Search results for: Heyd–Scuseria–Ernzerhof (HSE) hybrid functional

Authors: Thi Thi Zin, Pyke Tin, Ikuo Kobayashi, Yoichiro Horii

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Today dairy farm experts and farmers have well recognized the importance of dairy cow Body Condition Score (BCS) since these scores can be used to optimize milk production, managing feeding system and as an indicator for abnormality in health even can be utilized to manage for having healthy calving times and process. In tradition, BCS measures are done by animal experts or trained technicians based on visual observations focusing on pin bones, pin, thurl and hook area, tail heads shapes, hook angles and short and long ribs. Since the traditional technique is very manual and subjective, the results can lead to different scores as well as not cost effective. Thus this paper proposes an algebraic geometric imaging approach for an automatic dairy cow BCS system. The proposed system consists of three functional modules. In the first module, significant landmarks or anatomical points from the cow image region are automatically extracted by using image processing techniques. To be specific, there are 23 anatomical points in the regions of ribs, hook bones, pin bone, thurl and tail head. These points are extracted by using block region based vertical and horizontal histogram methods. According to animal experts, the body condition scores depend mainly on the shape structure these regions. Therefore the second module will investigate some algebraic and geometric properties of the extracted anatomical points. Specifically, the second order polynomial regression is employed to a subset of anatomical points to produce the regression coefficients which are to be utilized as a part of feature vector in scoring process. In addition, the angles at thurl, pin, tail head and hook bone area are computed to extend the feature vector. Finally, in the third module, the extracted feature vectors are trained by using Markov Classification process to assign BCS for individual cows. Then the assigned BCS are revised by using multiple regression method to produce the final BCS score for dairy cows. In order to confirm the validity of proposed method, a monitoring video camera is set up at the milk rotary parlor to take top view images of cows. The proposed method extracts the key anatomical points and the corresponding feature vectors for each individual cows. Then the multiple regression calculator and Markov Chain Classification process are utilized to produce the estimated body condition score for each cow. The experimental results tested on 100 dairy cows from self-collected dataset and public bench mark dataset show very promising with accuracy of 98%.

Keywords: algebraic geometric imaging approach, body condition score, Markov classification, polynomial regression

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Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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Authors: Natalia V. Beloglazova, Pieterjan Lenain, Martin Hedstrom, Dietmar Knopp, Sarah De Saeger

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In recent years polycyclic aromatic hydrocarbons (PAHs), which represent a hazard to humans and entire ecosystem, have been receiving an increased interest due to their mutagenic, carcinogenic and endocrine disrupting properties. They are formed in all incomplete combustion processes of organic matter and, as a consequence, ubiquitous in the environment. Benzo(a)pyrene (BaP) is on the priority list published by the Environmental Agency (US EPA) as the first PAH to be identified as a carcinogen and has often been used as a marker for PAHs contamination in general. It can be found in different types of water samples, therefore, the European Commission set up a limit value of 10 ng L–1 (10 ppt) for BAP in water intended for human consumption. Generally, different chromatographic techniques are used for PAHs determination, but these assays require pre-concentration of analyte, create large amounts of solvent waste, and are relatively time consuming and difficult to perform on-site. An alternative robust, stand-alone, and preferably cheap solution is needed. For example, a sensing unit which can be submerged in a river to monitor and continuously sample BaP. An affinity sensor based on capacitive transduction was developed. Natural antibodies or their synthetic analogues can be used as ligands. Ideally the sensor should operate independently over a longer period of time, e.g. several weeks or months, therefore the use of molecularly imprinted polymers (MIPs) was discussed. MIPs are synthetic antibodies which are selective for a chosen target molecule. Their robustness allows application in environments for which biological recognition elements are unsuitable or denature. They can be reused multiple times, which is essential to meet the stand-alone requirement. BaP is a highly lipophilic compound and does not contain any functional groups in its structure, thus excluding non-covalent imprinting methods based on ionic interactions. Instead, the MIPs syntheses were based on non-covalent hydrophobic and π-π interactions. Different polymerization strategies were compared and the best results were demonstrated by the MIPs produced using electropolymerization. 4-vinylpyridin (VP) and divinylbenzene (DVB) were used as monomer and cross-linker in the polymerization reaction. The selectivity and recovery of the MIP were compared to a non-imprinted polymer (NIP). Electrodes were functionalized with natural receptor (monoclonal anti-BaP antibody) and with MIPs selective towards BaP. Different sets of electrodes were evaluated and their properties such as sensitivity, selectivity and linear range were determined and compared. It was found that both receptor can reach the cut-off level comparable to the established ML, and despite the fact that the antibody showed the better cross-reactivity and affinity, MIPs were more convenient receptor due to their ability to regenerate and stability in river till 7 days.

Keywords: antibody, benzo(a)pyrene, capacitive sensor, MIPs, river water

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Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

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Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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Authors: Oliver Bertram, Leonel Akoto Chama

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In today's civil transport aircraft, the design of flight control systems is based on the experience gained from previous aircraft configurations with a clear distinction between primary and secondary flight control functions for controlling the aircraft altitude and trajectory. Significant system improvements are now seen particularly in multifunctional moveable concepts where the flight control functions are no longer considered separate but integral. This allows new functions to be implemented in order to improve the overall aircraft performance. However, the classical design process of flight controls is sequential and insufficiently interdisciplinary. In particular, the systems discipline is involved only rudimentarily in the early phase. In many cases, the task of systems design is limited to meeting the requirements of the upstream disciplines, which may lead to integration problems later. For this reason, approaching design with an incremental development is required to reduce the risk of a complete redesign. Although the potential and the path to multifunctional moveable concepts are shown, the complete re-engineering of aircraft concepts with less classic moveable concepts is associated with a considerable risk for the design due to the lack of design methods. This represents an obstacle to major leaps in technology. This gap in state of the art is even further increased if, in the future, unconventional aircraft configurations shall be considered, where no reference data or architectures are available. This means that the use of the above-mentioned experience-based approach used for conventional configurations is limited and not applicable to the next generation of aircraft. In particular, there is a need for methods and tools for a rapid trade-off between new multifunctional flight control systems architectures. To close this gap in the state of the art, an integrated system-driven design process for multifunctional flight control systems of non-classical aircraft configurations will be presented. The overall goal of the design process is to find optimal solutions for single or combined target criteria in a fast process from the very large solution space for the flight control system. In contrast to the state of the art, all disciplines are involved for a holistic design in an integrated rather than a sequential process. To emphasize the systems discipline, this paper focuses on the methodology for designing moveable actuation systems in the context of this integrated design process of multifunctional moveables. The methodology includes different approaches for creating system architectures, component design methods as well as the necessary process outputs to evaluate the systems. An application example of a reference configuration is used to demonstrate the process and validate the results. For this, new unconventional hydraulic and electrical flight control system architectures are calculated which result from the higher requirements for multifunctional moveable concept. In addition to typical key performance indicators such as mass and required power requirements, the results regarding the feasibility and wing integration aspects of the system components are examined and discussed here. This is intended to show how the systems design can influence and drive the wing and overall aircraft design.

Keywords: actuation systems, flight control surfaces, multi-functional movables, wing design process

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Authors: Igor Guatelli

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The universe of aesthetic perception entails impasses about sensitive divergences that each text or visual object may be subjected to. If approached through intertextuality that is not based on the misleading notion of kinships or similarities a priori admissible, the possibility of anachronistic, heterogeneous - and non-diachronic - assemblies can enhance the emergence of interval movements, intermediate, and conflicting, conducive to a method of reading, interpreting, and assigning meaning that escapes the rigid antinomies of the mere being and non-being of things. In negative, they operate in a relationship built by the lack of an adjusted meaning set by their positive existences, with no remainders; the generated interval becomes the remnant of each of them; it is the opening that obscures the stable positions of each one. Without the negative of absence, of that which is always missing or must be missing in a text, concept, or image made positive by history, nothing is perceived beyond what has been already given. Pairings or binary oppositions cannot lead only to functional syntheses; on the contrary, methodological disturbances accumulated by the approximation of signs and entities can initiate a process of becoming as an opening to an unforeseen other, transformation until a moment when the difficulties of [re]conciliation become the mainstay of a future of that sign/entity, not envisioned a priori. A counter-history can emerge from these unprecedented, misadjusted approaches, beginnings of unassigned injunctions and disjunctions, in short, difficult alliances that open cracks in a supposedly cohesive history, chained in its apparent linearity with no remains, understood as a categorical historical imperative. Interstices are minority fields that, because of their opening, are capable of causing opacity in that which, apparently, presents itself with irreducible clarity. Resulting from an incomplete and maladjusted [at the least dual] marriage between the signs/entities that originate them, this interval may destabilize and cause disorder in these entities and their own meanings. The interstitials offer a hyphenated relationship: a simultaneous union and separation, a spacing between the entity’s identity and its otherness or, alterity. One and the other may no longer be seen without the crack or fissure that now separates them, uniting, by a space-time lapse. Ontological, semantic shifts are caused by this fissure, an absence between one and the other, one with and against the other. Based on an improbable approximation between some conceptual and semantic shifts within the design production of architect Rem Koolhaas and the textual production of the philosopher Jacques Derrida, this article questions the notion of unity, coherence, affinity, and complementarity in the process of construction of thought from these ontological, epistemological, and semiological fissures that rattle the signs/entities and their stable meanings. Fissures in a thought that is considered coherent, cohesive, formatted are the negativity that constitutes the interstices that allow us to move towards what still remains as non-identity, which allows us to begin another story.

Keywords: clearing, interstice, negative, remnant, spectrum

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Authors: Rozh Abdulrahman Kareem

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The displacement of individuals has become a common interest for international players. Mostly occurs in Islamic states, as religion is considered the most common cause of this form of displacement. Therefore, this thesis aims to depict the reality of the situations of the refugees, particularly in KRI, illustrating how they are treated and protected and if the treatment merits the protection clause as envisaged in the 1951 Refugee Convention. Overall, the aim is to touch on the issue of protection by non-governmental organizations and government towards the refugees here. In light of this, it focused on the adequate protection of refugees in relation to the refugee law. In the Middle East, including Iraq, there have been multiple reports on violations of these refugee laws and human rights. Protection involves providing physical security to the concerned parties, functional administration with legal structures, and infrastructural setup that could help citizens exercise rights. The KRI has provided the refugees with various benefits, including education, access to residency, and employment. It also provided transitionary in various social dimensions like gender-based violence. The Convention on Status of Refugees 1951 tried to resolve this problem, whereby the principle of ‘nonrefoulement’ under Article 33 was passed. The ‘nonrefoulement’, an exceptional reference, was enacted to protect refugees from forcible return to their countries of the original. However, the convention never addressed an unusual scenario regarding the application of this principle, ‘Extradition Treaties.’ Even though some scholarly article exists regarding the problems of refugees, the topic of interplay between Nonrefoulement and Extradition Treaties has never been explained in detail in the available books on refugee laws and practices. Each year, millions of refugees seek protection from foreign countries for fear of being tortured, victimized, or executed. People seeking international protection are susceptible and insecure. The main objective of the prevention is to provide security to citizens susceptible to inhuman treatment, distress, oppression, or other human rights defilements when they arrive back in their own countries. The refugee situation may get worse in the near future. Just like several nations within the Middle East, Iraq is not a signatory to the globally acknowledged legal structure for the protection of refugees. The first law of 1971 in Iraq was issued only for military or political causes. This law also establishes benefits such as the right to education and health services and the right to acquire employment just as the Iraqi nationals. The other legislative instrument is the 21st law from the ministry of migration of Iraq widened the description of an immigrant to incorporate the definition from the refugee resolution. Nonetheless, there is a lack of overall consistency in the protection provided under these legislations regarding rights and entitlement. A Memorandum of Understanding was signed in October 2016 by the UNHCR and the Iraq government to develop the protection of refugees. Under the term of this MoU, the Iraqi Government is obligated to provide identity documents to asylum seekers beside that UNHCR provides more guidance.

Keywords: law, refugee, protection, Kurdistan

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Authors: Thauane Silva, Gabrielle do Vale, Andre Ferreira, Marilda Siqueira, Thiago Moreno L. Souza, Milene D. Miranda

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The exposure of A(H1N1)pdm09-infected epithelial cells (HeLa) to HIV-1 viral particles, or its gp120, enhanced interferon-induced transmembrane protein (IFITM3) content, a viral restriction factor (RF), resulting in a decrease in influenza replication. The gp120 binds to CCR5 (R5) or CXCR4 (X4) cell receptors during HIV-1 infection. Then, it is possible that the endogenous ligands of these receptors also modulate the expression of IFITM3 and other cellular factors that restrict influenza virus replication. Thus, the aim of this study is to analyze the role of cellular receptors R5 and X4 in modulating RFs in order to inhibit the replication of the influenza virus. A549 cells were treated with 2x effective dose (ED50) of endogenous R5 or X4 receptor agonists, CCL3 (20 ng/ml), CCL4 (10 ng/ml), CCL5 (10 ng/ml) and CXCL12 (100 ng/mL) or exogenous agonists, gp120 Bal-R5, gp120 IIIB-X4 and its mutants (5 µg/mL). The interferon α (10 ng/mL) and oseltamivir (60 nM) were used as a control. After 24 h post agonists exposure, the cells were infected with virus influenza A(H3N2) at 2 MOI (multiplicity of infection) for 1 h. Then, 24 h post infection, the supernatant was harvested and, the viral titre was evaluated by qRT-PCR. To evaluate IFITM3 and SAM and HD domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (SAMHD1) protein levels, A549 were exposed to agonists for 24 h, and the monolayer was lysed with Laemmli buffer for western blot (WB) assay or fixed for indirect immunofluorescence (IFI) assay. In addition to this, we analyzed other RFs modulation in A549, after 24 h post agonists exposure by customized RT² Profiler Polymerase Chain Reaction Array. We also performed a functional assay in which SAMHD1-knocked-down, by single-stranded RNA (siRNA), A549 cells were infected with A(H3N2). In addition, the cells were treated with guanosine to assess the regulatory role of dNTPs by SAMHD1. We found that R5 and X4 agonists inhibited influenza replication in 54 ± 9%. We observed a four-fold increase in SAMHD1 transcripts by RFs mRNA quantification panel. After 24 h post agonists exposure, we did not observe an increase in IFITM3 protein levels through WB or IFI assays, but we observed an upregulation up to three-fold in the protein content of SAMHD1, in A549 exposed to agonists. Besides this, influenza replication enhanced in 20% in cell cultures that SAMDH1 was knockdown. Guanosine treatment in cells exposed to R5 ligands further inhibited influenza virus replication, suggesting that the inhibitory mechanism may involve the activation of the SAMHD1 deoxynucleotide triphosphohydrolase activity. Thus, our data show for the first time a direct relationship of SAMHD1 and inhibition of influenza replication, and provides perspectives for new studies on the signaling modulation, through cellular receptors, to induce proteins of great importance in the control of relevant infections for public health.

Keywords: chemokine receptors, gp120, influenza, virus restriction factors

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Authors: Muhammad Naveed Tahir, Wang Yingkuan, Huang Wenjiang, Raheel Osman

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Numerous models used in prediction and decision-making process but most of them are linear in natural environment, and linear models reach their limitations with non-linearity in data. Therefore accurate estimation is difficult. Artificial Neural Networks (ANN) found extensive acceptance to address the modeling of the complex real world for the non-linear environment. ANN’s have more general and flexible functional forms than traditional statistical methods can effectively deal with. The link between information technology and agriculture will become more firm in the near future. Monitoring crop biophysical properties non-destructively can provide a rapid and accurate understanding of its response to various environmental influences. Crop chlorophyll content is an important indicator of crop health and therefore the estimation of crop yield. In recent years, remote sensing has been accepted as a robust tool for site-specific management by detecting crop parameters at both local and large scales. The present research combined the ANN model with satellite-derived chlorophyll indices from LANDSAT 8 imagery for predicting real-time wheat chlorophyll estimation. The cloud-free scenes of LANDSAT 8 were acquired (Feb-March 2016-17) at the same time when ground-truthing campaign was performed for chlorophyll estimation by using SPAD-502. Different vegetation indices were derived from LANDSAT 8 imagery using ERADAS Imagine (v.2014) software for chlorophyll determination. The vegetation indices were including Normalized Difference Vegetation Index (NDVI), Green Normalized Difference Vegetation Index (GNDVI), Chlorophyll Absorbed Ratio Index (CARI), Modified Chlorophyll Absorbed Ratio Index (MCARI) and Transformed Chlorophyll Absorbed Ratio index (TCARI). For ANN modeling, MATLAB and SPSS (ANN) tools were used. Multilayer Perceptron (MLP) in MATLAB provided very satisfactory results. For training purpose of MLP 61.7% of the data, for validation purpose 28.3% of data and rest 10% of data were used to evaluate and validate the ANN model results. For error evaluation, sum of squares error and relative error were used. ANN model summery showed that sum of squares error of 10.786, the average overall relative error was .099. The MCARI and NDVI were revealed to be more sensitive indices for assessing wheat chlorophyll content with the highest coefficient of determination R²=0.93 and 0.90 respectively. The results suggested that use of high spatial resolution satellite imagery for the retrieval of crop chlorophyll content by using ANN model provides accurate, reliable assessment of crop health status at a larger scale which can help in managing crop nutrition requirement in real time.

Keywords: ANN, chlorophyll content, chlorophyll indices, satellite images, wheat

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Authors: Christoph Opperer

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The rapid urbanization of modern societies and the need for sustainable urban development demand innovative solutions that meet both individual and collective needs while addressing environmental concerns. To address these challenges, this paper presents a study that explores the potential of spatial and energetic/ecological optimization to enhance the performance of urban settlements, focusing on both architectural and urban scales. The study focuses on the application of biological principles and self-organization processes in urban planning and design, aiming to achieve a balance between ecological performance, architectural quality, and individual living conditions. The research adopts a case study approach, focusing on a 10-hectare brownfield site in the south of Vienna. The site is surrounded by a small-scale built environment as an appropriate starting point for the research and design process. However, the selected urban form is not a prerequisite for the proposed design methodology, as the findings can be applied to various urban forms and densities. The methodology used in this research involves dividing the overall building mass and program into individual small housing units. A computational model has been developed to optimize the distribution of these units, considering factors such as solar exposure/radiation, views, privacy, proximity to sources of disturbance (such as noise), and minimal internal circulation areas. The model also ensures that existing vegetation and buildings on the site are preserved and incorporated into the optimization and design process. The model allows for simultaneous optimization at two scales, architectural and urban design, which have traditionally been addressed sequentially. This holistic design approach leads to individual and collective benefits, resulting in urban environments that foster a balance between ecology and architectural quality. The results of the optimization process demonstrate a seemingly random distribution of housing units that, in fact, is a densified hybrid between traditional garden settlements and allotment settlements. This urban typology is selected due to its compatibility with the surrounding urban context, although the presented methodology can be extended to other forms of urban development and density levels. The benefits of this approach are threefold. First, it allows for the determination of ideal housing distribution that optimizes solar radiation for each building density level, essentially extending the concept of sustainable building to the urban scale. Second, the method enhances living quality by considering the orientation and positioning of individual functions within each housing unit, achieving optimal views and privacy. Third, the algorithm's flexibility and robustness facilitate the efficient implementation of urban development with various stakeholders, architects, and construction companies without compromising its performance. The core of the research is the application of global and local optimization strategies to create efficient design solutions. By considering both, the performance of individual units and the collective performance of the urban aggregation, we ensure an optimal balance between private and communal benefits. By promoting a holistic understanding of urban ecology and integrating advanced optimization strategies, our methodology offers a sustainable and efficient solution to the challenges of modern urbanization.

Keywords: sustainable development, self-organization, ecological performance, solar radiation and exposure, daylight, visibility, accessibility, spatial distribution, local and global optimization

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Authors: Linyi Fan, C.J. Schumaker

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Background: Biomedical efforts to treat Alzheimer’s disease (AD) have typically produced mixed to poor results, while more lifestyle-focused treatments such as exercise may fare better than existing biomedical treatments. A few promising studies have indicated that activities of daily life (ADL) may be a useful way of predicting AD. However, the existing cross-sectional studies fail to show how functional-related issues such as ADL in early years predict AD and how social factors influence health either in addition to or in interaction with individual risk factors. This study would helpbetterscreening and early treatments for the elderly population and healthcare practice. The findings have significance academically and practically in terms of creating positive social change. Methodology: The purpose of this quantitative historical, correlational study was to examine the relationship between early years’ ADL and the development of AD in later years. The studyincluded 4,526participantsderived fromRAND HRS dataset. The Health and Retirement Study (HRS) is a longitudinal household survey data set that is available forresearchof retirement and health among the elderly in the United States. The sample was selected by the completion of survey questionnaire about AD and dementia. The variablethat indicates whether the participant has been diagnosed with AD was the dependent variable. The ADL indices and changes in ADL were the independent variables. A four-step multilevel regression model approach was utilized to address the research questions. Results: Amongst 4,526 patients who completed the AD and dementia questionnaire, 144 (3.1%) were diagnosed with AD. Of the 4,526 participants, 3,465 (76.6%) have high school and upper education degrees,4,074 (90.0%) were above poverty threshold. The model evaluatedthe effect of ADL and change in ADL on onset of AD in late years while allowing the intercept of the model to vary by level of education. The results suggested that the only significant predictor of the onset of AD was changes in early years’ ADL (b = 20.253, z = 2.761, p < .05). However, the result of the sensitivity analysis (b = 7.562, z = 1.900, p =.058), which included more control variables and increased the observation period of ADL, are not supported this finding. The model also estimated whether the variances of random effect vary by Level-2 variables. The results suggested that the variances associated with random slopes were approximately zero, suggesting that the relationship between early years’ ADL were not influenced bysociodemographic factors. Conclusion: The finding indicated that an increase in changes in ADL leads to an increase in the probability of onset AD in the future. However, this finding is not support in a broad observation period model. The study also failed to reject the hypothesis that the sociodemographic factors explained significant amounts of variance in random effect. Recommendations were then made for future research and practice based on these limitations and the significance of the findings.

Keywords: alzheimer’s disease, epidemiology, moderation, multilevel modeling

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Authors: Elham Koohi, Silvio Jose Gumiere, Hossein Bonakdari, Saeid Homayouni

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Water used in agricultural crops can be managed by irrigation scheduling based on soil moisture levels and plant water stress thresholds. Automated irrigation scheduling limits crop physiological damage and yield reduction. Knowledge of crop water stress monitoring approaches can be effective in optimizing the use of agricultural water. Understanding the physiological mechanisms of crop responding and adapting to water deficit ensures sustainable agricultural management and food supply. This aim could be achieved by analyzing and diagnosing crop characteristics and their interlinkage with the surrounding environment. Assessments of plant functional types (e.g., leaf area and structure, tree height, rate of evapotranspiration, rate of photosynthesis), controlling changes, and irrigated areas mapping. Calculating thresholds of soil water content parameters, crop water use efficiency, and Nitrogen status make irrigation scheduling decisions more accurate by preventing water limitations between irrigations. Combining Remote Sensing (RS), the Internet of Things (IoT), Artificial Intelligence (AI), and Machine Learning Algorithms (MLAs) can improve measurement accuracies and automate irrigation scheduling. This paper is a review structured by surveying about 100 recent research studies to analyze varied approaches in terms of providing high spatial and temporal resolution mapping, sensor-based Variable Rate Application (VRA) mapping, the relation between spectral and thermal reflectance and different features of crop and soil. The other objective is to assess RS indices formed by choosing specific reflectance bands and identifying the correct spectral band to optimize classification techniques and analyze Proximal Optical Sensors (POSs) to control changes. The innovation of this paper can be defined as categorizing evaluation methodologies of precision irrigation (applying the right practice, at the right place, at the right time, with the right quantity) controlled by soil moisture levels and sensitiveness of crops to water stress, into pre-processing, processing (retrieval algorithms), and post-processing parts. Then, the main idea of this research is to analyze the error reasons and/or values in employing different approaches in three proposed parts reported by recent studies. Additionally, as an overview conclusion tried to decompose different approaches to optimizing indices, calibration methods for the sensors, thresholding and prediction models prone to errors, and improvements in classification accuracy for mapping changes.

Keywords: agricultural crops, crop water stress detection, irrigation scheduling, precision agriculture, remote sensing

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Authors: Angelis P. Barlampas

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Learning Objectives: The purpose of this study is to inform briefly the reader about the applications of AI in chest radiology. Background: Currently, there are 190 FDA-approved radiology AI applications, with 42 (22%) pertaining specifically to thoracic radiology. Imaging findings OR Procedure details Aids of AI in chest radiology1: Detects and segments pulmonary nodules. Subtracts bone to provide an unobstructed view of the underlying lung parenchyma and provides further information on nodule characteristics, such as nodule location, nodule two-dimensional size or three dimensional (3D) volume, change in nodule size over time, attenuation data (i.e., mean, minimum, and/or maximum Hounsfield units [HU]), morphological assessments, or combinations of the above. Reclassifies indeterminate pulmonary nodules into low or high risk with higher accuracy than conventional risk models. Detects pleural effusion . Differentiates tension pneumothorax from nontension pneumothorax. Detects cardiomegaly, calcification, consolidation, mediastinal widening, atelectasis, fibrosis and pneumoperitoneum. Localises automatically vertebrae segments, labels ribs and detects rib fractures. Measures the distance from the tube tip to the carina and localizes both endotracheal tubes and central vascular lines. Detects consolidation and progression of parenchymal diseases such as pulmonary fibrosis or chronic obstructive pulmonary disease (COPD).Can evaluate lobar volumes. Identifies and labels pulmonary bronchi and vasculature and quantifies air-trapping. Offers emphysema evaluation. Provides functional respiratory imaging, whereby high-resolution CT images are post-processed to quantify airflow by lung region and may be used to quantify key biomarkers such as airway resistance, air-trapping, ventilation mapping, lung and lobar volume, and blood vessel and airway volume. Assesses the lung parenchyma by way of density evaluation. Provides percentages of tissues within defined attenuation (HU) ranges besides furnishing automated lung segmentation and lung volume information. Improves image quality for noisy images with built-in denoising function. Detects emphysema, a common condition seen in patients with history of smoking and hyperdense or opacified regions, thereby aiding in the diagnosis of certain pathologies, such as COVID-19 pneumonia. It aids in cardiac segmentation and calcium detection, aorta segmentation and diameter measurements, and vertebral body segmentation and density measurements. Conclusion: The future is yet to come, but AI already is a helpful tool for the daily practice in radiology. It is assumed, that the continuing progression of the computerized systems and the improvements in software algorithms , will redder AI into the second hand of the radiologist.

Keywords: artificial intelligence, chest imaging, nodule detection, automated diagnoses

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Authors: Mindaugas Liaudanskas, Darius Kviklys, Kristina Zymonė, Raimondas Raudonis, Jonas Viškelis, Norbertas Uselis, Pranas Viškelis, Valdimaras Janulis

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Apples are among the most commonly consumed fruits in the world. Based on data from the year 2014, approximately 84.63 million tons of apples are grown per annum. Apples are widely used in food industry to produce various products and drinks (juice, wine, and cider); they are also used unprocessed. Apples in human diet are an important source of different groups of biological active compounds that can positively contribute to the prevention of various diseases. They are a source of various biologically active substances – especially vitamins, organic acids, micro- and macro-elements, pectins, and phenolic, triterpenic, and other compounds. Triterpenic compounds, which are characterized by versatile biological activity, are the biologically active compounds found in apples that are among the most promising and most significant for human health. A specific analytical procedure including sample preparation and High Performance Liquid Chromatography (HPLC) analysis was developed, optimized, and validated for the detection of triterpenic compounds in the samples of different apples, their peels, and flesh from widespread apple cultivars 'Aldas', 'Auksis', 'Connel Red', 'Ligol', 'Lodel', and 'Rajka' grown in Lithuanian climatic conditions. The conditions for triterpenic compound extraction were optimized: the solvent of the extraction was 100% (v/v) acetone, and the extraction was performed in an ultrasound bath for 10 min. Isocratic elution (the eluents ratio being 88% (solvent A) and 12% (solvent B)) for a rapid separation of triterpenic compounds was performed. The validation of the methodology was performed on the basis of the ICH recommendations. The following characteristics of validation were evaluated: the selectivity of the method (specificity), precision, the detection and quantitation limits of the analytes, and linearity. The obtained parameters values confirm suitability of methodology to perform analysis of triterpenic compounds. Using the optimised and validated HPLC technique, four triterpenic compounds were separated and identified, and their specificity was confirmed. These compounds were corosolic acid, betulinic acid, oleanolic acid, and ursolic acid. Ursolic acid was the dominant compound in all the tested apple samples. The detected amount of betulinic acid was the lowest of all the identified triterpenic compounds. The greatest amounts of triterpenic compounds were detected in whole apple and apple peel samples of the 'Lodel' cultivar, and thus apples and apple extracts of this cultivar are potentially valuable for use in medical practice, for the prevention of various diseases, for adjunct therapy, for the isolation of individual compounds with a specific biological effect, and for the development and production of dietary supplements and functional food enriched in biologically active compounds. Acknowledgements. This work was supported by a grant from the Research Council of Lithuania, project No. MIP-17-8.

Keywords: apples, HPLC, triterpenic compounds, validation

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Authors: Saleh Gorji, Sahil Gulati, Ana Pakzad

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Continuous electron diffraction tomography, also known as microcrystal electron diffraction (MicroED) or three-dimensional electron diffraction (3DED), is a powerful technique, which in combination with cryo-electron microscopy (cryo-ED), can provide atomic-scale 3D information about the crystal structure and composition of different classes of crystalline materials such as proteins, peptides, and small molecules. Unlike the well-established X-ray crystallography method, 3DED does not require large single crystals and can collect accurate electron diffraction data from crystals as small as 50 – 100 nm. This is a critical advantage as growing larger crystals, as required by X-ray crystallography methods, is often very difficult, time-consuming, and expensive. In most cases, specimens studied via 3DED method are electron beam sensitive, which means there is a limitation on the maximum amount of electron dose one can use to collect the required data for a high-resolution structure determination. Therefore, collecting data using a conventional scintillator-based fiber coupled camera brings additional challenges. This is because of the inherent noise introduced during the electron-to-photon conversion in the scintillator and transfer of light via the fibers to the sensor, which results in a poor signal-to-noise ratio and requires a relatively higher and commonly specimen-damaging electron dose rates, especially for protein crystals. As in other cryo-EM techniques, damage to the specimen can be mitigated if a direct detection camera is used which provides a high signal-to-noise ratio at low electron doses. In this work, we have used two classes of such detectors from Gatan, namely the K3® camera (a monolithic active pixel sensor) and Stela™ (that utilizes DECTRIS hybrid-pixel technology), to address this problem. The K3 is an electron counting detector optimized for low-dose applications (like structural biology cryo-EM), and Stela is also a counting electron detector but optimized for diffraction applications with high speed and high dynamic range. Lastly, data collection workflows, including crystal screening, microscope optics setup (for imaging and diffraction), stage height adjustment at each crystal position, and tomogram acquisition, can be one of the other challenges of the 3DED technique. Traditionally this has been all done manually or in a partly automated fashion using open-source software and scripting, requiring long hours on the microscope (extra cost) and extensive user interaction with the system. We have recently introduced Latitude® D in DigitalMicrograph® software, which is compatible with all pre- and post-energy-filter Gatan cameras and enables 3DED data acquisition in an automated and optimized fashion. Higher quality 3DED data enables structure determination with higher confidence, while automated workflows allow these to be completed considerably faster than before. Using multiple examples, this work will demonstrate how to direct detection electron counting cameras enhance 3DED results (3 to better than 1 Angstrom) for protein and small molecule structure determination. We will also show how Latitude D software facilitates collecting such data in an integrated and fully automated user interface.

Keywords: continuous electron diffraction tomography, direct detection, diffraction, Latitude D, Digitalmicrograph, proteins, small molecules

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Authors: Lina Paola Orozco Marin, Yuliet Montoya Osorio, John Bustamante Osorno

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Ischemic events can culminate in acute myocardial infarction by irreversible cardiac lesions that cannot be restored due to the limited regenerative capacity of the heart. Cell therapy seeks to replace these injured or necrotic cells by transplanting healthy and functional cells. The therapeutic alternatives proposed by tissue engineering and cardiovascular regenerative medicine are the use of biomaterials to mimic the native extracellular medium, which is full of proteins, proteoglycans, and glycoproteins. The selected biomaterials must provide structural support to the encapsulated cells to avoid their migration and death in the host tissue. In this context, the present research work focused on developing a natural thermosensitive hydrogel, its physical and chemical characterization, and the determination of its biocompatibility in vitro. The hydrogel was developed by mixing hydrolyzed bovine and porcine collagen at 2% w/v, chitosan at 2.5% w/v, and beta-glycerolphosphate at 8.5% w/w and 10.5% w/w in magnetic stirring at 4°C. Once obtained, the thermosensitivity and gelation time were determined, incubating the samples at 37°C and evaluating them through the inverted tube method. The morphological characterization of the hydrogels was carried out through scanning electron microscopy. Chemical characterization was carried out employing infrared spectroscopy. The biocompatibility was determined using the MTT cytotoxicity test according to the ISO 10993-5 standard for the hydrogel’s precursors using the fetal human ventricular cardiomyocytes cell line RL-14. The RL-14 cells were also seeded on the top of the hydrogels, and the supernatants were subculture at different periods to their observation under a bright field microscope. Four types of thermosensitive hydrogels were obtained, which differ in their composition and concentration, called A1 (chitosan/bovine collagen/beta-glycerolphosphate 8.5%w/w), A2 (chitosan/porcine collagen/beta-glycerolphosphate 8.5%), B1 (chitosan/bovine collagen/beta-glycerolphosphate 10.5%) and B2 (chitosan/porcine collagen/beta-glycerolphosphate 10.5%). A1 and A2 had a gelation time of 40 minutes, and B1 and B2 had a gelation time of 30 minutes at 37°C. Electron micrographs revealed a three-dimensional internal structure with interconnected pores for the four types of hydrogels. This facilitates the exchange of nutrients, oxygen, and the exit of metabolites, allowing to preserve a microenvironment suitable for cell proliferation. In the infrared spectra, it was possible to observe the interaction that occurs between the amides of polymeric compounds with the phosphate groups of beta-glycerolphosphate. Finally, the biocompatibility tests indicated that cells in contact with the hydrogel or with each of its precursors are not affected in their proliferation capacity for a period of 16 days. These results show the potential of the hydrogel to increase the cell survival rate in the cardiac cell therapies under investigation. Moreover, the results lay the foundations for its characterization and biological evaluation in both in vitro and in vivo models.

Keywords: cardiac cell therapy, cardiac ischemia, natural polymers, thermosensitive hydrogel

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Authors: H. Alqunayeer, S. Zamir

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Blended learning, an ideal amalgamation of online learning and face to face traditional approach is a new approach that may result in outstanding outcomes in the realm of teaching and learning. The dexterity and effectiveness offered by e-learning experience cannot be guaranteed in a traditional classroom, whereas one-to-one interaction the essential element of learning that can only be found in a traditional classroom. In recent years, a spectacular expansion in the incorporation of technology in language teaching and learning is observed in many universities of Saudi Arabia. Some universities recognize the importance of blending face-to-face with online instruction in language pedagogy, Qassim University is one of the many universities adopting Blackboard Learning Management system (LMS). The university has adopted this new mode of teaching/learning in year 2015. Although the experience is immature; however great pedagogical transformations are anticipated in the university through this new approach. This paper examines the role of blended language learning with particular reference to the influence of Blackboard Learning Management System on the development of English language learning for EFL learners registered in Bachelors of English language program. This paper aims at exploring three main areas: (i) the present status of Blended learning in the educational process in Saudi Arabia especially in Qassim University by providing a survey report on the number of training courses on Blackboard LMS conducted for the male and female teachers at various colleges of Qassim University, (ii) a survey on teachers perception about the utility, application and the outcome of using blended Learning approach in teaching English language skills courses, (iii) the students’ views on the efficiency of Blended learning approach in learning English language skills courses. Besides, analysis of students’ limitations and challenges related to the experience of blended learning via Blackboard, the suggestion and recommendations offered by the language learners have also been thought-out. The study is empirical in nature. In order to gather data on the afore mentioned areas survey questionnaire method has been used: in order to study students’ perception, a 5 point Likert-scale questionnaire has been distributed to 200 students of English department registered in Bachelors in English program (level 5 through level 8). Teachers’ views have been surveyed with the help of interviewing 25 EFL teachers skilled in using Blackboard LMS in their lectures. In order to ensure the validity and reliability of questionnaire, the inter-rater approach and Cronbach’s Alpha analysis have been used respectively. Analysis of variance (ANOVA) has been used to analyze the students’ perception about the productivity of the Blended approach in learning English language skills. The analysis of feedback by Saudi teachers and students about the usefulness, ingenuity, and productivity of Blended Learning via Blackboard LMS highlights the need of encouraging and expanding the implementation of this new approach into the field of English language teaching in Saudi Arabia, in order to augment congenial learning aura. Furthermore, it is hoped that the propositions and practical suggestions offered by the study will be functional for other similar learning environments.

Keywords: blended learning, black board learning management system, English as foreign language (EFL) learners, EFL teachers

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Authors: Sundru Manjulata Devi, Prakash M. Halami

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The immemorial use of fermented foods from vegetables, dairy and other biological sources are of great demand in India because of their health benefits. However, the diversity of Lactobacillus plantarum group (LPG) of vegetable origin has not been revealed yet, particularly with reference to their probiotic functionalities. In the present study, the different species of probiotic Lactobacillus plantarum group (LPG) i.e., L. plantarum subsp. plantarum MTCC 5422 (from fermented cereals), L. plantarum subsp. argentoratensis FG16 (from fermented bamboo shoot) and L. paraplantarum MTCC 9483 (from fermented gundruk) (as characterized by multiplex recA PCR assay) were considered to investigate their relative expression of MUB domains of mub gene (mucin binding protein) by Real time PCR. Initially, the allelic variation in the mub gene was assessed and found to encode three different variants (Type I, II and III). All the three types had 8, 9 and 10 MUB domains respectively (as analysed by Pfam database) and were found to be responsible for adhesion of bacteria to the host intestinal epithelial cells. These domains either get inserted or deleted during speciation or evolutionary events and lead to divergence. The reverse transcriptase qPCR analysis with mubLPF1+R1 primer pair supported variation in amplicon sizes with 300, 500 and 700 bp among different LPG strains. The relative expression of these MUB domains significantly unregulated in the presence of 1% mucin in overnight grown cultures. Simultaneously, the mub gene expressed efficiently by 7 fold in the culture L. paraplantarum MTCC 9483 with 10 MUB domains. An increase in the expression levels for L. plantarum subsp. plantarum MTCC 5422 and L. plantarum subsp. argentoratensis FG16 (MCC 2974) with 9 and 8 repetitive domains was around 4 and 2 fold, respectively. The detection and expression of an integrase (int) gene in the upstream region of mub gene reveals the excision and integration of these repetitive domains. Concurrently, an in vitro adhesion assay to mucin and exclusion of pathogens (such as Listeria monocytogenes and Micrococcus leuteus) was investigated and observed that the L. paraplantarum MTCC 9483 with more adhesion domains has more ability to adhere to mucin and inhibited the growth of pathogens. The production and expression of plantaricin-like bacteriocin (plnNC8 type) in MTCC 9483 suggests the pathogen inhibition. Hence, the expression of MUB domains can act as potential biomarkers in the screening of a novel probiotic LPG strain with adherence property. The present study provides a platform for an easy, rapid, less time consuming, low-cost methodology for the detection of potential probiotic bacteria. It was known that the traditional practices followed in the preparation of fermented bamboo shoots/gundruk/cereals of Indian foods contain different kinds of neutraceuticals for functional food and novel compounds with health promoting factors. In future, a detailed study of these food products can add more nutritive value, consumption and suitable for commercialization.

Keywords: adhesion gene, fermented foods, MUB domains, probiotics

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Authors: Ummeh Saika, Toshio Kikuchi

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Population growth and urbanization is a global phenomenon. As the rapid progress of technology, many cities in the international community are facing serious problems of urbanization. There is no doubt that the urbanization will proceed to have significant impact on the ecology, economy and society at local, regional, and global levels. The inhabitants of Dhaka city suffer from lack of proper urban facilities. The green spaces are needed for different functional and leisure activities of the urban dwellers. Again growing densification, a number of green space are transferred into open space in the Dhaka city. As a result greenery of the city's decreases gradually. Moreover, the existing green space is frequently threatened by encroachment. The role of green space, both at community and city level, is important to improve the natural environment and social ties for future generations. Therefore, it seems that the green space needs to be more effective for public interaction. The main objective of this study is to address the effectiveness of urban green space (Urban Park) of Dhaka City. Two approaches are selected to fulfill the study. Firstly, analyze the long-term spatial changes of urban green space using GIS and secondly, investigate the relationship of urban park network with physical and social environment. The case study site covers eight urban parks of Dhaka metropolitan area of Bangladesh. Two aspects (Physical and Social) are applied for this study. For physical aspect, satellite images and aerial photos of different years are used to find out the changes of urban parks. And for social aspect, methods are used as questionnaire survey, interview, observation, photographs, sketch and previous information of parks to analyze about the social environment of parks. After calculation of all data by descriptive statistics, result is shown by maps using GIS. According to physical size, parks of Dhaka city are classified into four types: Small, Medium, Large and Extra Large parks. The observed result showed that the physical and social environment of urban parks varies with their size. In small size parks physical environment is moderate by newly tree plantation and area expansion. However, in medium size parks physical environment are poor, example- tree decrease, exposed soil increase. On the other hand, physical environment of large size and extra large size parks are in good condition, because of plenty of vegetation and well management. Again based on social environment, in small size parks people mainly come from surroundings area and mainly used as waiting place. In medium-size parks, people come to attend various occasion from different places. In large size and extra large size parks, people come from every part of the city area for tourism purpose. Urban parks are important source of green space. Its influence both physical and social environment of urban area. Nowadays green space area gradually decreases and transfer into open space. The consequence of this research reveals that changes of urban parks influence both physical and social environment and also impact on urban life.

Keywords: physical environment, social environment, urban life, urban parks

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Authors: L. P. Gomes, G. F. Araújo, Y. M. L. Cordeiro, C. T. Andrade, E. M. Del Aguila, V. M. F. Paschoalin

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The application of nanoscale materials and nanostructures is an emerging area, these since materials may provide solutions to technological and environmental challenges in order to preserve the environment and natural resources. To reach this goal, the increasing demand must be accompanied by 'green' synthesis methods. Chitosan is a natural, nontoxic, biopolymer derived by the deacetylation of chitin and has great potential for a wide range of applications in the biological and biomedical areas, due to its biodegradability, biocompatibility, non-toxicity and versatile chemical and physical properties. Chitosan also presents high antimicrobial activities against a wide variety of pathogenic and spoilage microorganisms. Ultrasonication is a common tool for the preparation and processing of polymer nanoparticles. It is particularly effective in breaking up aggregates and in reducing the size and polydispersity of nanoparticles. High-intensity ultrasonication has the potential to modify chitosan molecular weight and, thus, alter or improve chitosan functional properties. The aim of this study was to evaluate the influence of sonication intensity and time on the changes of commercial chitosan characteristics, such as molecular weight and its potential antibacterial activity against Gram-negative bacteria. The nanoparticles (NPs) were produced from two commercial chitosans, of medium molecular weight (CS-MMW) and low molecular weight (CS-LMW) from Sigma-Aldrich®. These samples (2%) were solubilized in 100 mM sodium acetate pH 4.0, placed on ice and irradiated with an ultrasound SONIC ultrasonic probe (model 750 W), equipped with a 1/2" microtip during 30 min at 4°C. It was used on constant duty cycle and 40% amplitude with 1/1s intervals. The ultrasonic degradation of CS-MMW and CS-LMW were followed up by means of ζ-potential (Brookhaven Instruments, model 90Plus) and dynamic light scattering (DLS) measurements. After sonication, the concentrated samples were diluted 100 times and placed in fluorescence quartz cuvettes (Hellma 111-QS, 10 mm light path). The distributions of the colloidal particles were calculated from the DLS and ζ-potential are measurements taken for the CS-MMW and CS-LMW solutions before and after (CS-MMW30 and CS-LMW30) sonication for 30 min. Regarding the results for the chitosan sample, the major bands can be distinguished centered at Radius hydrodynamic (Rh), showed different distributions for CS-MMW (Rh=690.0 nm, ζ=26.52±2.4), CS-LMW (Rh=607.4 and 2805.4 nm, ζ=24.51±1.29), CS-MMW30 (Rh=201.5 and 1064.1 nm, ζ=24.78±2.4) and CS-LMW30 (Rh=492.5, ζ=26.12±0.85). The minimal inhibitory concentration (MIC) was determined using different chitosan samples concentrations. MIC values were determined against to E. coli (106 cells) harvested from an LB medium (Luria-Bertani BD™) after 18h growth at 37 ºC. Subsequently, the cell suspension was serially diluted in saline solution (0.8% NaCl) and plated on solid LB at 37°C for 18 h. Colony-forming units were counted. The samples showed different MICs against E. coli for CS-LMW (1.5mg), CS-MMW30 (1.5 mg/mL) and CS-LMW30 (1.0 mg/mL). The results demonstrate that the production of nanoparticles by modification of their molecular weight by ultrasonication is simple to be performed and dispense acid solvent addition. Molecular weight modifications are enough to provoke changes in the antimicrobial potential of the nanoparticles produced in this way.

Keywords: antimicrobial agent, chitosan, green production, nanoparticles

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Authors: David Amoah Oduro

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In the ever-evolving terrain of international business, a profound revolution is underway, guided by the swift integration and advancement of disruptive technologies like digital analytics. In today's international business landscape, where competition is fierce, and decisions are data-driven, the essence of this paper lies in offering a tangible roadmap for practitioners. It is a guide that bridges the chasm between theory and actionable insights, helping businesses, investors, and entrepreneurs navigate the complexities of international expansion into sub-Saharan Africa. This practitioner paper distils essential insights, methodologies, and actionable recommendations for businesses seeking to leverage digital analytics in their pursuit of market entry and expansion across the African continent. What sets this paper apart is its unwavering focus on a region ripe with potential: sub-Saharan Africa. The adoption and adaptation of digital analytics are not mere luxuries but essential strategic tools for evaluating countries and entering markets within this dynamic region. With the spotlight firmly fixed on sub-Saharan Africa, the aim is to provide a compelling resource to guide practitioners in their quest to unearth the vast opportunities hidden within sub-Saharan Africa's digital desert. The paper illuminates the pivotal role of digital analytics in providing a data-driven foundation for market entry decisions. It highlights the ability to uncover market trends, consumer behavior, and competitive landscapes. By understanding Africa's incredible diversity, the paper underscores the importance of tailoring market entry strategies to account for unique cultural, economic, and regulatory factors. For practitioners, this paper offers a set of actionable recommendations, including the creation of cross-functional teams, the integration of local expertise, and the cultivation of long-term partnerships to ensure sustainable market entry success. It advocates for a commitment to continuous learning and flexibility in adapting strategies as the African market evolves. This paper represents an invaluable resource for businesses, investors, and entrepreneurs who are keen on unlocking the potential of digital analytics for informed market entry in Africa. It serves as a guiding light, equipping practitioners with the essential tools and insights needed to thrive in this dynamic and diverse continent. With these key insights, methodologies, and recommendations, this paper is a roadmap to prosperous and sustainable market entry in Africa. It is vital for anyone looking to harness the transformational potential of digital analytics to create prosperous and sustainable ventures in a region brimming with promise. In the ever-advancing digital age, this practitioner paper becomes a lodestar, guiding businesses and visionaries toward success amidst the unique challenges and rewards of sub-Saharan Africa's international business landscape.

Keywords: global analytics, digital analytics, sub-Saharan Africa, data analytics

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Authors: Jie Lin, Yiwen Pan, Li Zhang, Zhangyong Xia

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Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of lipids, immune cells, and extracellular matrix in the arterial walls. This pathological process can lead to the formation of plaques that can obstruct blood flow and trigger various cardiovascular diseases such as heart attack and stroke. The underlying molecular mechanisms still remain unclear, although many studies revealed the dysfunction of endothelial cells, recruitment and activation of monocytes and macrophages, and the production of pro-inflammatory cytokines and chemokines in atherosclerosis. This study aimed to identify hub genes involved in the progression of atherosclerosis and to analyze their biological function in silico, thereby enhancing our understanding of the disease’s molecular mechanisms. Through the analysis of microarray data, we examined the gene expression in media and neo-intima from plaques, as well as distant macroscopically intact tissue, across a cohort of 32 hypertensive patients. Initially, 112 differentially expressed genes (DEGs) were identified. Subsequent immune infiltration analysis indicated a predominant presence of 27 immune cell types in the atherosclerosis group, particularly noting an increase in monocytes and macrophages. In the Weighted gene co-expression network analysis (WGCNA), 10 modules with a minimum of 30 genes were defined as key modules, with blue, dark, Oliver green and sky-blue modules being the most significant. These modules corresponded respectively to monocyte, activated B cell, and activated CD4 T cell gene patterns, revealing a strong morphological-genetic correlation. From these three gene patterns (modules morphology), a total of 2509 key genes (Gene Significance >0.2, module membership>0.8) were extracted. Six hub genes (CD36, DPP4, HMOX1, PLA2G7, PLN2, and ACADL) were then identified by intersecting 2509 key genes, 102 DEGs with lipid-related genes from the Genecard database. The bio-functional analysis of six hub genes was estimated by a robust classifier with an area under the curve (AUC) of 0.873 in the ROC plot, indicating excellent efficacy in differentiating between the disease and control group. Moreover, PCA visualization demonstrated clear separation between the groups based on these six hub genes, suggesting their potential utility as classification features in predictive models. Protein-protein interaction (PPI) analysis highlighted DPP4 as the most interconnected gene. Within the constructed key gene-drug network, 462 drugs were predicted, with ursodeoxycholic acid (UDCA) being identified as a potential therapeutic agent for modulating DPP4 expression. In summary, our study identified critical hub genes implicated in the progression of atherosclerosis through comprehensive bioinformatic analyses. These findings not only advance our understanding of the disease but also pave the way for applying similar analytical frameworks and predictive models to other diseases, thereby broadening the potential for clinical applications and therapeutic discoveries.

Keywords: atherosclerosis, hub genes, drug prediction, bioinformatics

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

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

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

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Authors: Rikke Lybæk, Tyge Kjær

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The Baltic Sea is suffering from nitrogen and phosphorus pollution, which causes eutrophication of the maritime environment and hence threatens the biodiversity of the Baltic Sea area. The intensified quantity of nutrients in the water has created challenges with the growth of seaweed being discarded on beaches around the sea. The cast seaweed has led to odor problems hampering the use of beach areas around the Bay of Køge in Denmark. This is the case in, e.g., Solrød Municipality, where recreational activities have been disrupted when cast seaweed pile up on the beach. Initiatives have, however, been introduced within the municipality to remove the cast seaweed from the beach and utilize it for renewable energy production at the nearby Solrød Biogas Plant, thus being co-digested with animal manure for power and heat production. This paper investigates which type of technology application’s have been applied in the effort to optimize the collection of cast seaweed, and will further reveal, how the seaweed has been pre-treated at the biogas plant to be utilized for energy production the most efficient, hereunder the challenges connected with the content of sand. Heavy metal contents in the seaweed and how it is managed will also be addressed, which is vital as the digestate is utilized as soil fertilizer on nearby farms. Finally, the paper will outline the energy production scheme connected to the use of seaweed as feedstock for biogas production, as well as the amount of nitrogen-rich fertilizer produced. The theoretical approach adopted in the paper relies on the thinking of Circular Bio-Economy, where biological materials are cascaded and re-circulated etc., to increase and extend their value and usability. The data for this research is collected as part of the EU Interreg project “Cluster On Anaerobic digestion, environmental Services, and nuTrients removAL” (COASTAL Biogas), 2014-2020. Data gathering consists of, e.g., interviews with relevant stakeholders connected to seaweed collection and operation of the biogas plant in Solrød Municipality. It further entails studies of progress and evaluation reports from the municipality, analysis of seaweed digestion results from scholars connected to the research, as well as studies of scientific literature to supplement the above. Besides this, observations and photo documentation have been applied in the field. This paper concludes, among others, that the seaweed harvester technology currently adopted is functional in the maritime environment close to the beachfront but inadequate in collecting seaweed directly on the beach. New technology hence needs to be developed to increase the efficiency of seaweed collection. It is further concluded that the amount of sand transported to Solrød Biogas Plant with the seaweed continues to pose challenges. The seaweed is pre-treated for sand in a receiving tank with a strong stirrer, washing off the sand, which ends at the bottom of the tank where collected. The seaweed is then chopped by a macerator and mixed with the other feedstock. The wear down of the receiving tank stirrer and the chopper are, however, significant, and new methods should be adopted.

Keywords: biogas, circular bio-economy, Denmark, maritime technology, cast seaweed, solrød municipality

Procedia PDF Downloads 258

Authors: Nicola Milani, Carla Bellissimo, Davide Pogliana, Davide Panzin, Luca Garlaschelli, Giulia Facchinetti, Claudia Casson, Luca Marazzina, Andrea Sartori, Simone Rivaroli, Jeff Konin

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The patient is a 26 year-old male amateur football player from Milan, Italy; (81kg; 185cm; BMI 23.6 kg/m²). He sustained a non-contact anterior cruciate ligament tear to his right knee in June 2021. In September 2021, his right knee ligament was reconstructed using a semitendinosus graft. The injury occurred during a football match on natural grass with typical shoes on a warm day (32 degrees celsius). Playing as a defender he sustained the injury during a change of direction, where the foot was fixated on the grass. He felt pain and was unable to continue playing the match. The surgeon approved his rehabilitation to begin two weeks post-operative. The initial physiotherapist assessment determined performing two training sessions per day within the first three months. In the first three weeks, the pain was 4/10 on Numerical Rating Scale (NRS), no swelling, a range of motion was 0-110°, with difficulty fully extending his knee and minimal quadriceps activation. Crutches were discontinued at four weeks with improved walking. Active exercise, electrostimulator, physical therapy, massages, osteopathy, and passive motion were initiated. At week 6, he completed his first functional movement screen; the score was 16/21 with no pain and no swelling. At week 8, the isokinetic test showed a 23% differential deficit between the two legs in maximum strength (at 90°/s). At week 10, he improved to 15% of injury-induced deficit which suggested he was ready to start running. At week 12, the athlete sustained his first threshold test. At week 16, he performed his first return to sports movement assessment, which revealed a 10% stronger difference between the legs. At week 16, he had his second threshold test. At week 17, his first on-field test revealed a 5% differential deficit between the two legs in the hop test. At week 18, isokinetic test demonstrates that the uninjured leg was 7% stronger than the recovering leg in maximum strength (at 90°/s). At week 20, his second on-field test revealed a 2% difference in hop test; at week 21, his third isokinetic test demonstrated a difference of 5% in maximum strength (at 90°/s). At week 21, he performed his second return to sports movement assessment which revealed a 2% difference between the limbs. Since it was the end of the championship, the team asked him to partake in the playoffs; moreover the player was very motivated to participate in the playoffs also because he was the captain of the team. Together with the player and the team, we decided to let him play even though we were aware of a heightened risk of injury than what is reported in the literature because of two factors: biological recovery times and the results of the tests we performed. In the decision making process about the athlete’s recovery time, it is important to balance the information available from the literature with the desires of the patient to avoid frustration.

Keywords: ACL, football, rehabilitation, return to play

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Authors: Madina Tynybayeva, Sanya Zhumazhanova, Saltanat Kozhakhmetova, Merey Mussabayeva

Abstract:

One of the leading indicators of the education quality is the level of students’ educational achievements. The processes of modernization of Kazakhstani education system have predetermined the need to improve the national system by assessing the quality of education. The results of assessment greatly contribute to addressing questions about the current state of the educational system in the country. The monitoring of students’ educational achievements (MEAS) is the systematic measurement of the quality of education for compliance with the state obligatory standard of Kazakhstan. This systematic measurement is independent of educational organizations and approved by the order of the Minister of Education and Scienceof Kazakhstan. The MEAS was conducted in the regions of Kazakhstanfor the first time in 2022 by the National Testing Centre. The measurement does not have legal consequences either for students or for educational organizations. Students’ achievements were measured in three subject areas: reading, mathematics and science literacy. MEAS was held for the first time in April this year, 105 thousand students from 1436 schools of Kazakhstan took part in the testing. The monitoring was accompanied by a survey of students, teachers, and school leaders. The goal is to identify which contextual factors affect learning outcomes. The testing was carried out in a computer format. The test tasks of MEAS are ranked according to the three levels of difficulty: basic, medium, and high. Fourth graders are asked to complete 30 closed-type tasks. The average score of the results is 21 points out of 30, which means 70% of tasks were successfully completed. The total number of test tasks for 9th grade students – 75 questions. The results of ninth graders are comparatively lower, the success rate of completing tasks is 63%. MEAS participants did not reveal a statistically significant gap in results in terms of the language of instruction, territorial status, and type of school. The trend of reducing the gap in these indicators is also noted in the framework of recent international studies conducted across the country, in particular PISA for schools in Kazakhstan. However, there is a regional gap in MOES performance. The difference in the values of the indicators of the highest and lowest scores of the regions was 11% of the success of completing tasks in the 4th grade, 14% in the 9thgrade. The results of the 4th grade students in reading, mathematics, and science literacy are: 71.5%, 70%, and 66.9%, respectively. The results of ninth-graders in reading, mathematics, and science literacy are 69.6%, 54%, and 60.8%, respectively. From the surveys, it was revealed that the educational achievements of students are considerably influenced by such factors as the subject competences of teachers, as well as the school climate and motivation of students. Thus, the results of MEAS indicate the need for an integrated approach to improving the quality of education. In particular, the combination of improving the content of curricula and textbooks, internal and external assessment of the educational achievements of students, educational programs of pedagogical specialties, and advanced training courses is required.

Keywords: assessment, secondary school, monitoring, functional literacy, kazakhstan

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Authors: N. Manjunatha, K. T. Rangswamy, N. Nagaraju, H. A. Prameela, P. Rudraswamy, M. Krishnareddy

Abstract:

The identification of virus in cowpea especially potyviruses is confusing. Even though there are several studies on viruses causing diseases in cowpea, difficult to distinguish based on symptoms and serological detection. The differentiation of potyviruses considering as a constraint, the present study is initiated for molecular characterization, host plant resistance and epidemiology of the BCMV infecting cowpea. The etiological agent causing cowpea mosaic was identified as Bean Common Mosaic Virus (BCMV) on the basis of RT-PCR and electron microscopy. An approximately 750bp PCR product corresponding to coat protein (CP) region of the virus and the presence of long flexuous filamentous particles measuring about 952 nm in size typical to genus potyvirus were observed under electron microscope. The characterized virus isolate genome had 10054 nucleotides, excluding the 3’ terminal poly (A) tail. Comparison of polyprotein of the virus with other potyviruses showed similar genome organization with 9 cleavage sites resulted in 10 functional proteins. The pairwise sequence comparison of individual genes, P1 showed most divergent, but CP gene was less divergent at nucleotide and amino acid level. A phylogenetic tree constructed based on multiple sequence alignments of the polyprotein nucleotide and amino acid sequences of cowpea BCMV and potyviruses showed virus is closely related to BCMV-HB. Whereas, Soybean variant of china (KJ807806) and NL1 isolate (AY112735) showed 93.8 % (5’UTR) and 94.9 % (3’UTR) homology respectively with other BCMV isolates. This virus transmitted to different leguminous plant species and produced systemic symptoms under greenhouse conditions. Out of 100 cowpea genotypes screened, three genotypes viz., IC 8966, V 5 and IC 202806 showed immune reaction in both field and greenhouse conditions. Single marker analysis (SMA) was revealed out of 4 SSR markers linked to BCMV resistance, M135 marker explains 28.2 % of phenotypic variation (R2) and Polymorphic information content (PIC) value of these markers was ranged from 0.23 to 0.37. The correlation and regression analysis showed rainfall, and minimum temperature had significant negative impact and strong relationship with aphid population, whereas weak correlation was observed with disease incidence. Path coefficient analysis revealed most of the weather parameters exerted their indirect contributions to the aphid population and disease incidence except minimum temperature. This study helps to identify specific gaps in knowledge for researchers who may wish to further analyse the science behind complex interactions between vector-virus and host in relation to the environment. The resistant genotypes identified are could be effectively used in resistance breeding programme.

Keywords: cowpea, epidemiology, genotypes, virus

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Authors: Poojan Gharat, Haridas Pal, Sharmistha Dutta Choudhury

Abstract:

Tuning photophysical properties of guest dyes through host-guest interactions involving macrocyclic hosts are the attractive research areas since past few decades, as these changes can directly be implemented in chemical sensing, molecular recognition, fluorescence imaging and dye laser applications. Excited state intramolecular proton transfer (ESIPT) is an intramolecular prototautomerization process display by some specific dyes. The process is quite amenable to tunability by the presence of different macrocyclic hosts. The present study explores the interesting effect of p-sulfonatocalix[n]arene (SCXn) and cyclodextrin (CD) hosts on the excited-state prototautomeric equilibrium of Chrysazine (CZ), a model antitumour drug. CZ exists exclusively in its normal form (N) in the ground state. However, in the excited state, the excited N* form undergoes ESIPT along with its pre-existing intramolecular hydrogen bonds, giving the excited state prototautomer (T*). Accordingly, CZ shows a single absorption band due to N form, but two emission bands due to N* and T* forms. Facile prototautomerization of CZ is considerably inhibited when the dye gets bound to SCXn hosts. However, in spite of lower binding affinity, the inhibition is more profound with SCX6 host as compared to SCX4 host. For CD-CZ system, while prototautomerization process is hindered by the presence of β-CD, it remains unaffected in the presence of γCD. Reduction in the prototautomerization process of CZ by SCXn and βCD hosts is unusual, because T* form is less dipolar in nature than the N*, hence binding of CZ within relatively hydrophobic hosts cavities should have enhanced the prototautomerization process. At the same time, considering the similar chemical nature of two CD hosts, their effect on prototautomerization process of CZ would have also been similar. The atypical effects on the prototautomerization process of CZ by the studied hosts are suggested to arise due to the partial inclusion or external binding of CZ with the hosts. As a result, there is a strong possibility of intermolecular H-bonding interaction between CZ dye and the functional groups present at the portals of SCXn and βCD hosts. Formation of these intermolecular H-bonds effectively causes the pre-existing intramolecular H-bonding network within CZ molecule to become weak, and this consequently reduces the prototautomerization process for the dye. Our results suggest that rather than the binding affinity between the dye and host, it is the orientation of CZ in the case of SCXn-CZ complexes and the binding stoichiometry in the case of CD-CZ complexes that play the predominant role in influencing the prototautomeric equilibrium of the dye CZ. In the case of SCXn-CZ complexes, the results obtained through experimental findings are well supported by quantum chemical calculations. Similarly for CD-CZ systems, binding stoichiometries obtained through geometry optimization studies on the complexes between CZ and CD hosts correlate nicely with the experimental results. Formation of βCD-CZ complexes with 1:1 stoichiometry while formation of γCD-CZ complexes with 1:1, 1:2 and 2:2 stoichiometries are revealed from geometry optimization studies and these results are in good accordance with the observed effects by the βCD and γCD hosts on the ESIPT process of CZ dye.

Keywords: intermolecular proton transfer, macrocyclic hosts, quantum chemical studies, photophysical studies

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Authors: Jun Wang, Chenglei Diao, Emanuele Pagone, Jialuo Ding, Stewart Williams

Abstract:

As a crucial component in aircraft, landing gear systems are responsible for supporting the plane during parking, taxiing, takeoff, and landing. Given the need for high load-bearing capacity over extended periods, 300M ultra-high strength steel (UHSS) is often the material of choice for crafting these systems due to its exceptional strength, toughness, and fatigue resistance. In the quest for cost-effective and sustainable manufacturing solutions, Wire Arc Additive Manufacturing (WAAM) emerges as a promising alternative for fabricating 300M UHSS landing gears. This is due to its advantages in near-net-shape forming of large components, cost-efficiency, and reduced lead times. Cranfield University has conducted an extensive preliminary study on WAAM 300M UHSS, covering feature deposition, interface analysis, and post-heat treatment. Both Gas Metal Arc (GMA) and Plasma Transferred Arc (PTA)-based WAAM methods were explored, revealing their feasibility for defect-free manufacturing. However, as-deposited 300M features showed lower strength but higher ductility compared to their forged counterparts. Subsequent post-heat treatments were effective in normalising the microstructure and mechanical properties, meeting qualification standards. A 300M UHSS landing gear demonstrator was successfully created using PTA-based WAAM, showcasing the method's precision and cost-effectiveness. The demonstrator, measuring Ф200mm x 700mm, was completed in 16 hours, using 7 kg of material at a deposition rate of 1.3kg/hr. This resulted in a significant reduction in the Buy-to-Fly (BTF) ratio compared to traditional manufacturing methods, further validating WAAM's potential for this application. A "cradle-to-gate" environmental impact assessment, which considers the cumulative effects from raw material extraction to customer shipment, has revealed promising outcomes. Utilising Wire Arc Additive Manufacturing (WAAM) for landing gear components significantly reduces the need for raw material extraction and refinement compared to traditional subtractive methods. This, in turn, lessens the burden on subsequent manufacturing processes, including heat treatment, machining, and transportation. Our estimates indicate that the carbon footprint of the component could be halved when switching from traditional machining to WAAM. Similar reductions are observed in embodied energy consumption and other environmental impact indicators, such as emissions to air, water, and land. Additionally, WAAM offers the unique advantage of part repair by redepositing only the necessary material, a capability not available through conventional methods. Our research shows that WAAM-based repairs can drastically reduce environmental impact, even when accounting for additional transportation for repairs. Consequently, WAAM emerges as a pivotal technology for reducing environmental impact in manufacturing, aiding the industry in its crucial and ambitious journey towards Net Zero. This study paves the way for transformative benefits across the aerospace industry, as we integrate manufacturing into a hybrid solution that offers substantial savings and access to more sustainable technologies for critical component production.

Keywords: WAAM, aircraft landing gear, microstructure, mechanical performance, life cycle assessment

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Authors: Reham Khalaf-Nazzal, Imad Dweikat, Paula Gimenez, Iker Oyenarte, Alfonso Martinez-Cruz, Domonik Muller

Abstract:

Metal cation transport mediator (CNNM) gene family comprises 4 isoforms that are expressed in various human tissues. Structurally, CNNMs are complex proteins that contain an extracellular N-terminal domain preceding a DUF21 transmembrane domain, a ‘Bateman module’ and a C-terminal cNMP-binding domain. Mutations in CNNM2 cause familial dominant hypomagnesaemia. Growing evidence highlights the role of CNNM2 in neurodevelopment. Mutations in CNNM2 have been implicated in epilepsy, intellectual disability, schizophrenia, and others. In the present study, we aim to elucidate the function of CNNM2 in the developing brain. Thus, we present the genetic origin of symptoms in two family cohorts. In the first family, three siblings of a consanguineous Palestinian family in which parents are first cousins, and consanguinity ran over several generations, presented a varying degree of intellectual disability, cone-rod dystrophy, and autism spectrum disorder. Exome sequencing and segregation analysis revealed the presence of homozygous pathogenic mutation in the CNNM2 gene, the parents were heterozygous for that gene mutation. Magnesium blood levels were normal in the three children and their parents in several measurements. They had no symptoms of hypomagnesemia. The CNNM2 mutation in this family was found to locate in the CBS1 domain of the CNNM2 protein. The crystal structure of the mutated CNNM2 protein was not significantly different from the wild-type protein, and the binding of AMP or MgATP was not dramatically affected. This suggests that the CBS1 domain could be involved in pure neurodevelopmental functions independent of its magnesium-handling role, and this mutation could have affected a protein partner binding or other functions in this protein. In the second family, another autosomal dominant CNNM2 mutation was found to run in a large family with multiple individuals over three generations. All affected family members had hypomagnesemia and hypermagnesuria. Oral supplementation of magnesium did not increase the levels of magnesium in serum significantly. Some affected members of this family have defects in fine motor skills such as dyslexia and dyslalia. The detected mutation is located in the N-terminal part, which contains a signal peptide thought to be involved in the sorting and routing of the protein. In this project, we describe heterogenous clinical phenotypes related to CNNM2 mutations and protein functions. In the first family, and up to the authors’ knowledge, we report for the first time the involvement of CNNM2 in retinal photoreceptor development and function. In addition, we report the presence of a neurophenotype independent of magnesium status related to the CNNM2 protein mutation. Taking into account the different modes of inheritance and the different positions of the mutations within CNNM2 and its different structural and functional domains, it is likely that CNNM2 might be involved in a wide spectrum of neuropsychiatric comorbidities with considerable varying phenotypes.

Keywords: magnesium transport, autosomal recessive, autism, neurodevelopment, CBS domain

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