Search results for: mechanisms of plant defense
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 6110

Search results for: mechanisms of plant defense

530 Chinese Acupuncture: A Potential Treatment for Autism Rat Model via Improving Synaptic Function

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

Abstract:

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

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

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529 Real-Space Mapping of Surface Trap States in Cigse Nanocrystals Using 4D Electron Microscopy

Authors: Riya Bose, Ashok Bera, Manas R. Parida, Anirudhha Adhikari, Basamat S. Shaheen, Erkki Alarousu, Jingya Sun, Tom Wu, Osman M. Bakr, Omar F. Mohammed

Abstract:

This work reports visualization of charge carrier dynamics on the surface of copper indium gallium selenide (CIGSe) nanocrystals in real space and time using four-dimensional scanning ultrafast electron microscopy (4D S-UEM) and correlates it with the optoelectronic properties of the nanocrystals. The surface of the nanocrystals plays a key role in controlling their applicability for light emitting and light harvesting purposes. Typically for quaternary systems like CIGSe, which have many desirable attributes to be used for optoelectronic applications, relative abundance of surface trap states acting as non-radiative recombination centre for charge carriers remains as a major bottleneck preventing further advancements and commercial exploitation of these nanocrystals devices. Though ultrafast spectroscopic techniques allow determining the presence of picosecond carrier trapping channels, because of relative larger penetration depth of the laser beam, only information mainly from the bulk of the nanocrystals is obtained. Selective mapping of such ultrafast dynamical processes on the surfaces of nanocrystals remains as a key challenge, so far out of reach of purely optical probing time-resolved laser techniques. In S-UEM, the optical pulse generated from a femtosecond (fs) laser system is used to generate electron packets from the tip of the scanning electron microscope, instead of the continuous electron beam used in the conventional setup. This pulse is synchronized with another optical excitation pulse that initiates carrier dynamics in the sample. The principle of S-UEM is to detect the secondary electrons (SEs) generated in the sample, which is emitted from the first few nanometers of the top surface. Constructed at different time delays between the optical and electron pulses, these SE images give direct and precise information about the carrier dynamics on the surface of the material of interest. In this work, we report selective mapping of surface dynamics in real space and time of CIGSe nanocrystals applying 4D S-UEM. We show that the trap states can be considerably passivated by ZnS shelling of the nanocrystals, and the carrier dynamics can be significantly slowed down. We also compared and discussed the S-UEM kinetics with the carrier dynamics obtained from conventional ultrafast time-resolved techniques. Additionally, a direct effect of the state trap removal can be observed in the enhanced photoresponse of the nanocrystals after shelling. Direct observation of surface dynamics will not only provide a profound understanding of the photo-physical mechanisms on nanocrystals’ surfaces but also enable to unlock their full potential for light emitting and harvesting applications.

Keywords: 4D scanning ultrafast microscopy, charge carrier dynamics, nanocrystals, optoelectronics, surface passivation, trap states

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528 Ways to Prevent Increased Wear of the Drive Box Parts and the Central Drive of the Civil Aviation Turbo Engine Based on Tribology

Authors: Liudmila Shabalinskaya, Victor Golovanov, Liudmila Milinis, Sergey Loponos, Alexander Maslov, D. O. Frolov

Abstract:

The work is devoted to the rapid laboratory diagnosis of the condition of aircraft friction units, based on the application of the nondestructive testing method by analyzing the parameters of wear particles, or tribodiagnostics. The most important task of tribodiagnostics is to develop recommendations for the selection of more advanced designs, materials and lubricants based on data on wear processes for increasing the life and ensuring the safety of the operation of machines and mechanisms. The object of tribodiagnostics in this work are the tooth gears of the central drive and the gearboxes of the gas turbine engine of the civil aviation PS-90A type, in which rolling friction and sliding friction with slip occur. The main criterion for evaluating the technical state of lubricated friction units of a gas turbine engine is the intensity and rate of wear of the friction surfaces of the friction unit parts. When the engine is running, oil samples are taken and the state of the friction surfaces is evaluated according to the parameters of the wear particles contained in the oil sample, which carry important and detailed information about the wear processes in the engine transmission units. The parameters carrying this information include the concentration of wear particles and metals in the oil, the dispersion composition, the shape, the size ratio and the number of particles, the state of their surfaces, the presence in the oil of various mechanical impurities of non-metallic origin. Such a morphological analysis of wear particles has been introduced into the order of monitoring the status and diagnostics of various aircraft engines, including a gas turbine engine, since the type of wear characteristic of the central drive and the drive box is surface fatigue wear and the beginning of its development, accompanied by the formation of microcracks, leads to the formation of spherical, up to 10 μm in size, and in the aftermath of flocculent particles measuring 20-200 μm in size. Tribodiagnostics using the morphological analysis of wear particles includes the following techniques: ferrography, filtering, and computer analysis of the classification and counting of wear particles. Based on the analysis of several series of oil samples taken from the drive box of the engine during their operating time, a study was carried out of the processes of wear kinetics. Based on the results of the study and comparing the series of criteria for tribodiagnostics, wear state ratings and statistics of the results of morphological analysis, norms for the normal operating regime were developed. The study allowed to develop levels of wear state for friction surfaces of gearing and a 10-point rating system for estimating the likelihood of the occurrence of an increased wear mode and, accordingly, prevention of engine failures in flight.

Keywords: aviation, box of drives, morphological analysis, tribodiagnostics, tribology, ferrography, filtering, wear particle

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527 Usage of Cyanobacteria in Battery: Saving Money, Enhancing the Storage Capacity, Making Portable, and Supporting the Ecology

Authors: Saddam Husain Dhobi, Bikrant Karki

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The main objective of this paper is save money, balance ecosystem of the terrestrial organism, control global warming, and enhancing the storage capacity of the battery with requiring weight and thinness by using Cyanobacteria in the battery. To fulfill this purpose of paper we can use different methods: Analysis, Biological, Chemistry, theoretical and Physics with some engineering design. Using this different method, we can produce the special type of battery that has the long life, high storage capacity, and clean environment, save money so on and by using the byproduct of Cyanobacteria i.e. glucose. Cyanobacteria are a special type of bacteria that produces different types of extracellular glucoses and oxygen with the help of little sunlight, water, and carbon dioxide and can survive in freshwater, marine and in the land as well. In this process, O₂ is more in the comparison to plant due to rapid growth rate of Cyanobacteria. The required materials are easily available in this process to produce glucose with the help of Cyanobacteria. Since CO₂, is greenhouse gas that causes the global warming? We can utilize this gas and save our ecological balance and the byproduct (glucose) C₆H₁₂O₆ can be utilized for raw material for the battery where as O₂ escape is utilized by living organism. The glucose produce by Cyanobateria goes on Krebs's Cycle or Citric Acid Cycle, in which glucose is complete, oxidizes and all the available energy from glucose molecule has been release in the form of electron and proton as energy. If we use a suitable anodes and cathodes, we can capture these electrons and protons to produce require electricity current with the help of byproduct of Cyanobacteria. According to "Virginia Tech Bio-battery" and "Sony" 13 enzymes and the air is used to produce nearly 24 electrons from a single glucose unit. In this output power of 0.8 mW/cm, current density of 6 mA/cm, and energy storage density of 596 Ah/kg. This last figure is impressive, at roughly 10 times the energy density of the lithium-ion batteries in your mobile devices. When we use Cyanobacteria in battery, we are able to reduce Carbon dioxide, Stop global warming, and enhancing the storage capacity of battery more than 10 times that of lithium battery, saving money, balancing ecology. In this way, we can produce energy from the Cyanobacteria and use it in battery for different benefits. In addition, due to the mass, size and easy cultivation, they are better to maintain the size of battery. Hence, we can use Cyanobacteria for the battery having suitable size, enhancing the storing capacity of battery, helping the environment, portability and so on.

Keywords: anode, byproduct, cathode, cyanobacteri, glucose, storage capacity

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526 Examining the Relationship Between Green Procurement Practices and Firm’s Performance in Ghana

Authors: Alexander Otchere Fianko, Clement Yeboah, Evans Oteng

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Prior research concludes that Green Procurement Practices positively drive Organisational Performance. Nonetheless, the nexus and conditions under which Green Procurement Practices contribute to a Firm’s Performance are less understood. The purpose of this quantitative relational study was to examine the relationship between Green Procurement Practices and 500 Firms’ Performances in Ghana. The researchers further seek to draw insights from the resource-based view to conceptualize Green Procurement Practices and Environmental Commitment as resource capabilities to enhance Firm Performance. The researchers used insights from the contingent resource-based view to examine Green Leadership Orientation conditions under which Green Procurement Practices contribute to Firm Performance through Environmental Commitment Capabilities. The study’s conceptual framework was tested on primary data from some firms in the Ghanaian market. PROCESS Macro was used to test the study’s hypotheses. Beyond that, Environmental Commitment Capabilities mediated the association between Green Procurement Practices and the Firm’s Performance. The study further seeks to find out whether Green Leadership Orientation positively moderates the indirect relationship between Green Procurement Practices and Firm Performance through Environmental Commitment Capabilities. While conventional wisdom suggests that improved Green Procurement Practices help improve a Firm’s Performance, this study tested this presumed relationship between Green Procurement Practices and Firm Performance and provides theoretical arguments and empirical evidence to justify how Environmental Commitment Capabilities uniquely and in synergy with Green Leadership Orientation transform this relationship. The study results indicated a positive correlation between Green Procurement Practices and Firm Performance. This result suggests that firms that prioritize environmental sustainability and demonstrate a strong commitment to environmentally responsible practices tend to experience better overall performance. This includes financial gains, operational efficiency, enhanced reputation, and improved relationships with stakeholders. The study's findings inform policy formulation in Ghana related to environmental regulations, incentives, and support mechanisms. Policymakers can use the insights to design policies that encourage and reward firms for their Green Procurement Practices, thereby fostering a more sustainable and environmentally responsible business environment. The findings from such research can influence the design and development of educational programs in Ghana, specifically in fields related to sustainability, environmental management, and corporate social responsibility (CSR). Institutions may consider integrating environmental and sustainability topics into their business and management courses to create awareness and promote responsible practices among future business professionals. Also, the study results can also promote the adoption of environmental accounting practices in Ghana. By recognizing and measuring the environmental impacts and costs associated with business activities, firms can better understand the financial implications of their Green Procurement Practices and develop strategies for improved performance.

Keywords: environmental commitment, firm’s performance, green procurement practice, green leadership orientation

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525 Assessment of OTA Contamination in Rice from Fungal Growth Alterations in a Scenario of Climate Changes

Authors: Carolina S. Monteiro, Eugénia Pinto, Miguel A. Faria, Sara C. Cunha

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Rice (Oryza sativa) production plays a vital role in reducing hunger and poverty and assumes particular importance in low-income and developing countries. Rice is a sensitive plant, and production occurs strictly where suitable temperature and water conditions are found. Climatic changes are likely to affect worldwide, and some models have predicted increased temperatures, variations in atmospheric CO₂ concentrations and modification in precipitation patterns. Therefore, the ongoing climatic changes threaten rice production by increasing biotic and abiotic stress factors, and crops will grow in different environmental conditions in the following years. Around the world, the effects will be regional and can be detrimental or advantageous depending on the region. Mediterranean zones have been identified as possible hot spots, where dramatic temperature changes, modifications of CO₂ levels, and rainfall patterns are predicted. The actual estimated atmospheric CO₂ concentration is around 400 ppm, and it is predicted that it can reach up to 1000–1200 ppm, which can lead to a temperature increase of 2–4 °C. Alongside, rainfall patterns are also expected to change, with more extreme wet/dry episodes taking place. As a result, it could increase the migration of pathogens, and a shift in the occurrence of mycotoxins, concerning their types and concentrations, is expected. Mycotoxigenic spoilage fungi can colonize the crops and be present in all rice food chain supplies, especially Penicillium species, mainly resulting in ochratoxin A (OTA) contamination. In this scenario, the objectives of the present study are evaluating the effect of temperature (20 vs. 25 °C), CO₂ (400 vs. 1000 ppm), and water stress (0.93 vs 0.95 water activity) on growth and OTA production by a Penicillium nordicum strain in vitro on rice-based media and when colonizing layers of raw rice. Results demonstrate the effect of temperature, CO₂ and drought on the OTA production in a rice-based environment, thus contributing to the development of mycotoxins predictive models in climate change scenarios. As a result, improving mycotoxins' surveillance and monitoring systems, whose occurrence can be more frequent due to climatic changes, seems relevant and necessary. The development of prediction models for hazard contaminants presents in foods highly sensitive to climatic changes, such as mycotoxins, in the highly probable new agricultural scenarios is of paramount importance.

Keywords: climate changes, ochratoxin A, penicillium, rice

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524 Consensual A-Monogamous Relationships: Challenges and Ways of Coping

Authors: Tal Braverman Uriel, Tal Litvak Hirsch

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Background and Objectives: Little or only partial emphasis has been placed on exploring the complexity of consensual non-monogamous relationships. The term "polyamory" refers to consensual non-monogamy, and it is defined as having emotional and/or sexual relations simultaneously with two or more people, the consent and knowledge of all the partners concerned. Managing multiple romantic relationships with different people evokes more emotions, leads to more emotional conflicts arising from different interests, and demands practical strategies. An individual's transition from a monogamous lifestyle to a consensual non-monogamous lifestyle yields new challenges, accompanied by stress, uncertainty, and question marks, as do other life-changing events, such as divorce or transition to parenthood. The study examines both the process of transition and adaptation to a consensually non-monogamous relationship, as well as the coping mechanism involved in the daily conduct of this lifestyle. The research focuses on understanding the consequences, challenges, and coping methods from a personal, marital, and familial point of view and focuses on 40 middle-aged individuals (20 men and 20 women ages 40-60). The research sheds light on a way of life that has not been previously studied in Israel and is still considered unacceptable. Theories of crisis (e.g., as Folkman and Lazarus) were applied, and as a result, a deeper understanding of the subject was reached, all while focusing on multiple aspects of dealing with stress. The basic research question examines the consequences of entering a polyamorous life from a personal point of view as an individual, partner, and parent and the ways of coping with these consequences. Method: The research is conducted with a narrative qualitative approach in the interpretive paradigm, including semi-structured in-depth interviews. The method of analysis is thematic. Results: The findings indicate that in most cases, an individual's motivation to open the relationship is mainly a longing for better sexuality and for an added layer of excitement to their lives. Most of the interviewees were assisted by their spouses in the process, as well as by social networks and podcasts on the subject. Some of them therapeutic professionals from the field are helpful. It also clearly emerged that among those who experienced acute emotional crises with the primary partner or painful separations from secondary partners, all believed polyamory to be the adequate way of life for them. Finally, a key resource for managing tension and stress is the ability to share and communicate with the primary partner. Conclusions: The study points to the challenges and benefits of a non-monogamous lifestyle as well as the use of coping mechanisms and resources that are consistent with the existing theory and research in the field in the context of life changes. The study indicates the need to expand the research canvas in the future in the context of parenting and the consequences for children.

Keywords: a-monogamy, consent, family, stress, tension

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523 Bioefficiency of Cinnamomum verum Loaded Niosomes and Its Microbicidal and Mosquito Larvicidal Activity against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus

Authors: Aasaithambi Kalaiselvi, Michael Gabriel Paulraj, Ekambaram Nakkeeran

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Emergences of mosquito vector-borne diseases are considered as a perpetual problem globally in tropical countries. The outbreak of several diseases such as chikungunya, zika virus infection and dengue fever has created a massive threat towards the living population. Frequent usage of synthetic insecticides like Dichloro Diphenyl Trichloroethane (DDT) eventually had its adverse harmful effects on humans as well as the environment. Since there are no perennial vaccines, prevention, treatment or drugs available for these pathogenic vectors, WHO is more concerned in eradicating their breeding sites effectively without any side effects on humans and environment by approaching plant-derived natural eco-friendly bio-insecticides. The aim of this study is to investigate the larvicidal potency of Cinnamomum verum essential oil (CEO) loaded niosomes. Cholesterol and surfactant variants of Span 20, 60 and 80 were used in synthesizing CEO loaded niosomes using Transmembrane pH gradient method. The synthesized CEO loaded niosomes were characterized by Zeta potential, particle size, Fourier Transform Infrared Spectroscopy (FT-IR), GC-MS and SEM analysis to evaluate charge, size, functional properties, the composition of secondary metabolites and morphology. The Z-average size of the formed niosomes was 1870.84 nm and had good stability with zeta potential -85.3 meV. The entrapment efficiency of the CEO loaded niosomes was determined by UV-Visible Spectrophotometry. The bio-potency of CEO loaded niosomes was treated and assessed against gram-positive (Bacillus subtilis) and gram-negative (Escherichia coli) bacteria and fungi (Aspergillus fumigatus and Candida albicans) at various concentrations. The larvicidal activity was evaluated against II to IV instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus at various concentrations for 24 h. The mortality rate of LC₅₀ and LC₉₀ values were calculated. The results exhibited that CEO loaded niosomes have greater efficiency against mosquito larvicidal activity. The results suggest that niosomes could be used in various applications of biotechnology and drug delivery systems with greater stability by altering the drug of interest.

Keywords: Cinnamomum verum, niosomes, entrapment efficiency, bactericidal and fungicidal, mosquito larvicidal activity

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522 Climate Change Impact on Water Resources Management in Remote Islands Using Hybrid Renewable Energy Systems

Authors: Elissavet Feloni, Ioannis Kourtis, Konstantinos Kotsifakis, Evangelos Baltas

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Water inadequacy in small dry islands scattered in the Aegean Sea (Greece) is a major problem regarding Water Resources Management (WRM), especially during the summer period due to tourism. In the present work, various WRM schemes are designed and presented. The WRM schemes take into account current infrastructure and include Rainwater Harvesting tanks and Reverse Osmosis Desalination Units. The energy requirements are covered mainly by wind turbines and/or a seawater pumped storage system. Sizing is based on the available data for population and tourism per island, after taking into account a slight increase in the population (up to 1.5% per year), and it guarantees at least 80% reliability for the energy supply and 99.9% for potable water. Evaluation of scenarios is carried out from a financial perspective, after calculating the Life Cycle Cost (LCC) of each investment for a lifespan of 30 years. The wind-powered desalination plant was found to be the most cost-effective practice, from an economic point of view. Finally, in order to estimate the Climate Change (CC) impact, six different CC scenarios were investigated. The corresponding rate of on-grid versus off-grid energy required for ensuring the targeted reliability for the zero and each climatic scenario was investigated per island. The results revealed that under CC the grid-on energy required would increase and as a result, the reduction in wind turbines and seawater pumped storage systems’ reliability will be in the range of 4 to 44%. However, the range of this percentage change does not exceed 22% per island for all examined CC scenarios. Overall, CC is proposed to be incorporated into the design process for WRM-related projects. Acknowledgements: This research is co-financed by Greece and the European Union (European Social Fund - ESF) through the Operational Program «Human Resources Development, Education and Lifelong Learning 2014-2020» in the context of the project “Development of a combined rain harvesting and renewable energy-based system for covering domestic and agricultural water requirements in small dry Greek Islands” (MIS 5004775).

Keywords: small dry islands, water resources management, climate change, desalination, RES, seawater pumped storage system, rainwater harvesting

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521 Phenotypic Diversity of the Tomato Germplasm from the Lazio Region in Central Italy, with a Case Study on Molecular Distinctiveness

Authors: Barbara Farinon, Maurizio E. Picarella, Lorenzo Mancini, Andrea Mazzucato

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Italy is notoriously a secondary center of diversification for cultivated tomatoes (Solanum lycopersicum L.). The study of phenotypic and genetic diversity in landrace collections is important for germplasm conservation and biodiversity protection. Here, we set up to study the germplasm collected in the region of Lazio in Central Italy with a focus on the distinctiveness among landraces and the attribution of membership to unnamed accessions. Our regional collection included 30 accessions belonging to six different locally recognized landraces and 21 unnamed accessions. All accessions were gathered in Lazio and belonged to the collection held at the Regional Agency for the Development and Innovation of Agriculture in Lazio (ARSIAL, in the application of the Regional Act n. 15/2000, funded by Lazio Rural Development Plan 2014 – 2020 Agro-environmental Measure, Action 10.2.1) and at the University of Tuscia. We included 13 control genotypes as references. The collection showed wide phenotypic variability for several traits, such as fruit weight (range 14-277 g), locule number (2-12), shape index (0.54-2.65), yield (0.24-3.08 kg/plant), and soluble solids (3.4-7.5 °B). A few landraces showed uncommon phenotypes, such as potato leaf, colorless fruit epidermis, or delayed ripening. Multivariate analysis of 25 cardinal phenotypic variables grouped the named varieties and allowed to assign of some of the unnamed to recognized groups. A case study for distinctiveness is presented for the flattened-ribbed types that presented overlapping distribution according to the phenotypic data. Molecular markers retrieved by previous studies revealed differences compared to the phenotyping clustering, indicating that the named varieties “Scatolone di Bolsena” and “Pantano Romanesco” belong to the Marmande group, together with the reference landrace from Tuscany “Costoluto Fiorentino”. Differently, the landrace “Spagnoletta di Formia e Gaeta” was clearly distinct from the former at the molecular level. Therefore, a genotypic analysis of the analyzed collection appears needed to better define the molecular distinctiveness among the flattened-ribbed accessions, as well as to properly attribute the membership group of the unnamed accessions.

Keywords: distinctiveness, flattened-ribbed fruits, regional landraces, tomato

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520 Pentosan Polysulfate Sodium: A Potential Treatment to Improve Bone and Joint Manifestations of Mucopolysaccharidosis I

Authors: Drago Bratkovic, Curtis Gravance, David Ketteridge, Ravi Krishnan, Michael Imperiale

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The mucopolysaccharidoses (MPSs) are a group of lysosomal storage diseases that have a common defect in the catabolism of glycosaminoglycans (GAGs). MPS I is the most common of the MPS diseases. Manifestations of MPS I include coarsening of facial features, corneal clouding, developmental delay, short stature, skeletal manifestations, hearing loss, cardiac valve disease, hepatosplenomegaly, and umbilical and inguinal hernias. Treatments for MPS I restore or activate the missing or deficient enzyme in the case of enzyme replacement therapy (ERT) and haematopoietic stem cell transplantation (HSCT). Pentosan polysulfate sodium (PPS) is a potential treatment to improve bone and joint manifestations of MPS I. The mechanisms of action of PPS that are relevant to the treatment of MPS I are the ability to: (i) Reduce systemic and accumulated GAG, (ii) Reduce inflammatory effects via the inhibition of NF-kB, resulting in the reduction in pro-inflammatory mediators. (iii) Reduce the expression of the pain mediator nerve growth factor in osteocytes from degenerating joints. (iv) Inhibit the cartilage degrading enzymes related to joint dysfunction in MPS I. PPS is being evaluated as an adjunctive therapy to ERT and/or HSCT in an open-label, single-centre, phase 2 study. Patients are ≥ 5 years of age with a diagnosis of MPS I and previously received HSCT and/or ERT. Three white, female, patients with MPS I-Hurler, ages 14, 15, and 19 years, and one, white male patient aged 15 years are enrolled. All were diagnosed at ≤2 years of age. All patients received HSCT ≤ 6 months after diagnosis. Two of the patients were treated with ERT prior to HSCT, and 1 patient received ERT commencing 3 months prior to HSCT. Two patients received 0.75mg/kg and 2 patients received 1.5mg/kg of PPS. PPS was well tolerated at doses of 0.75 and 1.5 mg/kg to 47 weeks of continuous dosing. Of the 19 adverse events (AEs), 2 were related to PPS. One AE was moderate (pre-syncope) and 1 was mild (injection site bruising), experienced in the same patient. All AEs were reported as mild or moderate. There have been no SAEs. One subject experienced a COVID-19 infection and PPS was interrupted. The MPS I signature GAG fragments, sulfated disaccharide and UA-HNAc S, tended to decrease in 3 patients from baseline through Week 25. Week 25 GAG data are pending for the 4th patient. Overall, most biomarkers (inflammatory, cartilage degeneration, and bone turnover) evaluated in the 3 patients with 25-week assessments have indicated either no change or a reduction in levels compared to baseline. In 3 patients, there was a trend toward improvement in the 2MWT from baseline to Week 48 with > 100% increase in 1 patient (01-201). In the 3 patients that had Week 48 assessments, patients and proxies reported improvement in PGIC, including “worthwhile difference” (n=1), or “made all the difference” (n=2).

Keywords: MPS I, pentosan polysulfate sodium, clinical study, 2MWT, QoL

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519 On Stochastic Models for Fine-Scale Rainfall Based on Doubly Stochastic Poisson Processes

Authors: Nadarajah I. Ramesh

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

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

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518 Genome-Wide Homozygosity Analysis of the Longevous Phenotype in the Amish Population

Authors: Sandra Smieszek, Jonathan Haines

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Introduction: Numerous research efforts have focused on searching for ‘longevity genes’. However, attempting to decipher the genetic component of the longevous phenotype have resulted in limited success and the mechanisms governing longevity remain to be explained. We conducted a genome-wide homozygosity analysis (GWHA) of the founder population of the Amish community in central Ohio. While genome-wide association studies using unrelated individuals have revealed many interesting longevity associated variants, these variants are typically of small effect and cannot explain the observed patterns of heritability for this complex trait. The Amish provide a large cohort of extended kinships allowing for in depth analysis via family-based approach excellent population due to its. Heritability of longevity increases with age with significant genetic contribution being seen in individuals living beyond 60 years of age. In our present analysis we show that the heritability of longevity is estimated to be increasing with age particularly on the paternal side. Methods: The present analysis integrated both phenotypic and genotypic data and led to the discovery of a series of variants, distinct for stratified populations across ages and distinct for paternal and maternal cohorts. Specifically 5437 subjects were analyzed and a subset of 893 successfully genotyped individuals was used to assess CHIP heritability. We have conducted the homozygosity analysis to examine if homozygosity is associated with increased risk of living beyond 90. We analyzed AMISH cohort genotyped for 614,957 SNPs. Results: We delineated 10 significant regions of homozygosity (ROH) specific for the age group of interest (>90). Of particular interest was ROH on chromosome 13, P < 0.0001. The lead SNPs rs7318486 and rs9645914 point to COL4A2 and our lead SNP. COL25A1 encodes one of the six subunits of type IV collagen, the C-terminal portion of the protein, known as canstatin, is an inhibitor of angiogenesis and tumor growth. COL4A2 mutations have been reported with a broader spectrum of cerebrovascular, renal, ophthalmological, cardiac, and muscular abnormalities. The second region of interest points to IRS2. Furthermore we built a classifier using the obtained SNPs from the significant ROH region with 0.945 AUC giving ability to discriminate between those living beyond to 90 years of age and beyond. Conclusion: In conclusion our results suggest that a history of longevity does indeed contribute to increasing the odds of individual longevity. Preliminary results are consistent with conjecture that heritability of longevity is substantial when we start looking at oldest fifth and smaller percentiles of survival specifically in males. We will validate all the candidate variants in independent cohorts of centenarians, to test whether they are robustly associated with human longevity. The identified regions of interest via ROH analysis could be of profound importance for the understanding of genetic underpinnings of longevity.

Keywords: regions of homozygosity, longevity, SNP, Amish

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517 Evaluation of Antioxidant Activity and Total Phenolic Content of Lens Esculenta Moench, Seeds

Authors: Vivek Kumar Gupta, Kripi Vohra, Monika Gupta

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Pulses have been a vital ingredient of the balanced human diet in India. Lentil (Lens culinaris Medikus or Lens esculenta Moench.) is a common legume known since biblical times. Lentil seeds, with or without hulls, are cooked as dhal and this has been the main dish for millennia in the South Asian region. Oxidative stress can damage lipids, proteins, enzymes, carbohydrates and DNA in cells and tissues, resulting in membrane damage, fragmentation or random cross linking of molecules like DNA, enzymes and structural proteins and even lead to cell death induced by DNA fragmentation and lipid peroxidation. These consequences of oxidative stress construct the molecular basis in the development of cancer, neurodegenerative disorders, cardiovascular diseases, diabetes and autoimmune. The aim of the present work is to assess the antioxidant potential of the peteroleum ether, acetone, methanol and water extract of the Lens esculenta seeds. In vitro antioxidant assessment of the extracts was carried out using 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, hydroxyl radical scavenging activity, reducing power assay. The quantitative estimation of total phenolic content, total flavonoid content in extracts and in plant material, total saponin content, total alkaloid content, crude fibre content, total volatile content, fat content and mucilage content in drug material was also carried out. Though all the extracts exhibited dose dependent reducing power activity the acetone extract was found to possess significant hydrogen donating ability in DPPH (45.83%-93.13%) and hydroxyl radical scavenging system (28.7%-46.41%) than the peteroleum ether, methanol and water extracts. Total phenolic content in the acetone and methanol extract was found to be 608 and 188 mg gallic acid equivalent of phenol/g of sample respectively. Total flavonoid content of acetone and methanol extract was found to be 128 and 30.6 mg quercetin equivalent/g of sample respectively. It is evident that acetone extract of Lentil seeds possess high levels of polyphenolics and flavonoids that could be utilized as antioxidants and neutraceuticals.

Keywords: antioxidant, flavanoids, Lens esculenta, polyphenols

Procedia PDF Downloads 466
516 Antimicrobial Resistance of Acinetobacter baumannii in Veterinary Settings: A One Health Perspective from Punjab, Pakistan

Authors: Minhas Alam, Muhammad Hidayat Rasool, Mohsin Khurshid, Bilal Aslam

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The genus Acinetobacter has emerged as a significant concern in hospital-acquired infections, particularly due to the versatility of Acinetobacter baumannii in causing nosocomial infections. The organism's remarkable metabolic adaptability allows it to thrive in various environments, including the environment, animals, and humans. However, the extent of antimicrobial resistance in Acinetobacter species from veterinary settings, especially in developing countries like Pakistan, remains unclear. This study aimed to isolate and characterize Acinetobacter spp. from veterinary settings in Punjab, Pakistan. A total of 2,230 specimens were collected, including 1,960 samples from veterinary settings (nasal and rectal swabs from dairy and beef cattle), 200 from the environment, and 70 from human clinical settings. Isolates were identified using routine microbiological procedures and confirmed by polymerase chain reaction (PCR). Antimicrobial susceptibility was determined by the disc diffusion method, and minimum inhibitory concentration (MIC) was measured by the micro broth dilution method. Molecular techniques, such as PCR and DNA sequencing, were used to screen for antimicrobial-resistant determinants. Genetic diversity was assessed using standard techniques. The results showed that the overall prevalence of A. baumannii in cattle was 6.63% (65/980). However, among cattle, a higher prevalence of A. baumannii was observed in dairy cattle, 7.38% (54/731), followed by beef cattle, 4.41% (11/249). Out of 65 A. baumannii isolates, the carbapenem resistance was found in 18 strains, i.e. 27.7%. The prevalence of A. baumannii in nasopharyngeal swabs was higher, i.e., 87.7% (57/65), as compared to rectal swabs, 12.3% (8/65). Class D β-lactamases genes blaOXA-23 and blaOXA-51 were present in all the CRAB from cattle. Among carbapenem-resistant isolates, 94.4% (17/18) were positive for class B β-lactamases gene blaIMP, whereas the blaNDM-1 gene was detected in only one isolate of A. baumannii. Among 70 clinical isolates of A. baumannii, 58/70 (82.9%) were positive for the blaOXA-23-like gene, and 87.1% (61/70) were CRAB isolates. Among all clinical isolates of A. baumannii, blaOXA-51-like gene was present. Hence, the co-existence of blaOXA-23 and blaOXA-51 was found in 82.85% of clinical isolates. From the environmental settings, a total of 18 A. baumannii isolates were recovered; among these, 38.88% (7/18) strains showed carbapenem resistance. All environmental isolates of A. baumannii harbored class D β-lactamases genes, i.e., blaOXA-51 and blaOXA-23 were detected in 38.9% (7/18) isolates. Hence, the co-existence of blaOXA-23 and blaOXA-51 was found in 38.88% of isolates. From environmental settings, 18 A. baumannii isolates were recovered, with 38.88% showing carbapenem resistance. All environmental isolates harbored blaOXA-51 and blaOXA-23 genes, with co-existence in 38.88% of isolates. MLST results showed ten different sequence types (ST) in clinical isolates, with ST 589 being the most common in carbapenem-resistant isolates. In veterinary isolates, ST2 was most common in CRAB isolates from cattle. Immediate control measures are needed to prevent the transmission of CRAB isolates among animals, the environment, and humans. Further studies are warranted to understand the mechanisms of antibiotic resistance spread and implement effective disease control programs.

Keywords: Acinetobacter baumannii, carbapenemases, drug resistance, MSLT

Procedia PDF Downloads 54
515 Examination of Porcine Gastric Biomechanics in the Antrum Region

Authors: Sif J. Friis, Mette Poulsen, Torben Strom Hansen, Peter Herskind, Jens V. Nygaard

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Gastric biomechanics governs a large range of scientific and engineering fields, from gastric health issues to interaction mechanisms between external devices and the tissue. Determination of mechanical properties of the stomach is, thus, crucial, both for understanding gastric pathologies as well as for the development of medical concepts and device designs. Although the field of gastric biomechanics is emerging, advances within medical devices interacting with the gastric tissue could greatly benefit from an increased understanding of tissue anisotropy and heterogeneity. Thus, in this study, uniaxial tensile tests of gastric tissue were executed in order to study biomechanical properties within the same individual as well as across individuals. With biomechanical tests in the strain domain, tissue from the antrum region of six porcine stomachs was tested using eight samples from each stomach (n = 48). The samples were cut so that they followed dominant fiber orientations. Accordingly, from each stomach, four samples were longitudinally oriented, and four samples were circumferentially oriented. A step-wise stress relaxation test with five incremental steps up to 25 % strain with 200 s rest periods for each step was performed, followed by a 25 % strain ramp test with three different strain rates. Theoretical analysis of the data provided stress-strain/time curves as well as 20 material parameters (e.g., stiffness coefficients, dissipative energy densities, and relaxation time coefficients) used for statistical comparisons between samples from the same stomach as well as in between stomachs. Results showed that, for the 20 material parameters, heterogeneity across individuals, when extracting samples from the same area, was in the same order of variation as the samples within the same stomach. For samples from the same stomach, the mean deviation percentage for all 20 parameters was 21 % and 18 % for longitudinal and circumferential orientations, compared to 25 % and 19 %, respectively, for samples across individuals. This observation was also supported by a nonparametric one-way ANOVA analysis, where results showed that the 20 material parameters from each of the six stomachs came from the same distribution with a level of statistical significance of P > 0.05. Direction-dependency was also examined, and it was found that the maximum stress for longitudinal samples was significantly higher than for circumferential samples. However, there were no significant differences in the 20 material parameters, with the exception of the equilibrium stiffness coefficient (P = 0.0039) and two other stiffness coefficients found from the relaxation tests (P = 0.0065, 0.0374). Nor did the stomach tissue show any significant differences between the three strain-rates used in the ramp test. Heterogeneity within the same region has not been examined earlier, yet, the importance of the sampling area has been demonstrated in this study. All material parameters found are essential to understand the passive mechanics of the stomach and may be used for mathematical and computational modeling. Additionally, an extension of the protocol used may be relevant for compiling a comparative study between the human stomach and the pig stomach.

Keywords: antrum region, gastric biomechanics, loading-unloading, stress relaxation, uniaxial tensile testing

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514 Advancing Equitable Healthcare for Trans and Gender-Diverse Students: A Community-Based Participatory Action Project

Authors: Al Huuskonen, Clio Lake, K. M. Naude, Polina Petlitsyna, Sorsha Henning, Julia Wimmers-Klick

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This project presents the outcomes of a community-based participatory action initiative aimed at advocating for equitable healthcare and human rights for trans, two-spirit, and gender-diverse individuals, building upon the University of British Columbia (UBC) Trans Coalition's ongoing efforts. Participatory Action Research (PAR) was chosen as the research method with the goal of improving trans rights on the UBC campus, particularly regarding equitable access to healthcare. PAR involves active community contribution throughout the research process, which in this case was done by way of liaising with student resource groups and advocacy leaders. The goals of this project were as follows: a) identify gaps in gender-affirming healthcare for UBC students by consulting the community and collaborating with UBC services, b) develop an information package outlining provincial and university-based health insurance for gender-affirming care (including hormone therapy and surgeries), FAQs, and resources for UBC's trans students, c) make this package available to UBC students and other national transgender advocacy organizations. The initiative successfully expanded the UBC AMS Student Health and Dental Plan to include gender-affirming procedural coverage, developed a care access guide for students, and advocated for improved health records inclusivity, mechanisms for trans students to report negative care experiences, and increased access to gender-affirming primary care through the on-campus health clinic. Collaboration with other universities' pride organizations and Trans Care BC yielded positive outcomes through broader coalition building and resource sharing. Ongoing efforts are underway to update provincial policies, particularly through expanding coverage under fair pharma care and addressing the compounding effects of the primary care crisis for trans individuals. The project's tangible results include improved trans rights on campus, especially in terms of healthcare access. Expanding healthcare coverage through student care benefits thousands of students, making the ability to undergo important affirming procedures more affordable. Providing students with information on extended coverage options and communication with their doctors further removes barriers to care and positively impacts student wellbeing. This initiative demonstrates the effectiveness of community-based participatory action in advancing equitable healthcare for trans and gender-diverse individuals and serves as a model for other institutions and organizations striving to promote inclusivity and advocate for marginalized populations' rights.

Keywords: equitable healthcare, trans and gender-diverse individuals, inclusivity, participatory action research project

Procedia PDF Downloads 80
513 Electrical Degradation of GaN-based p-channel HFETs Under Dynamic Electrical Stress

Authors: Xuerui Niu, Bolin Wang, Xinchuang Zhang, Xiaohua Ma, Bin Hou, Ling Yang

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The application of discrete GaN-based power switches requires the collaboration of silicon-based peripheral circuit structures. However, the packages and interconnection between the Si and GaN devices can introduce parasitic effects to the circuit, which has great impacts on GaN power transistors. GaN-based monolithic power integration technology is an emerging solution which can improve the stability of circuits and allow the GaN-based devices to achieve more functions. Complementary logic circuits consisting of GaN-based E-mode p-channel heterostructure field-effect transistors (p-HFETs) and E-mode n-channel HEMTs can be served as the gate drivers. E-mode p-HFETs with recessed gate have attracted increasing interest because of the low leakage current and large gate swing. However, they suffer from a poor interface between the gate dielectric and polarized nitride layers. The reliability of p-HFETs is analyzed and discussed in this work. In circuit applications, the inverter is always operated with dynamic gate voltage (VGS) rather than a constant VGS. Therefore, dynamic electrical stress has been simulated to resemble the operation conditions for E-mode p-HFETs. The dynamic electrical stress condition is as follows. VGS is a square waveform switching from -5 V to 0 V, VDS is fixed, and the source grounded. The frequency of the square waveform is 100kHz with the rising/falling time of 100 ns and duty ratio of 50%. The effective stress time is 1000s. A number of stress tests are carried out. The stress was briefly interrupted to measure the linear IDS-VGS, saturation IDS-VGS, As VGS switches from -5 V to 0 V and VDS = 0 V, devices are under negative-bias-instability (NBI) condition. Holes are trapped at the interface of oxide layer and GaN channel layer, which results in the reduction of VTH. The negative shift of VTH is serious at the first 10s and then changes slightly with the following stress time. However, different phenomenon is observed when VDS reduces to -5V. VTH shifts negatively during stress condition, and the variation in VTH increases with time, which is different from that when VDS is 0V. Two mechanisms exists in this condition. On the one hand, the electric field in the gate region is influenced by the drain voltage, so that the trapping behavior of holes in the gate region changes. The impact of the gate voltage is weakened. On the other hand, large drain voltage can induce the hot holes generation and lead to serious hot carrier stress (HCS) degradation with time. The poor-quality interface between the oxide layer and GaN channel layer at the gate region makes a major contribution to the high-density interface traps, which will greatly influence the reliability of devices. These results emphasize that the improved etching and pretreatment processes needs to be developed so that high-performance GaN complementary logics with enhanced stability can be achieved.

Keywords: GaN-based E-mode p-HFETs, dynamic electric stress, threshold voltage, monolithic power integration technology

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512 Properties of Sustainable Artificial Lightweight Aggregate

Authors: Wasan Ismail Khalil, Hisham Khalid Ahmed, Zainab Ali

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Structural Lightweight Aggregate Concrete (SLWAC) has been developed in recent years because it reduces the dead load, cost, thermal conductivity and coefficient of thermal expansion of the structure. So SLWAC has the advantage of being a relatively green building material. Lightweight Aggregate (LWA) is either occurs as natural material such as pumice, scoria, etc. or as artificial material produced from different raw materials such as expanded shale, clay, slate, etc. The use of SLWAC in Iraq is limited due to the lack in natural LWA. The existence of Iraqi clay deposit with different types and characteristics leads to the idea of producing artificial expanded clay aggregate. The main aim in this work is to present of the properties of artificial LWA produced in the laboratory. Available local bentonite clay which occurs in the Western region of Iraq was used as raw material to produce the LWA. Sodium silicate as liquid industrial waste material from glass plant was mixed with bentonite clay in mix proportion 1:1 by weight. The manufacturing method of the lightweight aggregate including, preparation and mixing of clay and sodium silicate, burning of the mixture in the furnace at the temperature between 750-800˚C for two hours, and finally gradually cooling process. The produced LWA was then crushed to small pieces then screened on standard sieve series and prepared with grading which conforms to the specifications of LWA. The maximum aggregate size used in this investigation is 10 mm. The chemical composition and the physical properties of the produced LWA are investigated. The results indicate that the specific gravity of the produced LWA is 1.5 with the density of 543kg/m3 and water absorption of 20.7% which is in conformity with the international standard of LWA. Many trail mixes were carried out in order to produce LWAC containing the artificial LWA produced in this research. The selected mix proportion is 1:1.5:2 (cement: sand: aggregate) by weight with water to cement ratio of 0.45. The experimental results show that LWAC has oven dry density of 1720 kg/m3, water absorption of 8.5%, the thermal conductivity of 0.723 W/m.K and compressive strength of 23 N/mm2. The SLWAC produced in this research can be used in the construction of different thermal insulated buildings and masonry units. It can be concluded that the SLWA produced in this study contributes to sustainable development by, using industrial waste materials, conserving energy, enhancing the thermal and structural efficiency of concrete.

Keywords: expanded clay, lightweight aggregate, structural lightweight aggregate concrete, sustainable

Procedia PDF Downloads 317
511 Hampering The 'Right to Know': Consequences of the Excessive Interpretation of the Notion of Exemption from the Right to Information

Authors: Tomasz Lewinski

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The right to know becomes gradually recognised as an increasing number of states adopts national legislations regarding access to state-held information. Laws differ from each other in the scope of the right to information (hereinafter: RTI). In all regimes of RTI, there are exceptions from the general notion of the right. States’ authorities too often use exceptions to justify refusals to requests for state-held information. This paper sets out how states hamper RTI basing on the notion of exception and by not providing an effective procedure that could redress unlawful denials. This paper bases on two selected examples of RTI incorporation into the national legal regime, United Kingdom, and South Africa. It succinctly outlines the international standard given in Article 19 of the International Covenant on Civil and Political Rights (hereinafter: ICCPR) and its influence on the RTI in selected countries. It shortly demonstrates as a background to further analysis the Human Rights Committee’s jurisprudence and standards articulated by successive Special Rapporteurs on freedom of opinion and expression. Subsequently, it presents a brief comparison of these standards with the regional standards, namely the African Charter on Human and Peoples' Rights and the European Convention on Human Rights. It critically discusses the regimes of exceptions in RTI legislations in respective national laws. It shows how excessive these regimes are, what implications they have for the transparency in general. Also, the objective is to divide exceptions enumerated in legislations of selected states in relation to exceptions provided in Article 19 of the ICCPR. Basing on the established division of exceptions by its natures, it compares both regimes of exceptions related to the principle of national security. That is to compare jurisprudence of domestic courts, and overview practices of states’ authorities applied to RTI requests. The paper evaluates remedies available in legislations, including contexts of the length and costs of the subsequent proceedings. This provides a general assessment of the given mechanisms and present potential risks of its ineffectiveness. The paper relies on examination of the national legislations, comments of the credible non-governmental organisations (e.g. The Public's Right to Know Principles on Freedom of Information Legislation by the Article 19, The Tshwane Principles on National Security and the Right to Information), academics and also the research of the relevant judgements delivered by domestic and international courts. Conclusion assesses whether selected countries’ legislations go in line with international law and trends, whether the jurisprudence of the regional courts provide appropriate benchmarks for national courts to address RTI issues effectively. Furthermore, it identifies the largest disadvantages of current legislations and to what outcomes it leads in domestic courts jurisprudences. In the end, it provides recommendations and policy arguments for states to improve transparency and support local organisations in their endeavours to establish more transparent states and societies.

Keywords: access to information, freedom of information, national security, right to know, transparency

Procedia PDF Downloads 199
510 Kinetic Studies on CO₂ Gasification of Low and High Ash Indian Coals in Context of Underground Coal Gasification

Authors: Geeta Kumari, Prabu Vairakannu

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Underground coal gasification (UCG) technology is an efficient and an economic in-situ clean coal technology, which converts unmineable coals into calorific valuable gases. This technology avoids ash disposal, coal mining, and storage problems. CO₂ gas can be a potential gasifying medium for UCG. CO₂ is a greenhouse gas and, the liberation of this gas to the atmosphere from thermal power plant industries leads to global warming. Hence, the capture and reutilization of CO₂ gas are crucial for clean energy production. However, the reactivity of high ash Indian coals with CO₂ needs to be assessed. In the present study, two varieties of Indian coals (low ash and high ash) are used for thermogravimetric analyses (TGA). Two low ash north east Indian coals (LAC) and a typical high ash Indian coal (HAC) are procured from the coal mines of India. Low ash coal with 9% ash (LAC-1) and 4% ash (LAC-2) and high ash coal (HAC) with 42% ash are used for the study. TGA studies are carried out to evaluate the activation energy for pyrolysis and gasification of coal under N₂ and CO₂ atmosphere. Coats and Redfern method is used to estimate the activation energy of coal under different temperature regimes. Volumetric model is assumed for the estimation of the activation energy. The activation energy estimated under different temperature range. The inherent properties of coals play a major role in their reactivity. The results show that the activation energy decreases with the decrease in the inherent percentage of coal ash due to the ash layer hindrance. A reverse trend was observed with volatile matter. High volatile matter of coal leads to the estimation of low activation energy. It was observed that the activation energy under CO₂ atmosphere at 400-600°C is less as compared to N₂ inert atmosphere. At this temperature range, it is estimated that 15-23% reduction in the activation energy under CO₂ atmosphere. This shows the reactivity of CO₂ gas with higher hydrocarbons of the coal volatile matters. The reactivity of CO₂ with the volatile matter of coal might occur through dry reforming reaction in which CO₂ reacts with higher hydrocarbon, aromatics of the tar content. The observed trend of Ea in the temperature range of 150-200˚C and 400-600˚C is HAC > LAC-1 >LAC-2 in both N₂ and CO₂ atmosphere. At the temperature range of 850-1000˚C, higher activation energy is estimated when compared to those values in the temperature range of 400-600°C. Above 800°C, char gasification through Boudouard reaction progressed under CO₂ atmosphere. It was observed that 8-20 kJ/mol of activation energy is increased during char gasification above 800°C compared to volatile matter pyrolysis between the temperature ranges of 400-600°C. The overall activation energy of the coals in the temperature range of 30-1000˚C is higher in N₂ atmosphere than CO₂ atmosphere. It can be concluded that higher hydrocarbons such as tar effectively undergoes cracking and reforming reactions in presence of CO₂. Thus, CO₂ gas is beneficial for the production of high calorific value syngas using high ash Indian coals.

Keywords: clean coal technology, CO₂ gasification, activation energy, underground coal gasification

Procedia PDF Downloads 159
509 Enhancing of Antibacterial Activity of Essential Oil by Rotating Magnetic Field

Authors: Tomasz Borowski, Dawid Sołoducha, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

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Essential oils (EOs) are fragrant volatile oils obtained from plants. These are used for cooking (for flavor and aroma), cleaning, beauty (e.g., rosemary essential oil is used to promote hair growth), health (e.g. thyme essential oil cures arthritis, normalizes blood pressure, reduces stress on the heart, cures chest infection and cough) and in the food industry as preservatives and antioxidants. Rosemary and thyme essential oils are considered the most eminent herbs based on their history and medicinal properties. They possess a wide range of activity against different types of bacteria and fungi compared with the other oils in both in vitro and in vivo studies. However, traditional uses of EOs are limited due to rosemary and thyme oils in high concentrations can be toxic. In light of the accessible data, the following hypothesis was put forward: Low frequency rotating magnetic field (RMF) increases the antimicrobial potential of EOs. The aim of this work was to investigate the antimicrobial activity of commercial Salvia Rosmarinus L. and Thymus vulgaris L. essential oil from Polish company Avicenna-Oil under Rotating Magnetic Field (RMF) at f = 25 Hz. The self-constructed reactor (MAP) was applied for this study. The chemical composition of oils was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Minimum inhibitory concentrations (MIC) against E. coli were determined for the essential oils. Tested oils in very small concentrations were prepared (from 1 to 3 drops of essential oils per 3 mL working suspensions). From the results of disc diffusion assay and MIC tests, it can be concluded that thyme oil had the highest antibacterial activity against E. coli. Moreover, the study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of tested oils. The extended radiation exposure to RMF at the frequency f= 25 Hz beyond 160 minutes resulted in a significant increase in antibacterial potential against E. coli. Bacteria were killed within 40 minutes in thyme oil in lower tested concentration (1 drop of essential oils per 3 mL working suspension). Rapid decrease (>3 log) of bacteria number was observed with rosemary oil within 100 minutes (in concentration 3 drops of essential oils per 3 mL working suspension). Thus, a method for improving the antimicrobial performance of essential oil in low concentrations was developed. However, it still remains to be investigated how bacteria get killed by the EOs treated by an electromagnetic field. The possible mechanisms relies on alteration in the permeability of ionic channels in ionic channels in the bacterial cell walls that transport in the cells was proposed. For further studies, it is proposed to examine other types of essential oils and other antibiotic-resistant bacteria (ARB), which are causing a serious concern throughout the world.

Keywords: rotating magnetic field, rosemary, thyme, essential oils, Escherichia coli

Procedia PDF Downloads 148
508 Re-Presenting the Egyptian Informal Urbanism in Films between 1994 and 2014

Authors: R. Mofeed, N. Elgendy

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Cinema constructs mind-spaces that reflect inherent human thoughts and emotions. As a representational art, Cinema would introduce comprehensive images of life phenomena in different ways. The term “represent” suggests verity of meanings; bring into presence, replace or typify. In that sense, Cinema may present a phenomenon through direct embodiment, or introduce a substitute image that replaces the original phenomena, or typify it by relating the produced image to a more general category through a process of abstraction. This research is interested in questioning the type of images that Egyptian Cinema introduces to informal urbanism and how these images were conditioned and reshaped in the last twenty years. The informalities/slums phenomenon first appeared in Egypt and, particularly, Cairo in the early sixties, however, this phenomenon was completely ignored by the state and society until the eighties, and furthermore, its evident representation in Cinema was by the mid-nineties. The Informal City represents the illegal housing developments, and it is a fast growing form of urbanization in Cairo. Yet, this expanding phenomenon is still depicted as the minority, exceptional and marginal through the Cinematic lenses. This paper aims at tracing the forms of representations of the urban informalities in the Egyptian Cinema between 1994 and 2014, and how did that affect the popular mind and its perception of these areas. The paper runs two main lines of inquiry; the first traces the phenomena through a chronological and geographical mapping of the informal urbanism has been portrayed in films. This analysis is based on an academic research work at Cairo University in Fall 2014. The visual tracing through maps and timelines allowed a reading of the phases of ignorance, presence, typifying and repetition in the representation of this huge sector of the city through more than 50 films that has been investigated. The analysis clearly revealed the “portrayed image” of informality by the Cinema through the examined period. However, the second part of the paper explores the “perceived image”. A designed questionnaire is applied to highlight the main features of that image that is perceived by both inhabitants of informalities and other Cairenes based on watching selected films. The questionnaire covers the different images of informalities proposed in the Cinema whether in a comic or a melodramatic background and highlight the descriptive terms used, to see which of them resonate with the mass perceptions and affected their mental images. The two images; “portrayed” and “perceived” are then to be encountered to reflect on issues of repetitions, stereotyping and reality. The formulated stereotype of informal urbanism is finally outlined and justified in relation to both production consumption mechanisms of films and the State official vision of informalities.

Keywords: cinema, informal urbanism, popular mind, representation

Procedia PDF Downloads 287
507 Identification and Characterization of Antimicrobial Peptides Isolated from Entophytic Bacteria and Their Activity against Multidrug-Resistance Gram-Negative Bacteria in South Korea

Authors: Maryam Beiranvand

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Multi-drug resistance in various microorganisms has increased globally in many healthcare facilities. Less effective antimicrobial activity of drug therapies for infection control becomes trouble. Since 1980, no new type of antimicrobial drug has been identified, even though combinations of antibiotic drugs have been discovered almost every decade. Between 1981 and 2006, over 70% of novel pharmaceuticals and chemical agents came from natural sources. Microorganisms have yielded almost 22,000 natural compounds. The identification of antimicrobial components from endophytes bacteria could help overcome the threat posed by multi-drug resistant strains. The project aims to analyze and identify antimicrobial peptides isolated from entophytic bacteria and their activity against multidrug-resistant Gram-negative bacteria in South Korea. Endophytic Paenibacillus polymyxa. 4G3 isolated from the plant, Gynura procumbery exhibited considerable antimicrobial activity against Methicillin-resistant Staphylococcus aureus, and Escherichia coli. The Rapid Annotations using Subsystems Technology showed that the total size of the draft genome was 5,739,603bp, containing 5178 genes with 45.8% G+C content. Genome annotation using antiSMASH version 6.0.0 was performed, which predicted the most common types of non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS). In this study, diethyl aminoethyl cellulose (DEAEC) resin was used as the first step in purifying for unknown peptides, and then the target protein was identified using hydrophilic and hydrophobic solutions, optimal pH, and step-by-step tests for antimicrobial activity. This crude was subjected to C18 chromatography and elution with 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% methanol, respectively. Only the fraction eluted with 20% -60% methanol demonstrated good antimicrobial activity against MDR E. coli. The concentration of the active fragment was measured by the Brad-ford test, and Protein A280 - Thermo Fisher Scientific at the end by examining the SDS PAGE Resolving Gel, 10% Acrylamide and purity were confirmed. Our study showed that, based on the combined results of the analysis and purification. P polymyxa. 4G3 has a high potential exists for producing novel functions of polymyxin E and bacitracin against bacterial pathogens.

Keywords: endophytic bacteria, antimicrobial activity, antimicrobial peptide, whole genome sequencing analysis, multi -drug resistance gram negative bacteria

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506 Screening Tools and Its Accuracy for Common Soccer Injuries: A Systematic Review

Authors: R. Christopher, C. Brandt, N. Damons

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Background: The sequence of prevention model states that by constant assessment of injury, injury mechanisms and risk factors are identified, highlighting that collecting and recording of data is a core approach for preventing injuries. Several screening tools are available for use in the clinical setting. These screening techniques only recently received research attention, hence there is a dearth of inconsistent and controversial data regarding their applicability, validity, and reliability. Several systematic reviews related to common soccer injuries have been conducted; however, none of them addressed the screening tools for common soccer injuries. Objectives: The purpose of this study was to conduct a review of screening tools and their accuracy for common injuries in soccer. Methods: A systematic scoping review was performed based on the Joanna Briggs Institute procedure for conducting systematic reviews. Databases such as SPORT Discus, Cinahl, Medline, Science Direct, PubMed, and grey literature were used to access suitable studies. Some of the key search terms included: injury screening, screening, screening tool accuracy, injury prevalence, injury prediction, accuracy, validity, specificity, reliability, sensitivity. All types of English studies dating back to the year 2000 were included. Two blind independent reviewers selected and appraised articles on a 9-point scale for inclusion as well as for the risk of bias with the ACROBAT-NRSI tool. Data were extracted and summarized in tables. Plot data analysis was done, and sensitivity and specificity were analyzed with their respective 95% confidence intervals. I² statistic was used to determine the proportion of variation across studies. Results: The initial search yielded 95 studies, of which 21 were duplicates, and 54 excluded. A total of 10 observational studies were included for the analysis: 3 studies were analysed quantitatively while the remaining 7 were analysed qualitatively. Seven studies were graded low and three studies high risk of bias. Only high methodological studies (score > 9) were included for analysis. The pooled studies investigated tools such as the Functional Movement Screening (FMS™), the Landing Error Scoring System (LESS), the Tuck Jump Assessment, the Soccer Injury Movement Screening (SIMS), and the conventional hamstrings to quadriceps ratio. The accuracy of screening tools was of high reliability, sensitivity and specificity (calculated as ICC 0.68, 95% CI: 52-0.84; and 0.64, 95% CI: 0.61-0.66 respectively; I² = 13.2%, P=0.316). Conclusion: Based on the pooled results from the included studies, the FMS™ has a good inter-rater and intra-rater reliability. FMS™ is a screening tool capable of screening for common soccer injuries, and individual FMS™ scores are a better determinant of performance in comparison with the overall FMS™ score. Although meta-analysis could not be done for all the included screening tools, qualitative analysis also indicated good sensitivity and specificity of the individual tools. Higher levels of evidence are, however, needed for implication in evidence-based practice.

Keywords: accuracy, screening tools, sensitivity, soccer injuries, specificity

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505 An Assessment of Nodulation and Nitrogen Fixation of Lessertia Frutescens Plants Inoculated with Rhizobial Isolates from the Cape Fynbos

Authors: Mokgadi Miranda Hlongwane, Ntebogeng Sharon Mokgalaka, Felix Dapare Dakora

Abstract:

Lessertia (L.) frutescens (syn. Sutherlandia frutescens) is a leguminous medicinal plant indigenous to South Africa. Traditionally, L. frutescens has been used to treat cancer, diabetes, epilepsy, fever, HIV, stomach problems, wounds and other ailments. This legume is endemic to the Cape fynbos, with large populations occurring wild and cultivated in the Cape Florist Region. Its widespread distribution in the Western Cape, Northern Cape, Eastern Cape and Kwazulu-Natal is linked to its increased use as a phytomedicine in the treatment of various diseases by traditional healers. The frequent harvesting of field plants for use as a medicine has made it necessary to undertake studies towards the conservation of Lessertia frutescens. As a legume, this species can form root nodules and fix atmospheric N₂ when in symbiosis with soil bacteria called rhizobia. So far, however, few studies (if any) have been done on the efficacy and diversity of native bacterial symbionts nodulating L. frutescens in South Africa. The aim of this project was to isolate and characterize L. frutescens-nodulating bacteria from five different locations in the Western Cape Province. This was done by trapping soil rhizobia using rhizosphere soil suspension to inoculate L. frutescens seedlings growing in sterilized sand and receiving sterile N-free Hoagland nutrient solution under glasshouse conditions. At 60 days after planting, root nodules were harvested from L. frutescens plants, surface-sterilized, macerated, and streaked on yeast mannitol agar (YMA) plates and incubated at 28 ˚C for observation of bacterial growth. The majority of isolates were slow-growers that took 6-14 days to appear on YMA plates. However, seven isolates were fast-growers, taking 2-4 days to appear on YMA plates. Single-colony cultures of the isolates were assessed for their ability to nodulate L. frutescens as a homologous host under glasshouse conditions. Of the 92 bacterial isolates tested, 63 elicited nodule formation on L. frutescens. Symbiotic effectiveness varied markedly between and among test isolates. There were also significant (p≤0.005) differences in nodulation, shoot biomass, photosynthetic rates, leaf transpiration and stomatal conductance of L. frutescens plants inoculated with the test isolates, which is an indication of their functional diversity.

Keywords: lessertia frutescens, nodulating, rhizobia, symbiotic effectiveness

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504 Functional Analysis of Variants Implicated in Hearing Loss in a Cohort from Argentina: From Molecular Diagnosis to Pre-Clinical Research

Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

Abstract:

Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

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503 Carbon Capture and Storage by Continuous Production of CO₂ Hydrates Using a Network Mixing Technology

Authors: João Costa, Francisco Albuquerque, Ricardo J. Santos, Madalena M. Dias, José Carlos B. Lopes, Marcelo Costa

Abstract:

Nowadays, it is well recognized that carbon dioxide emissions, together with other greenhouse gases, are responsible for the dramatic climate changes that have been occurring over the past decades. Gas hydrates are currently seen as a promising and disruptive set of materials that can be used as a basis for developing new technologies for CO₂ capture and storage. Its potential as a clean and safe pathway for CCS is tremendous since it requires only water and gas to be mixed under favorable temperatures and mild high pressures. However, the hydrates formation process is highly exothermic; it releases about 2 MJ per kilogram of CO₂, and it only occurs in a narrow window of operational temperatures (0 - 10 °C) and pressures (15 to 40 bar). Efficient continuous hydrate production at a specific temperature range necessitates high heat transfer rates in mixing processes. Past technologies often struggled to meet this requirement, resulting in low productivity or extended mixing/contact times due to inadequate heat transfer rates, which consistently posed a limitation. Consequently, there is a need for more effective continuous hydrate production technologies in industrial applications. In this work, a network mixing continuous production technology has been shown to be viable for producing CO₂ hydrates. The structured mixer used throughout this work consists of a network of unit cells comprising mixing chambers interconnected by transport channels. These mixing features result in enhanced heat and mass transfer rates and high interfacial surface area. The mixer capacity emerges from the fact that, under proper hydrodynamic conditions, the flow inside the mixing chambers becomes fully chaotic and self-sustained oscillatory flow, inducing intense local laminar mixing. The device presents specific heat transfer rates ranging from 107 to 108 W⋅m⁻³⋅K⁻¹. A laboratory scale pilot installation was built using a device capable of continuously capturing 1 kg⋅h⁻¹ of CO₂, in an aqueous slurry of up to 20% in mass. The strong mixing intensity has proven to be sufficient to enhance dissolution and initiate hydrate crystallization without the need for external seeding mechanisms and to achieve, at the device outlet, conversions of 99% in CO₂. CO₂ dissolution experiments revealed that the overall liquid mass transfer coefficient is orders of magnitude larger than in similar devices with the same purpose, ranging from 1 000 to 12 000 h⁻¹. The present technology has shown itself to be capable of continuously producing CO₂ hydrates. Furthermore, the modular characteristics of the technology, where scalability is straightforward, underline the potential development of a modular hydrate-based CO₂ capture process for large-scale applications.

Keywords: network, mixing, hydrates, continuous process, carbon dioxide

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502 Soil Quality Response to Long-Term Intensive Resources Management and Soil Texture

Authors: Dalia Feiziene, Virginijus Feiza, Agne Putramentaite, Jonas Volungevicius, Kristina Amaleviciute, Sarunas Antanaitis

Abstract:

The investigations on soil conservation are one of the most important topics in modern agronomy. Soil management practices have great influence on soil physico-chemical quality and GHG emission. Research objective: To reveal the sensitivity and vitality of soils with different texture to long-term antropogenisation on Cambisol in Central Lithuania and to compare them with not antropogenised soil resources. Methods: Two long-term field experiments (loam on loam; sandy loam on loam) with different management intensity were estimated. Disturbed and undisturbed soil samples were collected from 5-10, 15-20 and 30-35 cm depths. Soil available P and K contents were determined by ammonium lactate extraction, total N by the dry combustion method, SOC content by Tyurin titrimetric (classical) method, texture by pipette method. In undisturbed core samples soil pore volume distribution, plant available water (PAW) content were determined. A closed chamber method was applied to quantify soil respiration (SR). Results: Long-term resources management changed soil quality. In soil with loam texture, within 0-10, 10-20 and 30-35 cm soil layers, significantly higher PAW, SOC and mesoporosity (MsP) were under no-tillage (NT) than under conventional tillage (CT). However, total porosity (TP) under NT was significantly higher only in 0-10 cm layer. MsP acted as dominant factor for N, P and K accumulation in adequate layers. P content in all soil layers was higher under NT than in CT. N and K contents were significantly higher than under CT only in 0-10 cm layer. In soil with sandy loam texture, significant increase in SOC, PAW, MsP, N, P and K under NT was only in 0-10 cm layer. TP under NT was significantly lower in all layers. PAW acted as strong dominant factor for N, P, K accumulation. The higher PAW the higher NPK contents were determined. NT did not secure chemical quality within deeper layers than CT. Long-term application of mineral fertilisers significantly increased SOC and soil NPK contents primarily in top-soil. Enlarged fertilization determined the significantly higher leaching of nutrients to deeper soil layers (CT) and increased hazards of top-soil pollution. Straw returning significantly increased SOC and NPK accumulation in top-soil. The SR on sandy loam was significantly higher than on loam. At dry weather conditions, on loam SR was higher in NT than in CT, on sandy loam SR was higher in CT than in NT. NPK fertilizers promoted significantly higher SR in both dry and wet year, but suppressed SR on sandy loam during usual year. Not antropogenised soil had similar SOC and NPK distribution within 0-35 cm layer and depended on genesis of soil profile horizons.

Keywords: fertilizers, long-term experiments, soil texture, soil tillage, straw

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501 A Biophysical Study of the Dynamic Properties of Glucagon Granules in α Cells by Imaging-Derived Mean Square Displacement and Single Particle Tracking Approaches

Authors: Samuele Ghignoli, Valentina de Lorenzi, Gianmarco Ferri, Stefano Luin, Francesco Cardarelli

Abstract:

Insulin and glucagon are the two essential hormones for maintaining proper blood glucose homeostasis, which is disrupted in Diabetes. A constantly growing research interest has been focused on the study of the subcellular structures involved in hormone secretion, namely insulin- and glucagon-containing granules, and on the mechanisms regulating their behaviour. Yet, while several successful attempts were reported describing the dynamic properties of insulin granules, little is known about their counterparts in α cells, the glucagon-containing granules. To fill this gap, we used αTC1 clone 9 cells as a model of α cells and ZIGIR as a fluorescent Zinc chelator for granule labelling. We started by using spatiotemporal fluorescence correlation spectroscopy in the form of imaging-derived mean square displacement (iMSD) analysis. This afforded quantitative information on the average dynamical and structural properties of glucagon granules having insulin granules as a benchmark. Interestingly, the iMSD sensitivity to average granule size allowed us to confirm that glucagon granules are smaller than insulin ones (~1.4 folds, further validated by STORM imaging). To investigate possible heterogeneities in granule dynamic properties, we moved from correlation spectroscopy to single particle tracking (SPT). We developed a MATLAB script to localize and track single granules with high spatial resolution. This enabled us to classify the glucagon granules, based on their dynamic properties, as ‘blocked’ (i.e., trajectories corresponding to immobile granules), ‘confined/diffusive’ (i.e., trajectories corresponding to slowly moving granules in a defined region of the cell), or ‘drifted’ (i.e., trajectories corresponding to fast-moving granules). In cell-culturing control conditions, results show this average distribution: 32.9 ± 9.3% blocked, 59.6 ± 9.3% conf/diff, and 7.4 ± 3.2% drifted. This benchmarking provided us with a foundation for investigating selected experimental conditions of interest, such as the glucagon-granule relationship with the cytoskeleton. For instance, if Nocodazole (10 μM) is used for microtubule depolymerization, the percentage of drifted motion collapses to 3.5 ± 1.7% while immobile granules increase to 56.0 ± 10.7% (remaining 40.4 ± 10.2% of conf/diff). This result confirms the clear link between glucagon-granule motion and cytoskeleton structures, a first step towards understanding the intracellular behaviour of this subcellular compartment. The information collected might now serve to support future investigations on glucagon granules in physiology and disease. Acknowledgment: This work has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 866127, project CAPTUR3D).

Keywords: glucagon granules, single particle tracking, correlation spectroscopy, ZIGIR

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