Search results for: engineered physiology

Authors: Mariam Dianat Sabet Gilani, Lars M. Blank, Birgitta E. Ebert

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Plant-derived lupane-type, pentacyclic triterpenoids are biologically active compounds that are highly interesting for applications in medical, pharmaceutical, and cosmetic industries. Due to the low abundance of these valuable compounds in their natural sources, and the environmentally harmful downstream process, alternative production methods, such as microbial cell factories, are investigated. Engineered Saccharomyces cerevisiae strains, harboring the heterologous genes for betulinic acid synthesis, can produce up to 2 g L-1 triterpenoids, showing high potential for large-scale production of triterpenoids. One limitation of the microbial synthesis is the intracellular product accumulation. It not only makes cell disruption a necessary step in the downstream processing but also limits productivity and product yield per cell. To overcome these restrictions, the aim of this study is to develop an in-situ extraction method, which extracts triterpenoids into a second organic phase. Such a continuous or sequential product removal from the biomass keeps the cells in an active state and enables extended production time or biomass recycling. After screening of twelve different solvents, selected based on product solubility, biocompatibility, as well as environmental and health impact, isopropyl myristate (IPM) was chosen as a suitable solvent for in-situ product removal from S. cerevisiae. Impedance-based single-cell analysis and off-gas measurement of carbon dioxide emission showed that cell viability and physiology were not affected by the presence of IPM. Initial experiments demonstrated that after the addition of 20 vol % IPM to cultures in the stationary phase, 40 % of the total produced triterpenoids were extracted from the cells into the organic phase. In future experiments, the application of IPM in a repeated batch process will be tested, where IPM is added at the end of each batch run to remove triterpenoids from the cells, allowing the same biocatalysts to be used in several sequential batch steps. Due to its high biocompatibility, the amount of IPM added to the culture can also be increased to more than 20 vol % to extract more than 40 % triterpenoids in the organic phase, allowing the cells to produce more triterpenoids. This highlights the potential for the development of a continuous large-scale process, which allows biocatalysts to produce intracellular products continuously without the necessity of cell disruption and without limitation of the cell capacity.

Keywords: betulinic acid, biocompatible solvent, in-situ extraction, isopropyl myristate, process development, secondary metabolites, triterpenoids, yeast

Procedia PDF Downloads 124

Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

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The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

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Authors: David Holton, Michelle Dickinson, Giovanni Carta

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The principle of geological disposal is to isolate higher-activity radioactive wastes deep inside a suitable rock formation to ensure that no harmful quantities of radioactivity reach the surface environment. To achieve this, wastes will be placed in an engineered underground containment facility – the geological disposal facility (GDF) – which will be designed so that natural and man-made barriers work together to minimise the escape of radioactivity. Internationally, various multi-barrier concepts have been developed for the disposal of higher-activity radioactive wastes. High-heat-generating wastes (HLW, spent fuel and Pu) provide a number of different technical challenges to those associated with the disposal of low-heat-generating waste. Thermal management of the disposal system must be taken into consideration in GDF design; temperature constraints might apply to the wasteform, container, buffer and host rock. Of these, the temperature limit placed on the buffer component of the engineered barrier system (EBS) can be the most constraining factor. The heat must therefore be managed such that the properties of the buffer are not compromised to the extent that it cannot deliver the required level of safety. The maximum temperature of a buffer surrounding a container at the centre of a fixed array of heat-generating sources, arises due to heat diffusing from neighbouring heat-generating wastes, incrementally contributing to the temperature of the EBS. A range of strategies can be employed for managing heat in a GDF, including the spatial arrangements or patterns of those containers; different geometrical configurations can influence the overall thermal density in a disposal facility (or area within a facility) and therefore the maximum buffer temperature. A semi-analytical thermal dimensioning tool and methodology have been applied at a generic stage to explore a range of strategies to manage the disposal of high-heat-generating waste. A number of examples, including different geometrical layouts and chequer-boarding, have been illustrated to demonstrate how these tools can be used to consider safety margins and inform strategic disposal options when faced with uncertainty, at a generic stage of the development of a GDF.

Keywords: buffer, geological disposal facility, high-heat-generating waste, spent fuel

Procedia PDF Downloads 257

Authors: Hassan Mahmoudzadeh

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The bacterium of Agrobacterium vitis causes crown and root gall disease, an important disease of grapevine, Vitis vinifera L. Also, Phylloxera is one of the most important pests in viticulture. Grapevine rootstocks were developed to provide increased resistance to soil-borne pests and diseases, but rootstock effects on some traits remain unclear. The interaction between rootstock, scion and environment can induce different responses to the grapevine physiology. 'Sultsna' (Vitis vinifera L.) is one of the most valuable raisin grape cultivars in Iran. Thus, the aim of this study was to determine the rootstock effect on the growth characteristics and yield components and quality of 'Sultana' grapevine grown in the Urmia viticulture region. The experimental design was completely randomized blocks, with four treatments, four replicates and 10 vines per plot. The results show that all variables evaluated were significantly affected by the rootstock. The Sultana/110R and Sultana/Nazmieh were among other combinations influenced by the year and had a higher significant yield/vine (13.25 and 12.14, respectively). Indeed, they were higher than that of Sultana/5BB (10.56 kg/vine) and Sultana/Spota (10.25 kg/vine). The number of clusters per burst bud and per vine and the weight of clusters were affected by the rootstock as well. Pruning weight/vine, yield/pruning weight, leaf area/vine and leaf area index are variables related to the physiology of grapevine, which was also affected by the rootstocks. In general, rootstocks had adapted well to the environment where the experiment was carried out, giving vigor and high yield to Sultana grapevine, which means that they may be used by grape growers in this region. In sum, the study found the best rootstocks for 'Sultana' to be Nazmieh and 110R in terms of root and shoot growth. However, the choice of the right rootstock depends on various aspects, such as those related to soil characteristics, climate conditions, grape varieties, and even clones, and production purposes.

Keywords: grafting, vineyards, grapevine, succeptability

Procedia PDF Downloads 88

Authors: Babak Forouraghi

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A genetic circuit is a collection of interacting genes and proteins that enable individual cells to implement and perform vital biological functions such as cell division, growth, death, and signaling. In cell engineering, synthetic gene circuits are engineered networks of genes specifically designed to implement functionalities that are not evolved by nature. These engineered networks enable scientists to tackle complex problems such as engineering cells to produce therapeutics within the patient's body, altering T cells to target cancer-related antigens for treatment, improving antibody production using engineered cells, tissue engineering, and production of genetically modified plants and livestock. Construction of computational models to realize genetic circuits is an especially challenging task since it requires the discovery of flow of genetic information in complex biological systems. Building synthetic biological models is also a time-consuming process with relatively low prediction accuracy for highly complex genetic circuits. The primary goal of this study was to investigate the utility of a pre-trained bidirectional encoder transformer that can accurately predict gene expressions in genetic circuit designs. The main reason behind using transformers is their innate ability (attention mechanism) to take account of the semantic context present in long DNA chains that are heavily dependent on spatial representation of their constituent genes. Previous approaches to gene circuit design, such as CNN and RNN architectures, are unable to capture semantic dependencies in long contexts as required in most real-world applications of synthetic biology. For instance, RNN models (LSTM, GRU), although able to learn long-term dependencies, greatly suffer from vanishing gradient and low-efficiency problem when they sequentially process past states and compresses contextual information into a bottleneck with long input sequences. In other words, these architectures are not equipped with the necessary attention mechanisms to follow a long chain of genes with thousands of tokens. To address the above-mentioned limitations of previous approaches, a transformer model was built in this work as a variation to the existing DNA Bidirectional Encoder Representations from Transformers (DNABERT) model. It is shown that the proposed transformer is capable of capturing contextual information from long input sequences with attention mechanism. In a previous work on genetic circuit design, the traditional approaches to classification and regression, such as Random Forrest, Support Vector Machine, and Artificial Neural Networks, were able to achieve reasonably high R2 accuracy levels of 0.95 to 0.97. However, the transformer model utilized in this work with its attention-based mechanism, was able to achieve a perfect accuracy level of 100%. Further, it is demonstrated that the efficiency of the transformer-based gene expression classifier is not dependent on presence of large amounts of training examples, which may be difficult to compile in many real-world gene circuit designs.

Keywords: transformers, generative ai, gene expression design, classification

Procedia PDF Downloads 38

Authors: Babak Forouraghi

Abstract:

A genetic circuit is a collection of interacting genes and proteins that enable individual cells to implement and perform vital biological functions such as cell division, growth, death, and signaling. In cell engineering, synthetic gene circuits are engineered networks of genes specifically designed to implement functionalities that are not evolved by nature. These engineered networks enable scientists to tackle complex problems such as engineering cells to produce therapeutics within the patient's body, altering T cells to target cancer-related antigens for treatment, improving antibody production using engineered cells, tissue engineering, and production of genetically modified plants and livestock. Construction of computational models to realize genetic circuits is an especially challenging task since it requires the discovery of the flow of genetic information in complex biological systems. Building synthetic biological models is also a time-consuming process with relatively low prediction accuracy for highly complex genetic circuits. The primary goal of this study was to investigate the utility of a pre-trained bidirectional encoder transformer that can accurately predict gene expressions in genetic circuit designs. The main reason behind using transformers is their innate ability (attention mechanism) to take account of the semantic context present in long DNA chains that are heavily dependent on the spatial representation of their constituent genes. Previous approaches to gene circuit design, such as CNN and RNN architectures, are unable to capture semantic dependencies in long contexts, as required in most real-world applications of synthetic biology. For instance, RNN models (LSTM, GRU), although able to learn long-term dependencies, greatly suffer from vanishing gradient and low-efficiency problem when they sequentially process past states and compresses contextual information into a bottleneck with long input sequences. In other words, these architectures are not equipped with the necessary attention mechanisms to follow a long chain of genes with thousands of tokens. To address the above-mentioned limitations, a transformer model was built in this work as a variation to the existing DNA Bidirectional Encoder Representations from Transformers (DNABERT) model. It is shown that the proposed transformer is capable of capturing contextual information from long input sequences with an attention mechanism. In previous works on genetic circuit design, the traditional approaches to classification and regression, such as Random Forrest, Support Vector Machine, and Artificial Neural Networks, were able to achieve reasonably high R2 accuracy levels of 0.95 to 0.97. However, the transformer model utilized in this work, with its attention-based mechanism, was able to achieve a perfect accuracy level of 100%. Further, it is demonstrated that the efficiency of the transformer-based gene expression classifier is not dependent on the presence of large amounts of training examples, which may be difficult to compile in many real-world gene circuit designs.

Keywords: machine learning, classification and regression, gene circuit design, bidirectional transformers

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Authors: J. Nyenhuis, J. Drelich

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This research study was designed to determine if food crops could be bio-fortified with micro-nutrients by growing sprouts on mineral fortified fiber mats. Diets high in processed foods have been found to lack essential micro-nutrients for optimum human development and overall health. Some micro-nutrients such as copper (Cu) have been found to enhance the inflammatory response through its oxidative functions, thereby having a role in cardiovascular disease (CVD), metabolic syndrome (MetS), diabetes and related complications. Recycled cellulose fibers and clay saturated with micro-nutrient ions can be converted to a novel mineral-metal hybrid material in which the fiber mat becomes a carrier of essential micro-nutrients. The reduction of ionic to metallic copper was accomplished using hydrogen at temperatures ranging from 400o to 600oC. Copper particles with diameters ranging from ~1 to 400-500 nm reside on the recycled fibers that make up the mats. Seeds purchased from a commercial, organic supplier were germinated on the specially engineered cellulose fiber mats that incorporated w10 wt% clay fillers saturated with either copper particles or ionic copper. After the appearance of the first leaves, the sprouts were dehydrated and analyzed for Cu content. Nutrient analysis showed 1.5 to 1.6 increase in Cu of the sprouts grown on the fiber mats with copper particles, and 2.3 to 2.5 increase on mats with ionic copper as compared to the control samples. The antibacterial properties of materials saturated with copper ions at room temperature and at temperatures up to 400°C have been verified with halo method tests against Escherichia Coli in previous studies. E. coli is a known pathogenic risk in sprout production. Copper exhibits excellent antibacterial properties when tested on S. aureus, a pathogenic gram-positive bacterium. This has also been confirmed for the fiber-copper hybrid material in this study. This study illustrates the potential for the use of engineered mats as a viable way to increase the micro-nutrient composition of locally-grown food crops and the need for additional research to determine the uptake, nutritional implications and risks of micro-nutrient bio-fortification.

Keywords: bio-fortification, copper nutrient analysis, micro-nutrient uptake, sprouts and mineral-fortified mats

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Authors: Asaf Toker

Abstract:

Matricelf developed a technology that enables the production of autologous engineered tissue composed of matrix and cells derived from patients Omentum biopsy. The platform showed remarkable pre-clinical results for several medical conditions. The company recently licensed the technology that enabled scientist at Tel Aviv university that 3D printed a human heart from human cells and matrix for the first time in human history. The company plans to conduct its first human clinical trial for Acute Spinal Cord Injury (SCI) early in 2023.

Keywords: tissue engineering, regenerative medicine, spinal Cord Injury, autologous implants, iPSC

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Authors: M. Naresh, Pratik Chaturvedi, Srishti Yadav, Varun Dutt, K. V. Uday

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Changes in the Earth’s climate are likely to increase natural hazards such as drought, floods, earthquakes, landslides, etc. The present study focusing on to early warning systems (EWS) of landslides, major issues in Himalayan region without prominence to deforestation, encroachments and un-engineered cutting of slopes and reforming for infrastructural purposes. EWS can be depicted by conducting a series of flume tests using micro-electro mechanical systems sensors data after reaching threshold values under controlled laboratory conditions. Based on the threshold value database, an alert will be sent via SMS.

Keywords: slope-instability, flume test, sensors, early warning system

Procedia PDF Downloads 240

Authors: Kamran Jawed, Anu Jose Mattam, Zia Fatma, Saima Wajid, Malik Z. Abdin, Syed Shams Yazdani

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Worldwide demand of natural and sustainable fuels and chemicals have encouraged researchers to develop microbial platform for synthesis of short chain fatty acids as they are useful precursors to replace petroleum-based fuels and chemicals. In this study, we evaluated the role of fatty acid synthesis and β-oxidation cycle of Escherichia coli to produce butyric acid, a 4-carbon short chain fatty acid, with the help of three thioesterases, i.e., TesAT from Anaerococcus tetradius, TesBF from Bryantella formatexigens and TesBT from Bacteroides thetaiotaomicron. We found that E. coli strain transformed with gene for TesBT and grown in presence of 8 g/L glucose produced maximum butyric acid titer at 1.46 g/L, followed by that of TesBF at 0.85 g/L and TesAT at 0.12 g/L, indicating that these thioesterases were efficiently converting short chain fatty acyl-ACP intermediate of fatty acid synthesis pathway into the corresponding acid. The titer of butyric acid varied significantly depending upon the plasmid copy number and strain genotype. Deletion of genes for fatty acyl-CoA synthetase and acyl-CoA dehydrogenase, which are involved in initiating the fatty acid degradation cycle, and overexpression of FadR, which is a dual transcriptional regulator and exerts negative control over fatty acid degradation pathway, reduced up to 30% of butyric acid titer. This observation suggested that β-oxidation pathway is working synergistically with fatty acid synthesis pathway in production of butyric acid. Moreover, accelerating the fatty acid elongation cycle by overexpressing acetyl-CoA carboxyltransferase (Acc) and 3-hydroxy-acyl-ACP dehydratase (FabZ) or by deleting FabR, the transcription suppressor of elongation, did not improve the butyric acid titer, rather favored the long chain fatty acid production. Finally, a balance between cell growth and butyric acid production was achieved with the use of phosphorous limited growth medium and 14.3 g/L butyric acid, and 17.5 g/L total free fatty acids (FFAs) titer was achieved during fed-batch cultivation. We have engineered an E. coli strain which utilizes the intermediate of both fatty acid synthesis and degradation pathway, i.e. butyryl-ACP and -CoA, to produce butyric acid from glucose. The strategy used in this study resulted in highest reported titers of butyric acid and FFAs in engineered E. coli.

Keywords: butenoic acid, butyric acid, Escherichia coli, fed-batch fermentation, short chain fatty acids, thioesterase

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Authors: Lucy Horrobin, Rebecca Fores

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Background: Muay Thai is an increasingly popular combat sport worldwide. Further academic research in the sport will contribute to its professional development. This research sought to produce normative data in relation to the physical and physiological characteristics of professional Muay Thai boxers, as, currently no such data exists. The ultimate aim being to inform appropriate training programs and to facilitate coaching. Methods: N = 9 professional, adult, male Muay Thai boxers were assessed for the following anthropometric, physical and physiological characteristics, using validated methods of assessment: body fat, hamstring flexibility, maximal dynamic upper body strength, lower limb peak power, upper body muscular endurance and aerobic capacity. Raw data scores were analysed for mean, range and SD and where applicable were expressed relative to body mass (BM). Results: Results showed similar characteristics to those found in other combat sports. Low percentages of body fat (mean±SD) 8.54 ± 1.16 allow for optimal power to weight ratios. Highly developed aerobic capacity (mean ±SD) 61.56 ± 5.13 ml.min.kg facilitate recovery and power maintenance throughout bouts. Lower limb peak power output values of (mean ± SD) 12.60 ± 2.09 W/kg indicate that Muay Thai boxers are amongst the most powerful of combat sport athletes. However, maximal dynamic upper body strength scores of (mean±SD) 1.14 kg/kg ± 0.18 were in only the 60th percentile of normative data for the general population and muscular endurance scores (mean±SD) 31.55 ± 11.95 and flexibility scores (mean±SD) 19.55 ± 11.89 cm expressed wide standard deviation. These results might suggest that these characteristics are insignificant in Muay Thai or under-developed, perhaps due to deficient training programs. Implications: This research provides the first normative data of physical and physiological characteristics of Muay Thai boxers. The findings of this study would aid trainers and coaches when designing effective evidence-based training programs. Furthermore, it provides a foundation for further research relating to physiology in Muay Thai. Areas of further study could be determining the physiological demands of a full rules bout and the effects of evidence-based training programs on performance.

Keywords: fitness testing, Muay Thai, physiology, strength and conditioning

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Authors: Som Nath Sachdeva, Vanita Aggarwal, S. M. Gupta

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Use of concrete paver blocks is becoming increasingly popular. They are used for paving of approaches, paths and parking areas including their application in pre-engineered buildings. This paper discusses the results of an experimental study conducted on Fly Ash Concrete with the aim to report its suitability for concrete paver blocks. In this study, the effect of varying proportions of fly ash, 20 % to 40 %, on compressive strength and flexural strength of concrete has been evaluated. The mix designs studied are M-30, M-35, M-40 and M-50. It is observed that all the fly ash based mixes are able to achieve the required compressive and flexural strengths. In comparison to control mixes, the compressive and flexural strengths of the fly ash based mixes are found to be slightly less at 7 days and 28 days and a little more at 90 days.

Keywords: fly ash concrete, paver blocks, compressive, flexural strength

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Authors: Pedro Llanos, Diego García

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Spaceflight is considered the last frontier in terms of science, technology, and engineering. But it is also the next frontier in terms of human physiology and performance. After more than 200,000 years humans have evolved under earth’s gravity and atmospheric conditions, spaceflight poses environmental stresses for which human physiology is not adapted. Hypoxia, accelerations, and radiation are among such stressors, our research involves suborbital flights aiming to develop effective countermeasures in order to assure sustainable human space presence. The physiologic baseline of spaceflight participants is subject to great variability driven by age, gender, fitness, and metabolic reserve. The objective of the present study is to characterize different physiologic variables in a population of STEM practitioners during an aerobatic flight. Cardiovascular and pulmonary responses were determined in Science Astronaut Candidates (SACs) during unusual attitude aerobatic flight indoctrination. Physiologic data recordings from 20 subjects participating in high-G flight training were analyzed. These recordings were registered by wearable sensor-vest that monitored electrocardiographic tracings (ECGs), signs of dysrhythmias or other electric disturbances during all the flight. The same cardiovascular parameters were also collected approximately 10 min pre-flight, during each high-G/unusual attitude maneuver and 10 min after the flights. The ratio (pre-flight/in-flight/post-flight) of the cardiovascular responses was calculated for comparison of inter-individual differences. The resulting tracings depicting the cardiovascular responses of the subjects were compared against the G-loads (Gs) during the aerobatic flights to analyze cardiovascular variability aspects and fluid/pressure shifts due to the high Gs. In-flight ECG revealed cardiac variability patterns associated with rapid Gs onset in terms of reduced heart rate (HR) and some scattered dysrhythmic patterns (15% premature ventricular contractions-type) that were considered as triggered physiological responses to high-G/unusual attitude training and some were considered as instrument artifact. Variation events were observed in subjects during the +Gz and –Gz maneuvers and these may be due to preload and afterload, sudden shift. Our data reveal that aerobatic flight influenced the breathing rate of the subject, due in part by the various levels of energy expenditure due to the increased use of muscle work during these aerobatic maneuvers. Noteworthy was the high heterogeneity in the different physiological responses among a relatively small group of SACs exposed to similar aerobatic flights with similar Gs exposures. The cardiovascular responses clearly demonstrated that SACs were subjected to significant flight stress. Routine ECG monitoring during high-G/unusual attitude flight training is recommended to capture pathology underlying dangerous dysrhythmias in suborbital flight safety. More research is currently being conducted to further facilitate the development of robust medical screening, medical risk assessment approaches, and suborbital flight training in the context of the evolving commercial human suborbital spaceflight industry. A more mature and integrative medical assessment method is required to understand the physiology state and response variability among highly diverse populations of prospective suborbital flight participants.

Keywords: g force, aerobatic maneuvers, suborbital flight, hypoxia, commercial astronauts

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Authors: Filipa Cruz, Grecia P. Matos

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An Inexhaustible Will of Infinite is ongoing practice-based research focused on a psychophysiological conception of body and on the creative will that seeks to examine the possibility of art being simultaneously a pacifier and an intensifier in a physiological artistic production. This is a study where philosophy and art converge in a commentary on the affection of the concept of will to power in the art world through Nietzsche’s commentaries, through the analysis of case studies and a reflection arising from artistic practice. Through Nietzsche, it is sought to compare concepts that communicate with the artistic practice since creation is an intensification and engenders perspectives. It is also a practice highly embedded in the body, in the non-verbal, in the physiology of art and in the coexistence between the sensorial and the thought. It is questioned if the physiology of art could be thought of as a thinking-feeling with no primacy of the thought over the sensorial. Art as a manifestation of the will to power participates in a comprehension of the world. In this article, art is taken as a privileged way of communication – implicating corporeal-sensorial-conceptual – and of connection between humans. Problematized is the dream and the drunkenness as intensifications and expressions of life’s comprehension. Therefore, art is perceived as suggestion and invention, where the artistic intoxication breaks limits in the experience of life, and the artist, dominated by creative forces, claims, orders, obeys, proclaims love for life. The intention is also to consider how one can start from pain to create and how one can generate new and endless artistic forms through nightmares, daydreams, impulses, intoxication, enhancement, intensification in a plurality of subjects and matters. It is taken into consideration the fact that artistic creation is something that is intensified corporeally, expanded, continuously generated and acting on bodies. It is inextinguishable and a constant movement intertwining Apollonian and Dionysian instincts of destruction and creation of new forms. The concept of love also appears associated with conquering, that, in a process of intensification and drunkenness, impels the artist to generate and to transform matter. Just like a love relationship, love in Nietzsche requires time, patience, effort, courage, conquest, seduction, obedience, and command, potentiating the amplification of knowledge of the other / the world. Interlacing Nietzsche's philosophy, not with Modern Art, but with Contemporary Art, it is argued that intoxication, will to power (strongly connected with the creative will) and love still have a place in the artistic production as creative agents.

Keywords: artistic creation, body, intensification, psychophysiology, will to power

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Authors: Inna Kornienko, Svetlana Guryeva, Natalia Danilova, Elena Petersen

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Drug toxicity often goes undetected until clinical trials, the most expensive and dangerous phase of drug development. Both human cell culture and animal studies have limitations that cannot be overcome by improvements in drug testing protocols. Tissue engineering is an emerging alternative approach to creating models of human malignant tumors for experimental oncology, personalized medicine, and drug discovery studies. This new generation of bioengineered tumors provides an opportunity to control and explore the role of every component of the model system including cell populations, supportive scaffolds, and signaling molecules. An area that could greatly benefit from these models is cancer research. Recent advances in tissue engineering demonstrated that decellularized tissue is an excellent scaffold for tissue engineering. Decellularization of donor organs such as heart, liver, and lung can provide an acellular, naturally occurring three-dimensional biologic scaffold material that can then be seeded with selected cell populations. Preliminary studies in animal models have provided encouraging results for the proof of concept. Decellularized Organs preserve organ microenvironment, which is critical for cancer metastasis. Utilizing 3D tumor models results greater proximity of cell culture morphological characteristics in a model to its in vivo counterpart, allows more accurate simulation of the processes within a functioning tumor and its pathogenesis. 3D models allow study of migration processes and cell proliferation with higher reliability as well. Moreover, cancer cells in a 3D model bear closer resemblance to living conditions in terms of gene expression, cell surface receptor expression, and signaling. 2D cell monolayers do not provide the geometrical and mechanical cues of tissues in vivo and are, therefore, not suitable to accurately predict the responses of living organisms. 3D models can provide several levels of complexity from simple monocultures of cancer cell lines in liquid environment comprised of oxygen and nutrient gradients and cell-cell interaction to more advanced models, which include co-culturing with other cell types, such as endothelial and immune cells. Following this reasoning, spheroids cultivated from one or multiple patient-derived cell lines can be utilized to seed the matrix rather than monolayer cells. This approach furthers the progress towards personalized medicine. As an initial step to create a new ex vivo tissue engineered model of a cancer tumor, optimized protocols have been designed to obtain organ-specific acellular matrices and evaluate their potential as tissue engineered scaffolds for cultures of normal and tumor cells. Decellularized biomatrix was prepared from animals’ kidneys, urethra, lungs, heart, and liver by two decellularization methods: perfusion in a bioreactor system and immersion-agitation on an orbital shaker with the use of various detergents (SDS, Triton X-100) in different concentrations and freezing. Acellular scaffolds and tissue engineered constructs have been characterized and compared using morphological methods. Models using decellularized matrix have certain advantages, such as maintaining native extracellular matrix properties and biomimetic microenvironment for cancer cells; compatibility with multiple cell types for cell culture and drug screening; utilization to culture patient-derived cells in vitro to evaluate different anticancer therapeutics for developing personalized medicines.

Keywords: 3D models, decellularization, drug discovery, drug toxicity, scaffolds, spheroids, tissue engineering

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Authors: Sara R. Knigge, Sugat R. Tuladhar, Hans-Klaus HöFfler, Tobias Schilling, Tim Kaufeld, Axel Haverich

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Tissue engineered (TE) heart valves based on degradable electrospun fiber scaffold represent a promising approach to overcome the known limitations of mechanical or biological prostheses. But the mechanical stress in the high-pressure system of the human circulation is a severe challenge for the delicate materials. Hence, the prediction of the scaffolds` in vivo degradation kinetics must be as accurate as possible to prevent fatal events in future animal or even clinical trials. Therefore, this study investigates whether long-term testing in full blood provides more meaningful results regarding the degradation behavior than conventional tests in simulated body fluids (SBF) or Phosphate Buffered Saline (PBS). Fiber mats were produced from a polycaprolactone (PCL)/tetrafluoroethylene solution by electrospinning. The morphology of the fiber mats was characterized via scanning electron microscopy (SEM). A maximum physiological degradation environment utilizing a test set-up with porcine full blood was established. The set-up consists of a reaction vessel, an oxygenator unit, and a roller pump. The blood parameters (pO2, pCO2, temperature, and pH) were monitored with an online test system. All tests were also carried out in the test circuit with SBF and PBS to compare conventional degradation media with the novel full blood setting. The polymer's degradation is quantified by SEM picture analysis, differential scanning calorimetry (DSC), and Raman spectroscopy. Tensile and cyclic loading tests were performed to evaluate the mechanical integrity of the scaffold. Preliminary results indicate that PCL degraded slower in full blood than in SBF and PBS. The uptake of water is more pronounced in the full blood group. Also, PCL preserved its mechanical integrity longer when degraded in full blood. Protein absorption increased during the degradation process. Red blood cells, platelets, and their aggregates adhered on the PCL. Presumably, the degradation led to a more hydrophilic polymeric surface which promoted the protein adsorption and the blood cell adhesion. Testing degradable implants in full blood allows for developing more reliable scaffold materials in the future. Material tests in small and large animal trials thereby can be focused on testing candidates that have proven to function well in an in-vivo-like setting.

Keywords: Electrospun scaffold, full blood degradation test, long-term polymer degradation, tissue engineered aortic heart valve

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Authors: Yadnyavalkya Patil, Ravi Gooneratne, Xiang-Hong Ju

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Heat stress (HS) negatively affects the physiology of pigs. In this study, 6 pigs will be subjected to temperatures of 35 ± 2℃ for 12 hrs/day for a duration of 21 days. The changes in the gastrointestinal tract (GIT) microbiota will be observed by analyzing the freshly collected faeces on days 1, 3, 7, 14 and 21. The changes will be compared to faeces from a set of 6 control pigs kept simultaneously at temperatures of 26 ± 2℃ for the same duration of 21 days. Different types of stresses such a weaning have a detrimental effect on GIT microflora. Similarly, HS is expected to have a harmful effect on the microbial diversity of the GIT. How these changes affect the immune system of the pigs will be studied and therapeutics to reduce the negative effects of HS will be developed.

Keywords: GIT microbiota, heat stress, immune system, therapeutics

Procedia PDF Downloads 182

Authors: Osama Moustafa Zayed

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Aim of the work: To evaluate the effectiveness of venous glucose, levels of serum lactate and base deficit in polytraumatized patients as simple parameters to predict the mortality in these patients. Compared to the predictive value of Trauma and injury severity (TRISS) and Acute Physiology And Chronic Health Evaluation IV (APACHE IV). Introduction: Trauma is a serious global health problem, accounting for approximately one in 10 deaths worldwide. Trauma accounts for 5 million deaths per year. Prediction of mortality in trauma patients is an important part of trauma care. Several trauma scores have been devised to predict injury severity and risk of mortality. The trauma and injury severity score (TRISS) was most common used. Regardless of the accuracy of trauma scores, is based on an anatomical description of every injury and cannot be assigned to the patients until a full diagnostic procedure has been performed. So we hypothesized that alterations in admission glucose, lactate levels and base deficit would be an early and easy rapid predictor of mortality. Patient and Method: a comparative cross-sectional study. 282 Polytraumatized patients attended to the Emergency Department(ED) of the Suez Canal university Hospital constituted. The period from 1/1/2012 to 1/4/2013 was included. Results: We found that the best cut off value of TRISS probability of survival score for prediction of mortality among poly-traumatized patients is = 90, with 77% sensitivity and 89% specificity using area under the ROC curve (0.89) at (95%CI). APACHE IV demonstrated 67% sensitivity and 95% specificity at 95% CI at cut off point 99. The best cutoff value of Random Blood Sugar (RBS) for prediction of mortality was>140 mg/dl, with 89%, sensitivity, 49% specificity. The best cut off value of base deficit for prediction of mortality was less than -5.6 with 64% sensitivity, 93% specificity. The best cutoff point of lactate for prediction of mortality was > 2.6 mmol/L with 92%, sensitivity, 42% specificity. Conclusion: According to our results from all evaluated predictors of mortality (laboratory and scores) and mortality based on the estimated cutoff values using ROC curves analysis, the highest risk of mortality was found using a cutoff value of 90 in TRISS score while with laboratory parameters the highest risk of mortality was with serum lactate > 2.6 . Although that all of the three parameter are accurate in predicting mortality in poly-traumatized patients and near with each other, as in serum lactate the area under the curve 0.82, in BD 0.79 and 0.77 in RBS.

Keywords: APACHE IV, emergency department, polytraumatized patients, serum lactate

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Authors: Prince Vivek, Vijay Kumar Bharti, Deepak Kumar, Rohit Kumar, Kapil Nehra, Dhananjay Singh, Om Prakash Chaurasia, Bhuvnesh Kumar

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High altitude native people still rely heavily on animal transport for logistic support at eastern and northern Himalayas regions. The prevalent mountainous terrains and rugged region are not suitable for the motorized vehicle to use in logistic transport. Therefore, people required high endurance pack animals for load carrying and riding. So far to the best of our knowledge, no studies have been taken to evaluate the effect of loads on the physiology of ponies in high altitude region. So, in this view, we evaluated variation in physiological, hematological, biochemical, and cytokines indices of Zanskar ponies during load carrying at high altitude. Total twelve (12) of Zanskar ponies, mare, age 4-6 years selected for this study, Feed was offered at 2% of body weight, and water ad libitum. Ponies were divided into three groups; group-A (without load), group-B (60 kg), and group-C (80 kg) of backpack loads. The track was very narrow and slippery with gravel, uneven with a rocky surface and has a steep gradient of 4 km uphill at altitude 3291 to 3500m. When we evaluate these parameters, it is understood that the heart rate, pulse rate, and respiration rate was significantly increased in 80 kg group among the three groups. The hematology parameters viz. hemoglobin significantly increased in 80 kg group on 1st day after load carrying among the three groups which was followed by control and 60 kg whereas, PCV, lymphocytes, monocytes percentage significantly increased however, ESR and eosinophil % significantly decreased in 80 kg group after load carrying on 7th day after load carrying among the three groups which were followed by control and 60 kg group. In biochemical parameters viz. LA, LDH, TP, hexokinase (HK), cortisol (CORT), T3, GPx, FRAP and IL-6 significantly increased in 80 kg group on the 7th day after load carrying among the three groups which were followed by control and 60 kg group. The ALT, ALB, GLB, UR, and UA significantly increased in 80 kg group on the 7th day before and after load carrying among the three groups which were followed by control and 60 kg group. The CRT, AST, and CK-MB were significantly increased in 80 kg group on the 1st and 7th day after load carrying among the three groups which were followed by control and 60 kg group. It has been concluded that, heart rate, respiration rate, hematological indices like PCV, lymphocytes, monocytes, Hb and ESR, biochemical indices like lactic acid, LDH, TP, HK, CORT, T3, ALT, AST and CRT, ALB, GLB, UR, UA, GPx, FRAP and IL-6 are important biomarkers to assess effect of load on animal physiology and endurance. Further, this result has revealed the strong correlation of change in biomarkers level with performance in ponies during load carry. Hence, these parameters might be used for the performance of endurance of Zanskar ponies in the high mountain region.

Keywords: biochemical, endurance, high altitude, load, ponies

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Authors: Genevieve Pugh, Johnathan Burns, Stefan Howorka

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Single-molecule sensing via protein nanopores has achieved a step-change in portable and label-free DNA sequencing. However, protein pores of both natural or engineered origin are not able to produce the tunable diameters needed for effective protein sensing. Here, we describe a generic strategy to build synthetic DNA nanopores that are wide enough to accommodate folded protein. The pores are composed of interlinked DNA duplexes and carry lipid anchors to achieve the required membrane insertion. Our demonstrator pore has a contiguous cross-sectional channel area of 50 nm2 which is 6-times larger than the largest protein pore. Consequently, transport of folded protein across bilayers is possible. The modular design is amenable for different pore dimensions and can be adapted for protein sensing or to create molecular gates in synthetic biology.

Keywords: biosensing, DNA nanotechnology, DNA origami, nanopore sensing

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Authors: Mohammed Fayez Al Rez

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Nowadays, there is an unmet clinical need for new small-diameter vascular grafts to overcome the drawbacks of traditional methods used for treatment of widespread cardiovascular diseases. Vascular tissue engineering (VTE) is a promising approach that can be utilized to develop viable vascular grafts by in vitro seeding of functional cells onto a scaffold allowing them to attach, proliferate and differentiate. To achieve this purpose, the scaffold should provide cells with the initial necessary extracellular matrix environment and structure until being able to reconstruct the required vascular tissue. Therefore, producing scaffolds with suitable features is crucial for guiding cells properly to develop the desired tissue-engineered vascular grafts for clinical applications. The main objective of this work is fabrication and characterization of tubular small-diameter ( < 6 mm) bilayered scaffolds for VTE. The scaffolds were prepared via mixing electrospinning approach of biodegradable poly(ε-caprolactone) (PCL) polymer – due to its favorable physicochemical properties – to mimic the natural environment-extracellular matrix. Firstly, tubular nanofibrous construct with inner diameter of 3, 4 or 5 mm was electrospun as inner layer, and secondly, microfibrous construct was electrospun as outer layer directly on the first produced inner layer. To improve the biological properties of PCL, a group of the electrospun scaffolds was immersed in type-1 collagen solution. The morphology and structure of the resulting fibrous scaffolds were investigated by scanning electron microscope. The electrospun nanofibrous inner layer contained fibers measuring 219±35 nm in diameter, while the electrospun microfibrous outer layer contained fibers measuring 1011 ± 150 nm. Furthermore, mechanical, thermal and physical tests were conducted with both electrospun bilayered scaffold types where revealed improved properties. Biological investigations using endothelial, smooth muscle and fibroblast cell line showed good biocompatibility of both tested electrospun scaffolds. Better attachment and proliferation were obviously found when cells were cultured on the scaffolds immersed with collagen due to increasing the hydrophilicity of the PCL. The easy, inexpensive and versatile electrospinning approach used in this work was able to successfully produce double layered tubular elastic structures containing both nanofibers and microfibers to imitate the native vascular structure. The PCL – as a suitable and approved biomaterial for many biomedical and tissue engineering applications – can ensure favorable mechanical properties of scaffolds used for VTE. The VTE approach using electrospun bilayered scaffolds offers optimal solutions and holds significant promises for treatment of many cardiovascular diseases.

Keywords: electrospinning, poly(ε-caprolactone) (PCL), tissue-engineered vascular graft, tubular bilayered scaffolds, vascular cells

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Authors: Naresh Radaliyagoda, Nigel M. Jennett, Rong Lan, David Parfitt

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A series of length scale engineered multilayer material with temperature robust mechanical properties has been suggested. A range of polycrystalline copper sub-layers with the thickness varying from 1 to 25μm and buried in between two nickel layers was produced using electrodeposition dual bath technique. The structure of the multilayers was characterized using Electron Backscatter Diffraction and Scanning Electron Microscope. The interface effect on the hardness and elastic modulus was tested using Nano-indentation. Results of the grain size and layer thickness measurements, and indentation hardness have been compared. It is found that there is a combined length scale effect that improves mechanical properties in Ni/Cu multilayer structures.

Keywords: nano-indentation, size effect, multilayers, electrodeposition

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Authors: M. Yusuf

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The recent discovery of the phenomenon of RNA interference has led to its application in various aspects of plant improvement. Crops can be modified by engineering novel RNA interference pathways that create small RNA molecules to alter gene expression in crops or plant pests. RNA interference can generate new crop quality traits or provide protection against insects, nematodes and pathogens without introducing new proteins into food and feed products. This is an advantage in contrast with conventional procedures of gene transfer. RNA interference has been used to develop crop varieties resistant to diseases, pathogens and insects. Male sterility has been engineered in plants using RNA interference. Better quality crops have been developed through the application of RNA interference etc. The objective of this paper is to highlight the application of RNA interference in crop improvement and to project its potential future use to solve problems of agricultural production in relation to plant breeding.

Keywords: RNA interference, application, crop Improvement, agricultural production

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Authors: Md. Asaduzzaman

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A Silicon-on-insulator (SOI) perfectly vertical fibre-to-chip grating coupler is proposed and designed based on engineered subwavelength structures. The high directionality of the coupler is achieved by implementing step gratings to realize asymmetric diffraction and by applying effective index variation with auxiliary ultra-subwavelength gratings. The proposed structure is numerically analysed by using two-dimensional Finite Difference Time Domain (2D FDTD) method and achieves 96% (-0.2 dB) coupling efficiency and 39 nm 1-dB bandwidth. This highly efficient GC is necessary for applications where coupling efficiency between the optical fibre and nanophotonics waveguide is critically important, for instance, experiments of the quantum photonics integrated circuits. Such efficient and broadband perfectly vertical grating couplers are also significantly advantageous in highly dense photonic packaging.

Keywords: diffraction grating, FDTD, grating couplers, nanophotonic

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Authors: Camille Paldi

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The tense dichotomy between Shari’ah compliance and the economic goals of an Islamic Bank produces a proliferation of reverse engineered products, which are barely in compliance with Islamic law. The result is basically a hybrid conventional banking system with conventional products in Islamic disguise using Arabic and Islamic terminology. Many Islamic financial professionals and academics advocate for the use of conventional products and devices despite their non-Shari’ah compliance based on commercial necessity and the need to compete. However, this dangerous trend will lead to the demise of the Islamic finance industry. Rather than thoughtlessly following conventional products and practice, Islamic finance professionals should delve into the Shari’ah to find the answers to the current Islamic banking conundrum and lead the industry on the right path of developing Shari’ah based products and using Shari’ah devices to hedge risk.

Keywords: Islamic banking, Shari'ah, finance, investment

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Authors: Temesgen Geremew

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Functionalized nanoparticles have emerged as a revolutionary class of materials with immense potential in various biomedical applications. These engineered nanoparticles possess unique properties tailored to interact with biological systems, offering unprecedented opportunities in drug delivery, imaging, diagnostics, and therapy. This research delves into the design, synthesis, and characterization of functionalized nanoparticles for targeted biomedical applications. The primary focus lies on developing nanoparticles with precisely controlled size, surface chemistry, and biocompatibility for specific medical purposes. The research will also explore the crucial interaction of these nanoparticles with biological systems, encompassing cellular uptake, biodistribution, and potential toxicity evaluation. The successful development of functionalized nanoparticles holds the promise to revolutionize various aspects of healthcare. This research aspires to contribute significantly to this advancement by providing valuable insights into the design and application of these versatile materials within the ever-evolving field of biomedicine.

Keywords: nanoparticles, biomedicals, cancer, biocompatibility

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Authors: M. S. Sukumar

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Our objective is to develop a full-fledged system for exploring and studying nature of fossils and to extend this to underwater archaeology and mineral mapping. This includes aerial surveying, imaging techniques, artefact extraction and spectrum analysing techniques. These techniques help in regular monitoring of fossils and also the sensing system. The ROV was designed to complete several tasks which simulate collecting data and samples. Given the time constraints, the ROV was engineered for efficiency and speed in performing tasks. Its other major design consideration was modularity, allowing the team to distribute the building process, to easily test systems as they were completed and troubleshoot and replace systems as necessary. Our design itself had several challenges of on-board waterproofed sensor mounting, waterproofing of motors, ROV stability criteria, camera mounting and hydrophone sound acquisition.

Keywords: remotely operated vehicle (ROV) dragonair, underwater archaeology, full-fledged system, aerial imaging and detection

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Authors: Natalia Efremova, Sergey Tarasenko

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We propose a novel neural network architecture for visual saliency detections, which utilizes neuro physiologically plausible mechanisms for extraction of salient regions. The model has been significantly inspired by recent findings from neuro physiology and aimed to simulate the bottom-up processes of human selective attention. Two types of features were analyzed: color and direction of maximum variance. The mechanism we employ for processing those features is PCA, implemented by means of normalized Hebbian learning and the waves of spikes. To evaluate performance of our model we have conducted psychological experiment. Comparison of simulation results with those of experiment indicates good performance of our model.

Keywords: neural network models, visual saliency detection, normalized Hebbian learning, Oja's rule, psychological experiment

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Authors: Muneerah Jeludin

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The vibrated column has been widely used over the last three decades to improve the performance of soft ground and engineered compacted fill. The main reason for adopting this technique is that it is economically viable and environmental friendly. The performance of granular column with regards to bearing capacity has been well documented; however, information regarding the settlement behavior of granular columns is still limited. This paper aims to address the findings from a laboratory model study in terms of its settlement improvement. A 300 mm diameter and 400 mm high kaolin clay model was used in this investigation. Columns of various heights were installed in the clay bed using replacement method. The results in relation to load sharing mechanism between the column and surrounding clay just under the footing indicated that in short column, the available shaft resistance was not significant and introduces a potential for end braing failure as opposed to bulging failure in long columns. The settlement improvement factor corroborates well with field observations.

Keywords: ground improvement, model test, reinforced soil, foundation

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Authors: Yuriko Takayama, Norihiro Kato

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Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

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