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Authors: Chao Li, Lei Zhou, Ruiqi Ren, Dan Li, Yali Wang, Daxin Ni, Zijian Feng, Qun Li

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Background: In 2016, yellow fever had been first ever discovered in China, soon after the yellow fever epidemic occurred in Angola. After the discovery, China had promptly made the national protocol of control and prevention and strengthened the surveillance on passenger and vector. In this study, a descriptive analysis was conducted to summarize China’s experiences of response towards this import epidemic, in the hope of providing experiences on prevention and control of yellow fever and other similar imported infectious diseases in the future. Methods: The imported cases were discovered and reported by General Administration of Quality Supervision, Inspection and Quarantine (AQSIQ) and several hospitals. Each clinically diagnosed yellow fever case was confirmed by real-time reverse transcriptase polymerase chain reaction (RT–PCR). The data of the imported yellow fever cases were collected by local Centers for Disease Control and Prevention (CDC) through field investigations soon after they received the reports. Results: A total of 11 imported cases from Angola were reported in China, during Angola’s yellow fever outbreak. Six cases were discovered by the AQSIQ, among which two with mild symptom were initiative declarations at the time of entry. Except for one death, the remaining 10 cases all had recovered after timely and proper treatment. All cases are Chinese, and lived in Luanda, the capital of Angola. 73% were retailers (8/11) from Fuqing city in Fujian province, and the other three were labors send by companies. 10 cases had experiences of medical treatment in Luanda after onset, among which 8 cases visited the same local Chinese medicine hospital (China Railway four Bureau Hospital). Among the 11 cases, only one case had an effective vaccination. The result of emergency surveillance for mosquito density showed that only 14 containers of water were found positive around places of three cases, and the Breteau Index is 15. Conclusions: Effective response was taken to control and prevent the outbreak of yellow fever in China after discovering the imported cases. However, though the similar origin of Chinese in Angola has provided an easy access for disease detection, information sharing, health education and vaccination on yellow fever; these conveniences were overlooked during previous disease prevention methods. Besides, only one case having effective vaccination revealed the inadequate capacity of immunization service in China. These findings will provide suggestions to improve China’s capacity to deal with not only yellow fever but also other similar imported diseases in China.

Keywords: yellow fever, first import, China, suggestion

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Authors: Emma Avoyan

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The safety standards for flood protection in the Netherlands have been revised recently. It is expected that all major flood-protection structures will have to be reinforced to meet the new standards. The Dutch Flood Protection Programme aims at accomplishing this task through innovative integrated projects such as construction of multi-functional flood defenses. In these projects, flood safety purposes will be combined with spatial planning, nature development, emergency management or other sectoral objectives. Therefore, implementation of dike reinforcement projects requires early involvement and collaboration between public and private sectors, different governmental actors and agencies. The development and implementation of such integrated projects has been an issue in Dutch flood risk management since long. Therefore, this article analyses how cross-sector collaboration within flood risk governance in the Netherlands has evolved over time, and how this development can be explained. The integrative framework for collaborative governance is applied as an analytical tool to map external factors framing possibilities as well as constraints for cross-sector collaboration in Dutch flood risk domain. Supported by an extensive document and literature analysis, the paper offers insights on how the system context and different drivers changing over time either promoted or hindered cross-sector collaboration between flood protection sector, urban development, nature conservation or any other sector involved in flood risk governance. The system context refers to the multi-layered and interrelated suite of conditions that influence the formation and performance of complex governance systems, such as collaborative governance regimes, whereas the drivers initiate and enable the overall process of collaboration. In addition, by applying a method of process tracing we identify a causal and chronological chain of events shaping cross-sectoral interaction in Dutch flood risk management. Our results indicate that in order to evaluate the performance of complex governance systems, it is important to firstly study the system context that shapes it. Clear understanding of the system conditions and drivers for collaboration gives insight into the possibilities of and constraints for effective performance of complex governance systems. The performance of the governance system is affected by the system conditions, while at the same time the governance system can also change the system conditions. Our results show that the sequence of changes within the system conditions and drivers over time affect how cross-sector interaction in Dutch flood risk governance system happens now. Moreover, we have traced the potential of this governance system to shape and change the system context.

Keywords: collaborative governance, cross-sector interaction, flood risk management, the Netherlands

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Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

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MicroRNAs (miRNA) are a class of endogenous, single-stranded, small, and non-protein coding RNA molecules typically 20-25 nucleotides long. They are thought to regulate the expression of other genes in a broad range by binding to 3’- untranslated regions (3’-UTRs) of specific mRNAs. The detection of miRNAs is very important for understanding of the function of these molecules and in the diagnosis of variety of human diseases. However, detection of miRNAs is very challenging because of their short length and high sequence similarities within miRNA families. So, a simple-to-use, low-cost, and highly sensitive method for the detection of miRNAs is desirable. In this study, we demonstrate a novel bi-directional extension (BDE) assay. In the first step, a specific linear RT primer is hybridized to 6-10 base pairs from the 3’-end of a target miRNA molecule and then reverse transcribed to generate a cDNA strand. After reverse transcription, the cDNA was hybridized to the 3’-end which is BDE sequence; it played role as the PCR template. The PCR template was amplified in an SYBR green-based quantitative real-time PCR. To prove the concept, we used human brain total RNA. It could be detected quantitatively in the range of seven orders of magnitude with excellent linearity and reproducibility. To evaluate the performance of BDE assay, we contrasted sensitivity and specificity of the BDE assay against a commercially available poly (A) tailing method using miRNAs for let-7e extracted from A549 human epithelial lung cancer cells. The BDE assay displayed good performance compared with a poly (A) tailing method in terms of specificity and sensitivity; the CT values differed by 2.5 and the melting curve showed a sharper than poly (A) tailing methods. We have demonstrated an innovative, cost-effective BDE assay that allows improved sensitivity and specificity in detection of miRNAs. Dynamic range of the SYBR green-based RT-qPCR for miR-145 could be represented quantitatively over a range of 7 orders of magnitude from 0.1 pg to 1.0 μg of human brain total RNA. Finally, the BDE assay for detection of miRNA species such as let-7e shows good performance compared with a poly (A) tailing method in terms of specificity and sensitivity. Thus BDE proves a simple, low cost, and highly sensitive assay for various miRNAs and should provide significant contributions in research on miRNA biology and application of disease diagnostics with miRNAs as targets.

Keywords: bi-directional extension (BDE), microRNA (miRNA), poly (A) tailing assay, reverse transcription, RT-qPCR

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Authors: Santi Maneewatchararangsri, Wanpen Chaicumpa, Patcharin Saengjaruk, Urai Chaisri

Abstract:

Leptospirosis is a globally neglected disease that continues to be a significant public health and veterinary burden, with millions of cases reported each year. Early and accurate differential diagnosis of leptospirosis from other febrile illnesses and the development of a broad spectrum of leptospirosis vaccines are needed. The LipL32 outer membrane lipoprotein is a member of Leptospira adhesive matrices and has been found to exert hemolytic activity to erythrocytes in vitro. Therefore, LipL32 is regarded as a potential target for diagnosis, broad-spectrum leptospirosis vaccines, and for passive immunotherapy. In this study, we established LipL32-specific mouse monoclonal antibodies, mAbLPF1 and mAbLPF2, and their respective mouse- and humanized-engineered single chain variable fragment (ScFv). Their antibodies’ neutralizing activities against Leptospira-mediated hemolysis in vitro, and the therapeutic efficacy of mAbs against heterologous Leptospira infected hamsters were demonstrated. The epitope peptide of mAb LPF1 was mapped to a non-contiguous carboxy-terminal β-turn and amphipathic α-helix of LipL32 structure contributing to phospholipid/host cell adhesion and membrane insertion. We found that the mAbLPF2 epitope was located on the interacting loop of peptide binding groove of the LipL32 molecule responsible for interactions with host constituents. Epitope sequences are highly conserved among Leptospira spp. and are absent from the LipL32 superfamily of other microorganisms. Both epitopes are surface-exposed, readily accessible by mAbs, and immunogenic. However, they are less dominant when revealed by LipL32-specific immunoglobulins from leptospirosis-patient sera and rabbit hyperimmune serum raised by whole Leptospira. Our study also demonstrated an adhesion inhibitory activity of LipL32 protein to host membrane components and cells mediated by mAbs as well as an anti-hemolytic activity of the respective antibodies. The therapeutic antibodies, particularly the humanized-ScFv, have a potential for further development as non-drug therapeutic agent for human leptospirosis, especially in subjects allergic to antibiotics. The epitope peptides recognized by two therapeutic mAbs have potential use as tools for structure-function studies. Finally, protective peptides may be used as a target for epitope-based vaccines for control of leptospirosis.

Keywords: leptospira lipl32-specific antibodies, therapeutic epitopes, epitopes characterization, immunotherapy

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