Search results for: protein distance maps
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4936

Search results for: protein distance maps

286 Effect of Minimalist Footwear on Running Economy Following Exercise-Induced Fatigue

Authors: Jason Blair, Adeboye Adebayo, Mohamed Saad, Jeannette M. Byrne, Fabien A. Basset

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Running economy is a key physiological parameter of an individual’s running efficacy and a valid tool for predicting performance outcomes. Of the many factors known to influence running economy (RE), footwear certainly plays a role owing to its characteristics that vary substantially from model to model. Although minimalist footwear is believed to enhance RE and thereby endurance performance, conclusive research reports are scarce. Indeed, debates remain as to which footwear characteristics most alter RE. The purposes of this study were, therefore, two-fold: (a) to determine whether wearing minimalist shoes results in better RE compared to shod and to identify relationships with kinematic and muscle activation patterns; (b) to determine whether changes in RE with minimalist shoes are still evident following a fatiguing bout of exercise. Well-trained male distance runners (n=10; 29.0 ± 7.5 yrs; 71.0 ± 4.8 kg; 176.3 ± 6.5 cm) partook first in a maximal O₂ uptake determination test (VO₂ₘₐₓ = 61.6 ± 7.3 ml min⁻¹ kg⁻¹) 7 days prior to the experimental sessions. Second, in a fully randomized fashion, an RE test consisting of three 8-min treadmill runs in shod and minimalist footwear were performed prior to and following exercise induced fatigue (EIF). The minimalist and shod conditions were tested with a minimum of 7-day wash-out period between conditions. The RE bouts, interspaced by 2-min rest periods, were run at 2.79, 3.33, and 3.89 m s⁻¹ with a 1% grade. EIF consisted of 7 times 1000 m at 94-97% VO₂ₘₐₓ interspaced with 3-min recovery. Cardiorespiratory, electromyography (EMG), kinematics, rate of perceived exertion (RPE) and blood lactate were measured throughout the experimental sessions. A significant main speed effect on RE (p=0.001) and stride frequency (SF) (p=0.001) was observed. The pairwise comparisons showed that running at 2.79 m s⁻¹ was less economic compared to 3.33, and 3.89 m s⁻¹ (3.56 ± 0.38, 3.41 ± 0.45, 3.40 ± 0.45 ml O₂ kg⁻¹ km⁻¹; respectively) and that SF increased as a function of speed (79 ± 5, 82 ± 5, 84 ± 5 strides min⁻¹). Further, EMG analyses revealed that root mean square EMG significantly increased as a function of speed for all muscles (Biceps femoris, Gluteus maximus, Gastrocnemius, Tibialis anterior, Vastus lateralis). During EIF, the statistical analysis revealed a significant main effect of time on lactate production (from 2.7 ± 5.7 to 11.2 ± 6.2 mmol L⁻¹), RPE scores (from 7.6 ± 4.0 to 18.4 ± 2.7) and peak HR (from 171 ± 30 to 181 ± 20 bpm), expect for the recovery period. Surprisingly, a significant main footwear effect was observed on running speed during intervals (p=0.041). Participants ran faster with minimalist shoes compared to shod (3:24 ± 0:44 min [95%CI: 3:14-3:34] vs. 3:30 ± 0:47 min [95%CI: 3:19-3:41]). Although EIF altered lactate production and RPE scores, no other effect was noticeable on RE, EMG, and SF pre- and post-EIF, except for the expected speed effect. The significant footwear effect on running speed during EIF was unforeseen but could be due to shoe mass and/or heel-toe-drop differences. We also cannot discard the effect of speed on foot-strike pattern and therefore, running performance.

Keywords: exercise-induced fatigue, interval training, minimalist footwear, running economy

Procedia PDF Downloads 215
285 Bone Mineralization in Children with Wilson’s Disease

Authors: Shiamaa Eltantawy, Gihan Sobhy, Alif Alaam

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Wilson disease, or hepatolenticular degeneration, is an autosomal recessive disease that results in excess copper buildup in the body. It primarily affects the liver and basal ganglia of the brain, but it can affect other organ systems. Musculoskeletal abnormalities, including premature osteoarthritis, skeletal deformity, and pathological bone fractures, can occasionally be found in WD patients with a hepatic or neurologic type. The aim was to assess the prevalence of osteoporosis and osteopenia in Wilson’s disease patients. This case-control study was conducted on ninety children recruited from the inpatient ward and outpatient clinic of the Paediatric Hepatology, Gastroenterology, and Nutrition department of the National Liver Institute at Menofia University, aged from 1 to 18 years. Males were 49, and females were 41. Children were divided into three groups: (Group I) consisted of thirty patients with WD; (Group II) consisted of thirty patients with chronic liver disease other than WD; (Group III) consisted of thirty age- and sex-matched healthy The exclusion criteria were patients with hyperparathyroidism, hyperthyroidism, renal failure, Cushing's syndrome, and patients on certain drugs such as chemotherapy, anticonvulsants, or steroids. All patients were subjected to the following: 1- Full history-taking and clinical examination. 2-Laboratory investigations: (FBC,ALT,AST,serum albumin, total protein, total serum bilirubin,direct bilirubin,alkaline phosphatase, prothrombin time, serum critine,parathyroid hormone, serum calcium, serum phosphrus). 3-Bone mineral density (BMD, gm/cm2) values were measured by dual-energy X-ray absorptiometry (DEXA). The results revealed that there was a highly statistically significant difference between the three groups regarding the DEXA scan, and there was no statistically significant difference between groups I and II, but the WD group had the lowest bone mineral density. The WD group had a large number of cases of osteopenia and osteoporosis, but there was no statistically significant difference with the group II mean, while a high statistically significant difference was found when compared to group III. In the WD group, there were 20 patients with osteopenia, 4 patients with osteoporosis, and 6 patients who were normal. The percentages were 66.7%, 13.3%, and 20%, respectively. Therefore, the largest number of cases in the WD group had osteopenia. There was no statistically significant difference found between WD patients on different treatment regimens regarding DEXA scan results (Z-Score). There was no statistically significant difference found between patients in the WD group (normal, osteopenic, or osteoporotic) regarding phosphorus (mg/dL), but there was a highly statistically significant difference found between them regarding ionised Ca (mmol/L). Therefore, there was a decrease in bone mineral density when the Ca level was decreased. In summary, Wilson disease is associated with bone demineralization. The largest number of cases in the WD group in our study had osteopenia (66.7%). Different treatment regimens (zinc monotherapy, Artamin, and zinc) as well as different laboratory parameters have no effect on bone mineralization in WD cases. Decreased ionised Ca is associated with low BMD in WD patients. Children with WD should be investigated for BMD.

Keywords: wilson disease, Bone mineral density, liver disease, osteoporosis

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284 Potential of Water Purification of Turbid Surface Water Sources in Remote Arid and Semi-Arid Rural Areas of Rajasthan by Moringa Oleifera (Drumstick) Tree Seeds

Authors: Pomila Sharma

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Rajasthan is among regions with greatest climate sensitivity and lowest adaptive capabilities. In many parts of the Rajasthan surface water which can be highly turbid and contaminated with fecal coliform bacteria is used for drinking purposes. The majority rely almost exclusively upon traditional sources of highly turbid and untreated pathogenic surface water for their domestic water needs. In many parts of rural areas of Rajasthan, it is still difficult to obtain clean water, especially remote habitations with no groundwater due to quality issues or depletion and limited feasibility to connect with surface water schemes due to low density of population in these areas to justify large infrastructure investment. The most viable sources are rain water harvesting, community managed open wells, private wells, ponds and small-scale irrigation reservoirs have often been the main traditional sources of rural drinking water. Turbidity is conventionally removed by treating the water with expensive chemicals. This study has to investigate the use of crushed seeds from the tree Moringa oleifera (drumstick) as a natural alternative to conventional coagulant chemicals. The use of Moringa oleifera seed powder can produce potable water of higher quality than the original source. Moringa oleifera a native species of northern India, the tree is now grown extensively throughout the tropics and found in many countries of Africa, Asia & South America. The seeds of tree contains significant quantities of low molecular weight, water soluble proteins which carries the positive charge when the crushed seeds are added to water. This protein binds in raw water with negatively charged turbid water with bacteria, clay, algae, etc. Under proper mixing, these particles make flocks, which may be left to settle by gravity or be removed by filtration. Using Moringa oleifera as a replacement coagulation in such surface sources of arid and semi-arid areas can meet the need for water purification in remote places of Rajasthan state of India. The present study accesses to find out laboratory based investigation of the effect of seeds of Moringa tree on its coagulation effectiveness (purification) using turbid water samples of surface source of the Rajasthan state. In this study, moringa seed powder showed that filtering with seed powder may diminish water pollution and bacterial counts. Results showed Moringa oleifera seeds coagulate 90-95% of turbidity and color efficiently leading to an aesthetically clear supernatant & reduced about 85-90% of bacterial load reduction in samples.

Keywords: bacterial load, coagulant, turbidity, water purification

Procedia PDF Downloads 118
283 Targeting Glucocorticoid Receptor Eliminate Dormant Chemoresistant Cancer Stem Cells in Glioblastoma

Authors: Aoxue Yang, Weili Tian, Haikun Liu

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Brain tumor stem cells (BTSCs) are resistant to therapy and give rise to recurrent tumors. These rare and elusive cells are likely to disseminate during cancer progression, and some may enter dormancy, remaining viable but not increasing. The identification of dormant BTSCs is thus necessary to design effective therapies for glioblastoma (GBM) patients. Glucocorticoids (GCs) are used to treat GBM-associated edema. However, glucocorticoids participate in the physiological response to psychosocial stress, linked to poor cancer prognosis. This raises concern that glucocorticoids affect the tumor and BTSCs. Identifying markers specifically expressed by brain tumor stem cells (BTSCs) may enable specific therapies that spare their regular tissue-resident counterparts. By ribosome profiling analysis, we have identified that glycerol-3-phosphate dehydrogenase 1 (GPD1) is expressed by dormant BTSCs but not by NSCs. Through different stress-induced experiments in vitro, we found that only dexamethasone (DEXA) can significantly increase the expression of GPD1 in NSCs. Adversely, mifepristone (MIFE) which is classified as glucocorticoid receptors antagonists, could decrease GPD1 protein level and weaken the proliferation and stemness in BTSCs. Furthermore, DEXA can induce GPD1 expression in tumor-bearing mice brains and shorten animal survival, whereas MIFE has a distinct adverse effect that prolonged mice lifespan. Knocking out GR in NSC can block the upregulation of GPD1 inducing by DEXA, and we find the specific sequences on GPD1 promotor combined with GR, thus improving the efficiency of GPD1 transcription from CHIP-Seq. Moreover, GR and GPD1 are highly co-stained on GBM sections obtained from patients and mice. All these findings confirmed that GR could regulate GPD1 and loss of GPD1 Impairs Multiple Pathways Important for BTSCs Maintenance GPD1 is also a critical enzyme regulating glycolysis and lipid synthesis. We observed that DEXA and MIFE could change the metabolic profiles of BTSCs by regulating GPD1 to shift the transition of cell dormancy. Our transcriptome and lipidomics analysis demonstrated that cell cycle signaling and phosphoglycerides synthesis pathways contributed a lot to the inhibition of GPD1 caused by MIFE. In conclusion, our findings raise concern that treatment of GBM with GCs may compromise the efficacy of chemotherapy and contribute to BTSC dormancy. Inhibition of GR can dramatically reduce GPD1 and extend the survival duration of GBM-bearing mice. The molecular link between GPD1 and GR may give us an attractive therapeutic target for glioblastoma.

Keywords: cancer stem cell, dormancy, glioblastoma, glycerol-3-phosphate dehydrogenase 1, glucocorticoid receptor, dexamethasone, RNA-sequencing, phosphoglycerides

Procedia PDF Downloads 101
282 A Case Study of Remote Location Viewing, and Its Significance in Mobile Learning

Authors: James Gallagher, Phillip Benachour

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As location aware mobile technologies become ever more omnipresent, the prospect of exploiting their context awareness to enforce learning approaches thrives. Utilizing the growing acceptance of ubiquitous computing, and the steady progress both in accuracy and battery usage of pervasive devices, we present a case study of remote location viewing, how the application can be utilized to support mobile learning in situ using an existing scenario. Through the case study we introduce a new innovative application: Mobipeek based around a request/response protocol for the viewing of a remote location and explore how this can apply both as part of a teacher lead activity and informal learning situations. The system developed allows a user to select a point on a map, and send a request. Users can attach messages alongside time and distance constraints. Users within the bounds of the request can respond with an image, and accompanying message, providing context to the response. This application can be used alongside a structured learning activity such as the use of mobile phone cameras outdoors as part of an interactive lesson. An example of a learning activity would be to collect photos in the wild about plants, vegetation, and foliage as part of a geography or environmental science lesson. Another example could be to take photos of architectural buildings and monuments as part of an architecture course. These images can be uploaded then displayed back in the classroom for students to share their experiences and compare their findings with their peers. This can help to fosters students’ active participation while helping students to understand lessons in a more interesting and effective way. Mobipeek could augment the student learning experience by providing further interaction with other peers in a remote location. The activity can be part of a wider study between schools in different areas of the country enabling the sharing and interaction between more participants. Remote location viewing can be used to access images in a specific location. The choice of location will depend on the activity and lesson. For example architectural buildings of a specific period can be shared between two or more cities. The augmentation of the learning experience can be manifested in the different contextual and cultural influences as well as the sharing of images from different locations. In addition to the implementation of Mobipeek, we strive to analyse this application, and a subset of other possible and further solutions targeted towards making learning more engaging. Consideration is given to the benefits of such a system, privacy concerns, and feasibility of widespread usage. We also propose elements of “gamification”, in an attempt to further the engagement derived from such a tool and encourage usage. We conclude by identifying limitations, both from a technical, and a mobile learning perspective.

Keywords: context aware, location aware, mobile learning, remote viewing

Procedia PDF Downloads 267
281 Food Safety in Wine: Removal of Ochratoxin a in Contaminated White Wine Using Commercial Fining Agents

Authors: Antònio Inês, Davide Silva, Filipa Carvalho, Luís Filipe-Riberiro, Fernando M. Nunes, Luís Abrunhosa, Fernanda Cosme

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The presence of mycotoxins in foodstuff is a matter of concern for food safety. Mycotoxins are toxic secondary metabolites produced by certain molds, being ochratoxin A (OTA) one of the most relevant. Wines can also be contaminated with these toxicants. Several authors have demonstrated the presence of mycotoxins in wine, especially ochratoxin A. Its chemical structure is a dihydro-isocoumarin connected at the 7-carboxy group to a molecule of L-β-phenylalanine via an amide bond. As these toxicants can never be completely removed from the food chain, many countries have defined levels in food in order to attend health concerns. OTA contamination of wines might be a risk to consumer health, thus requiring treatments to achieve acceptable standards for human consumption. The maximum acceptable level of OTA in wines is 2.0 μg/kg according to the Commission regulation No. 1881/2006. Therefore, the aim of this work was to reduce OTA to safer levels using different fining agents, as well as their impact on white wine physicochemical characteristics. To evaluate their efficiency, 11 commercial fining agents (mineral, synthetic, animal and vegetable proteins) were used to get new approaches on OTA removal from white wine. Trials (including a control without addition of a fining agent) were performed in white wine artificially supplemented with OTA (10 µg/L). OTA analyses were performed after wine fining. Wine was centrifuged at 4000 rpm for 10 min and 1 mL of the supernatant was collected and added of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v). Also, the solid fractions obtained after fining, were centrifuged (4000 rpm, 15 min), the resulting supernatant discarded, and the pellet extracted with 1 mL of the above solution and 1 mL of H2O. OTA analysis was performed by HPLC with fluorescence detection. The most effective fining agent in removing OTA (80%) from white wine was a commercial formulation that contains gelatin, bentonite and activated carbon. Removals between 10-30% were obtained with potassium caseinate, yeast cell walls and pea protein. With bentonites, carboxymethylcellulose, polyvinylpolypyrrolidone and chitosan no considerable OTA removal was verified. Following, the effectiveness of seven commercial activated carbons was also evaluated and compared with the commercial formulation that contains gelatin, bentonite and activated carbon. The different activated carbons were applied at the concentration recommended by the manufacturer in order to evaluate their efficiency in reducing OTA levels. Trial and OTA analysis were performed as explained previously. The results showed that in white wine all activated carbons except one reduced 100% of OTA. The commercial formulation that contains gelatin, bentonite and activated carbon reduced only 73% of OTA concentration. These results may provide useful information for winemakers, namely for the selection of the most appropriate oenological product for OTA removal, reducing wine toxicity and simultaneously enhancing food safety and wine quality.

Keywords: wine, ota removal, food safety, fining

Procedia PDF Downloads 505
280 Experimental and Numerical Investigation of Fracture Behavior of Foamed Concrete Based on Three-Point Bending Test of Beams with Initial Notch

Authors: M. Kozłowski, M. Kadela

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Foamed concrete is known for its low self-weight and excellent thermal and acoustic properties. For many years, it has been used worldwide for insulation to foundations and roof tiles, as backfill to retaining walls, sound insulation, etc. However, in the last years it has become a promising material also for structural purposes e.g. for stabilization of weak soils. Due to favorable properties of foamed concrete, many interests and studies were involved to analyze its strength, mechanical, thermal and acoustic properties. However, these studies do not cover the investigation of fracture energy which is the core factor governing the damage and fracture mechanisms. Only limited number of publications can be found in literature. The paper presents the results of experimental investigation and numerical campaign of foamed concrete based on three-point bending test of beams with initial notch. First part of the paper presents the results of a series of static loading tests performed to investigate the fracture properties of foamed concrete of varying density. Beam specimens with dimensions of 100×100×840 mm with a central notch were tested in three-point bending. Subsequently, remaining halves of the specimens with dimensions of 100×100×420 mm were tested again as un-notched beams in the same set-up with reduced distance between supports. The tests were performed in a hydraulic displacement controlled testing machine with a load capacity of 5 kN. Apart from measuring the loading and mid-span displacement, a crack mouth opening displacement (CMOD) was monitored. Based on the load – displacement curves of notched beams the values of fracture energy and tensile stress at failure were calculated. The flexural tensile strength was obtained on un-notched beams with dimensions of 100×100×420 mm. Moreover, cube specimens 150×150×150 mm were tested in compression to determine the compressive strength. Second part of the paper deals with numerical investigation of the fracture behavior of beams with initial notch presented in the first part of the paper. Extended Finite Element Method (XFEM) was used to simulate and analyze the damage and fracture process. The influence of meshing and variation of mechanical properties on results was investigated. Numerical models simulate correctly the behavior of beams observed during three-point bending. The numerical results show that XFEM can be used to simulate different fracture toughness of foamed concrete and fracture types. Using the XFEM and computer simulation technology allow for reliable approximation of load–bearing capacity and damage mechanisms of beams made of foamed concrete, which provides some foundations for realistic structural applications.

Keywords: foamed concrete, fracture energy, three-point bending, XFEM

Procedia PDF Downloads 273
279 Mycotoxin Bioavailability in Sparus Aurata Muscle After Human Digestion and Intestinal Transport (Caco-2/HT-29 Cells) Simulation

Authors: Cheila Pereira, Sara C. Cunha, Miguel A. Faria, José O. Fernandes

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The increasing world population brings several concerns, one of which is food security and sustainability. To meet this challenge, aquaculture, the farming of aquatic animals and plants, including fish, mollusks, bivalves, and algae, has experienced sustained growth and development in recent years. Recent advances in this industry have focused on reducing its economic and environmental costs, for example, the substitution of protein sources in fish feed. Plant-based proteins are now a common approach, and while it is a greener alternative to animal-based proteins, there are some disadvantages, such as their putative content and intoxicants such as mycotoxins. These are naturally occurring plant contaminants, and their exposure in fish can cause health problems, stunted growth or even death, resulting in economic losses for the producers and health concerns for the consumers. Different works have demonstrated the presence of both AFB1 (aflatoxin B1) and ENNB1 (enniatin B1) in fish feed and their capacity to be absorbed and bioaccumulate in the fish organism after digestion, further reaching humans through fish ingestion. The aim of this work was to evaluate the bioaccessibility of both mycotoxins in samples of Sparus aurata muscle using a static digestion model based on the INFOGEST protocol. The samples were subjected to different cooking procedures – raw, grilled and fried – and different seasonings – none, thyme and ginger – in order to evaluate their potential reduction effect on mycotoxins bioaccessibility, followed by the evaluation of the intestinal transport of both compounds with an in vitro cell model composed of Caco-2/HT-29 co-culture monolayers, simulating the human intestinal epithelium. The bioaccessible fractions obtained in the digestion studies were used in the transport studies for a more realistic approach to bioavailability evaluation. Results demonstrated the effect of the use of different cooking procedures and seasoning on the toxin's bioavailability. Sparus aurata was chosen in this study for its large production in aquaculture and high consumption in Europe. Also, with the continued evolution of fish farming practices and more common usage of novel feed ingredients based on plants, there is a growing concern about less studied contaminants in aquaculture and their consequences for human health. In pair with greener advances in this industry, there is a convergence towards alternative research methods, such as in vitro applications. In the case of bioavailability studies, both in vitro digestion protocols and intestinal transport assessment are excellent alternatives to in vivo studies. These methods provide fast, reliable and comparable results without ethical restraints.

Keywords: AFB1, aquaculture, bioaccessibility, ENNB1, intestinal transport.

Procedia PDF Downloads 30
278 Landslide Susceptibility Analysis in the St. Lawrence Lowlands Using High Resolution Data and Failure Plane Analysis

Authors: Kevin Potoczny, Katsuichiro Goda

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The St. Lawrence lowlands extend from Ottawa to Quebec City and are known for large deposits of sensitive Leda clay. Leda clay deposits are responsible for many large landslides, such as the 1993 Lemieux and 2010 St. Jude (4 fatalities) landslides. Due to the large extent and sensitivity of Leda clay, regional hazard analysis for landslides is an important tool in risk management. A 2018 regional study by Farzam et al. on the susceptibility of Leda clay slopes to landslide hazard uses 1 arc second topographical data. A qualitative method known as Hazus is used to estimate susceptibility by checking for various criteria in a location and determine a susceptibility rating on a scale of 0 (no susceptibility) to 10 (very high susceptibility). These criteria are slope angle, geological group, soil wetness, and distance from waterbodies. Given the flat nature of St. Lawrence lowlands, the current assessment fails to capture local slopes, such as the St. Jude site. Additionally, the data did not allow one to analyze failure planes accurately. This study majorly improves the analysis performed by Farzam et al. in two aspects. First, regional assessment with high resolution data allows for identification of local locations that may have been previously identified as low susceptibility. This then provides the opportunity to conduct a more refined analysis on the failure plane of the slope. Slopes derived from 1 arc second data are relatively gentle (0-10 degrees) across the region; however, the 1- and 2-meter resolution 2022 HRDEM provided by NRCAN shows that short, steep slopes are present. At a regional level, 1 arc second data can underestimate the susceptibility of short, steep slopes, which can be dangerous as Leda clay landslides behave retrogressively and travel upwards into flatter terrain. At the location of the St. Jude landslide, slope differences are significant. 1 arc second data shows a maximum slope of 12.80 degrees and a mean slope of 4.72 degrees, while the HRDEM data shows a maximum slope of 56.67 degrees and a mean slope of 10.72 degrees. This equates to a difference of three susceptibility levels when the soil is dry and one susceptibility level when wet. The use of GIS software is used to create a regional susceptibility map across the St. Lawrence lowlands at 1- and 2-meter resolutions. Failure planes are necessary to differentiate between small and large landslides, which have so far been ignored in regional analysis. Leda clay failures can only retrogress as far as their failure planes, so the regional analysis must be able to transition smoothly into a more robust local analysis. It is expected that slopes within the region, once previously assessed at low susceptibility scores, contain local areas of high susceptibility. The goal is to create opportunities for local failure plane analysis to be undertaken, which has not been possible before. Due to the low resolution of previous regional analyses, any slope near a waterbody could be considered hazardous. However, high-resolution regional analysis would allow for more precise determination of hazard sites.

Keywords: hazus, high-resolution DEM, leda clay, regional analysis, susceptibility

Procedia PDF Downloads 49
277 Influence of Dietary Boron on Gut Absorption of Nutrients, Blood Metabolites and Tissue Pathology

Authors: T. Vijay Bhasker, N. K. S Gowda, P. Krishnamoorthy, D. T. Pal, A. K. Pattanaik, A. K. Verma

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Boron (B) is a newer trace element and its biological importance and dietary essentiality is unclear in animals. The available literature suggests its putative role in bone mineralization, antioxidant status and steroid hormone synthesis. A feeding trial was conducted in Wister strain (Rattus norvegicus) albino rats for duration of 90 days. A total of 84 healthy weaned (3-4 weeks) experimental rats were randomly divided into 7 dietary groups (4 replicates of three each) viz., A (Basal diet/ Control), B (Basal diet + 5 ppm B), C (Basal diet + 10 ppm B), D (Basal diet + 20 ppm B), E (Basal diet + 40 ppm B), F (Basal diet-Ca 50%), G (Basal diet-Ca 50% + 40 ppm B). Dietary level of calcium (Ca) was maintained at two levels, 100% and 50% of requirement. Sodium borate was used as source of boron along with other ingredients of basal diet while preparing the pelletized diets. All the rats were kept in proper ventilated laboratory animal house maintained at temperature (23±2º C) and humidity (50 to 70%). At the end of experiment digestibility trial was conducted for 5 days to estimate nutrient digestibility and gut absorption of minerals. Eight rats from each group were sacrificed to collect the vital organs (liver, kidney and spleen) to study histopathology. Blood sample was drawn by heart puncture to determine biochemical profile. The average daily feed intake (g/rat/day), water intake (ml/rat/day) and body weight gain (g/rat/day) were similar among the dietary groups. The digestibility (%) of organic matter and crude fat were significantly improved (P < 0.05) was by B supplementation. The gut absorption (%) Ca was significantly increased (P < 0.01) in B supplemented groups compared to control. However, digestibility of dry matter and crude protein, gut absorption of magnesium and phosphorus showed a non-significant increasing trend with B supplementation. The gut absorption (%) of B (P < 0.01) was significantly lowered (P<0.05) in supplemented groups compared to un-supplemented ones. The serum level of triglycerides (mg/dL), HDL-cholesterol (mg/dL) and alanine transaminase (IU/L) were significantly lowered (P < 0.05) in B supplemented groups. While serum level of glucose (mg/dL) and alkaline phosphatase (KA units) showed a non-significant decreasing trend with B supplementation. However the serum levels of total cholesterol (mg/dL) and aspartate transaminase (IU/L) were similar among dietary groups. The histology sections of kidney and spleen revealed no significant changes among the dietary groups and were observed to be normal in anatomical architecture. However, the liver histology revealed cell degenerative changes with vacuolar degeneration and nuclear condensation in Ca deficient groups. But the comparative degenerative changes were mild in 40 ppm B supplemented Ca deficient group. In conclusion, dietary supplementation of graded levels of boron in rats had a positive effect on metabolism and health by improving nutrient digestibility and gut absorption of Ca. This indicates the beneficial role of dietary boron supplementation.

Keywords: boron, calcium, nutrient utilization, histopathology

Procedia PDF Downloads 299
276 Multi-Criteria Selection and Improvement of Effective Design for Generating Power from Sea Waves

Authors: Khaled M. Khader, Mamdouh I. Elimy, Omayma A. Nada

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Sustainable development is the nominal goal of most countries at present. In general, fossil fuels are the development mainstay of most world countries. Regrettably, the fossil fuel consumption rate is very high, and the world is facing the problem of conventional fuels depletion soon. In addition, there are many problems of environmental pollution resulting from the emission of harmful gases and vapors during fuel burning. Thus, clean, renewable energy became the main concern of most countries for filling the gap between available energy resources and their growing needs. There are many renewable energy sources such as wind, solar and wave energy. Energy can be obtained from the motion of sea waves almost all the time. However, power generation from solar or wind energy is highly restricted to sunny periods or the availability of suitable wind speeds. Moreover, energy produced from sea wave motion is one of the cheapest types of clean energy. In addition, renewable energy usage of sea waves guarantees safe environmental conditions. Cheap electricity can be generated from wave energy using different systems such as oscillating bodies' system, pendulum gate system, ocean wave dragon system and oscillating water column device. In this paper, a multi-criteria model has been developed using Analytic Hierarchy Process (AHP) to support the decision of selecting the most effective system for generating power from sea waves. This paper provides a widespread overview of the different design alternatives for sea wave energy converter systems. The considered design alternatives have been evaluated using the developed AHP model. The multi-criteria assessment reveals that the off-shore Oscillating Water Column (OWC) system is the most appropriate system for generating power from sea waves. The OWC system consists of a suitable hollow chamber at the shore which is completely closed except at its base which has an open area for gathering moving sea waves. Sea wave's motion pushes the air up and down passing through a suitable well turbine for generating power. Improving the power generation capability of the OWC system is one of the main objectives of this research. After investigating the effect of some design modifications, it has been concluded that selecting the appropriate settings of some effective design parameters such as the number of layers of Wells turbine fans and the intermediate distance between the fans can result in significant improvements. Moreover, simple dynamic analysis of the Wells turbine is introduced. Furthermore, this paper strives for comparing the theoretical and experimental results of the built experimental prototype.

Keywords: renewable energy, oscillating water column, multi-criteria selection, Wells turbine

Procedia PDF Downloads 137
275 Influence of Iron Content in Carbon Nanotubes on the Intensity of Hyperthermia in the Cancer Treatment

Authors: S. Wiak, L. Szymanski, Z. Kolacinski, G. Raniszewski, L. Pietrzak, Z. Staniszewska

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The term ‘cancer’ is given to a collection of related diseases that may affect any part of the human body. It is a pathological behaviour of cells with the potential to undergo abnormal breakdown in the processes that control cell proliferation, differentiation, and death of particular cells. Although cancer is commonly considered as modern disease, there are beliefs that drastically growing number of new cases can be linked to the extensively prolonged life expectancy and enhanced techniques for cancer diagnosis. Magnetic hyperthermia therapy is a novel approach to cancer treatment, which may greatly contribute to higher efficiency of the therapy. Employing carbon nanotubes as nanocarriers for magnetic particles, it is possible to decrease toxicity and invasiveness of the treatment by surface functionalisation. Despite appearing in recent years, magnetic particle hyperthermia has already become of the highest interest in the scientific and medical environment. The reason why hyperthermia therapy brings so much hope for future treatment of cancer lays in the effect that it produces in malignant cells. Subjecting them to thermal shock results in activation of numerous degradation processes inside and outside the cell. The heating process initiates mechanisms of DNA destruction, protein denaturation and induction of cell apoptosis, which may lead to tumour shrinkage, and in some cases, it may even cause complete disappearance of cancer. The factors which have the major impact on the final efficiency of the treatment include temperatures generated inside the tissues, time of exposure to the heating process, and the character of an individual cancer cell type. The vast majority of cancer cells is characterised by lower pH, persistent hypoxia and lack of nutrients, which can be associated to abnormal microvasculature. Since in healthy tissues we cannot observe presence of these conditions, they should not be seriously affected by elevation of the temperature. The aim of this work is to investigate the influence of iron content in iron filled Carbon Nanotubes on the desired nanoparticles for cancer therapy. In the article, the development and demonstration of the method and the model device for hyperthermic selective destruction of cancer cells are presented. This method was based on the synthesis and functionalization of carbon nanotubes serving as ferromagnetic material nanocontainers. The methodology of the production carbon- ferromagnetic nanocontainers (FNCs) includes the synthesis of carbon nanotubes, chemical, and physical characterization, increasing the content of a ferromagnetic material and biochemical functionalization involving the attachment of the key addresses. The ferromagnetic nanocontainers were synthesised in CVD and microwave plasma system. The research work has been financed from the budget of science as a research project No. PBS2/A5/31/2013.

Keywords: hyperthermia, carbon nanotubes, cancer colon cells, radio frequency field

Procedia PDF Downloads 104
274 Fodder Production and Livestock Rearing in Relation to Climate Change and Possible Adaptation Measures in Manaslu Conservation Area, Nepal

Authors: Bhojan Dhakal, Naba Raj Devkota, Chet Raj Upreti, Maheshwar Sapkota

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A study was conducted to find out the production potential, nutrient composition, and the variability of the most commonly available fodder trees along with the varying altitude to help optimize the dry matter requirement during winter lean period. The study was carried out from March to June, 2012 in Lho and Prok Village Development Committee of Manaslu Conservation Area (MCA), located in Gorkha district of Nepal. The other objective of the research was to learn the impact of climate change on livestock production linking it with feed availability. The study was conducted in two parts: social and biological. Accordingly, a households (HHs) survey was conducted to collect primary data from 70 HHs, focusing on the perception of respondents on impacts of climatic variability on the feeding management. The next part consisted of understanding yield potential and nutrient composition of the four most commonly available fodder trees (M. azedirach, M. alba, F. roxburghii, F. nemoralis), within two altitudes range: (1500-2000 masl and 2000-2500 masl) by using a RCB design in 2*4 factorial combination of treatments, each replicated four times. Results revealed that majority of the farmers perceived the change in climatic phenomenon more severely within the past five years. Farmers were using different adaptation technologies such as collection of forage from jungle, reducing unproductive animals, fodder trees utilization, and crop by product feeding at feed scarcity period. Ranking of the different fodder trees on the basis of indigenous knowledge and experiences revealed that F. roxburghii was the best-preferred fodder tree species (index value 0.72) in terms overall preferability whereas M. azedirach had highest growth and productivity (index value 0.77), F. roxburghii had highest adoptability (index value 0.69) and palatability (index value 0.69) as well. Similarly, fresh yield and dry matter yield of the each fodder trees was significant (P < 0.01) between the altitude and within species. Fodder trees yield analysis revealed that the highest dry matter (DM) yield (28 kg/tree) was obtained for F. roxburghii but that remained statistically similar (P > 0.05) to the other treatment. On the other hand, most of the parameters: ether extract (EE), acid detergent lignin (ADL), acid detergent fibre (ADF), cell wall digestibility (CWD), relative digestibility (RD), digestible nutrient (TDN), and Calcium (Ca) among the treatments were highly significant (P < 0.01). This indicates the scope of introducing productive and nutritive fodder trees species even at the high altitude to help reduce fodder scarcity problem during winter. The finding also revealed the scope of promoting all available local fodder trees species as crude protein content of these species were similar.

Keywords: fodder trees, yield potential, climate change, nutrient composition

Procedia PDF Downloads 278
273 Global Experiences in Dealing with Biological Epidemics with an Emphasis on COVID-19 Disease: Approaches and Strategies

Authors: Marziye Hadian, Alireza Jabbari

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Background: The World Health Organization has identified COVID-19 as a public health emergency and is urging governments to stop the virus transmission by adopting appropriate policies. In this regard, authorities have taken different approaches to cut the chain or controlling the spread of the disease. Now, the questions we are facing include what these approaches are? What tools should be used to implement each preventive protocol? In addition, what is the impact of each approach? Objective: The aim of this study was to determine the approaches to biological epidemics and related prevention tools with an emphasis on COVID-19 disease. Data sources: Databases including ISI web of science, PubMed, Scopus, Science Direct, Ovid, and ProQuest were employed for data extraction. Furthermore, authentic sources such as the WHO website, the published reports of relevant countries, as well as the Worldometer website were evaluated for gray studies. The time-frame of the study was from 1 December 2019 to 30 May 2020. Methods: The present study was a systematic study of publications related to the prevention strategies for the COVID-19 disease. The study was carried out based on the PRISMA guidelines and CASP for articles and AACODS for grey literature. Results: The study findings showed that in order to confront the COVID-19 epidemic, in general, there are three approaches of "mitigation", "active control" and "suppression" and four strategies of "quarantine", "isolation", "social distance" and "lockdown" in both individual and social dimensions to deal with epidemics. Selection and implementation of each approach requires specific strategies and has different effects when it comes to controlling and inhibiting the disease. Key finding: One possible approach to control the disease is to change individual behavior and lifestyle. In addition to prevention strategies, use of masks, observance of personal hygiene principles such as regular hand washing and non-contact of contaminated hands with the face, as well as an observance of public health principles such as sneezing and coughing etiquettes, safe extermination of personal protective equipment, must be strictly observed. Have not been included in the category of prevention tools. However, it has a great impact on controlling the epidemic, especially the new coronavirus epidemic. Conclusion: Although the use of different approaches to control and inhibit biological epidemics depends on numerous variables, however, despite these requirements, global experience suggests that some of these approaches are ineffective. The use of previous experiences in the world, along with the current experiences of countries, can be very helpful in choosing the accurate approach for each country in accordance with the characteristics of that country and lead to the reduction of possible costs at the national and international levels.

Keywords: novel corona virus, COVID-19, approaches, prevention tools, prevention strategies

Procedia PDF Downloads 105
272 Stable Time Reversed Integration of the Navier-Stokes Equation Using an Adjoint Gradient Method

Authors: Jurriaan Gillissen

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This work is concerned with stabilizing the numerical integration of the Navier-Stokes equation (NSE), backwards in time. Applications involve the detection of sources of, e.g., sound, heat, and pollutants. Stable reverse numerical integration of parabolic differential equations is also relevant for image de-blurring. While the literature addresses the reverse integration problem of the advection-diffusion equation, the problem of numerical reverse integration of the NSE has, to our knowledge, not yet been addressed. Owing to the presence of viscosity, the NSE is irreversible, i.e., when going backwards in time, the fluid behaves, as if it had a negative viscosity. As an effect, perturbations from the perfect solution, due to round off errors or discretization errors, grow exponentially in time, and reverse integration of the NSE is inherently unstable, regardless of using an implicit time integration scheme. Consequently, some sort of filtering is required, in order to achieve a stable, numerical, reversed integration. The challenge is to find a filter with a minimal adverse affect on the accuracy of the reversed integration. In the present work, we explore an adjoint gradient method (AGM) to achieve this goal, and we apply this technique to two-dimensional (2D), decaying turbulence. The AGM solves for the initial velocity field u0 at t = 0, that, when integrated forward in time, produces a final velocity field u1 at t = 1, that is as close as is feasibly possible to some specified target field v1. The initial field u0 defines a minimum of a cost-functional J, that measures the distance between u1 and v1. In the minimization procedure, the u0 is updated iteratively along the gradient of J w.r.t. u0, where the gradient is obtained by transporting J backwards in time from t = 1 to t = 0, using the adjoint NSE. The AGM thus effectively replaces the backward integration by multiple forward and backward adjoint integrations. Since the viscosity is negative in the adjoint NSE, each step of the AGM is numerically stable. Nevertheless, when applied to turbulence, the AGM develops instabilities, which limit the backward integration to small times. This is due to the exponential divergence of phase space trajectories in turbulent flow, which produces a multitude of local minima in J, when the integration time is large. As an effect, the AGM may select unphysical, noisy initial conditions. In order to improve this situation, we propose two remedies. First, we replace the integration by a sequence of smaller integrations, i.e., we divide the integration time into segments, where in each segment the target field v1 is taken as the initial field u0 from the previous segment. Second, we add an additional term (regularizer) to J, which is proportional to a high-order Laplacian of u0, and which dampens the gradients of u0. We show that suitable values for the segment size and for the regularizer, allow a stable reverse integration of 2D decaying turbulence, with accurate results for more then O(10) turbulent, integral time scales.

Keywords: time reversed integration, parabolic differential equations, adjoint gradient method, two dimensional turbulence

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271 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

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

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

Procedia PDF Downloads 245
270 A Numerical Studies for Improving the Performance of Vertical Axis Wind Turbine by a Wind Power Tower

Authors: Soo-Yong Cho, Chong-Hyun Cho, Chae-Whan Rim, Sang-Kyu Choi, Jin-Gyun Kim, Ju-Seok Nam

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Recently, vertical axis wind turbines (VAWT) have been widely used to produce electricity even in urban. They have several merits such as low sound noise, easy installation of the generator and simple structure without yaw-control mechanism and so on. However, their blades are operated under the influence of the trailing vortices generated by the preceding blades. This phenomenon deteriorates its output power and makes difficulty predicting correctly its performance. In order to improve the performance of VAWT, wind power towers can be applied. Usually, the wind power tower can be constructed as a multi-story building to increase the frontal area of the wind stream. Hence, multiple sets of the VAWT can be installed within the wind power tower, and they can be operated at high elevation. Many different types of wind power tower can be used in the field. In this study, a wind power tower with circular column shape was applied, and the VAWT was installed at the center of the wind power tower. Seven guide walls were used as a strut between the floors of the wind power tower. These guide walls were utilized not only to increase the wind velocity within the wind power tower but also to adjust the wind direction for making a better working condition on the VAWT. Hence, some important design variables, such as the distance between the wind turbine and the guide wall, the outer diameter of the wind power tower, the direction of the guide wall against the wind direction, should be considered to enhance the output power on the VAWT. A numerical analysis was conducted to find the optimum dimension on design variables by using the computational fluid dynamics (CFD) among many prediction methods. The CFD could be an accurate prediction method compared with the stream-tube methods. In order to obtain the accurate results in the CFD, it needs the transient analysis and the full three-dimensional (3-D) computation. However, this full 3-D CFD could be hard to be a practical tool because it requires huge computation time. Therefore, the reduced computational domain is applied as a practical method. In this study, the computations were conducted in the reduced computational domain and they were compared with the experimental results in the literature. It was examined the mechanism of the difference between the experimental results and the computational results. The computed results showed this computational method could be an effective method in the design methodology using the optimization algorithm. After validation of the numerical method, the CFD on the wind power tower was conducted with the important design variables affecting the performance of VAWT. The results showed that the output power of the VAWT obtained using the wind power tower was increased compared to them obtained without the wind power tower. In addition, they showed that the increased output power on the wind turbine depended greatly on the dimension of the guide wall.

Keywords: CFD, performance, VAWT, wind power tower

Procedia PDF Downloads 360
269 Genetic Diversity of Cord Blood of the National Center of Blood Transfusion, Mexico (NCBT)

Authors: J. Manuel Bello-López, Julieta Rojo-Medina

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Introduction: The transplant of Umbilical Cord Blood Units (UCBU) are a therapeutic possibility for patients with oncohaematological disorders, especially in children. In Mexico, 48.5% of oncological diseases in children 1-4 years old are leukemias; whereas in patients 5-14 and 15-24 years old, lymphomas and leukemias represent the second and third cause of death in these groups respectively. Therefore it is necessary to have more registries of UCBU in order to ensure genetic diversity in the country; the above because the search for appropriate a UCBU is increasingly difficult for patients of mixed ethnicity. Objective: To estimate the genetic diversity (polymorphisms) of Human Leucocyte Antigen (HLA) Class I (A, B) and Class II (DRB1) in UCBU cryopreserved for transplant at Cord Blood Bank of the NCBT. Material and Methods: HLA typing of 533 UCBU for transplant was performed from 2003-2012 at the Histocompatibility Laboratory from the Research Department (evaluated by Los Angeles Ca. Immunogenetics Center) of the NCBT. Class I HLA-A, HLA-B and Class II HLA-DRB1 typing was performed using medium resolution Sequence-Specific Primer (SSP). In cases of an ambiguity detected by SSP; Sequence-Specific Oligonucleotide (SSO) method was carried out. A strict analysis of populations genetic parameters were done in 5 representative UCBU populations. Results: 46.5% of UCBU were collected from Mexico City, State of Mexico (30.95%), Puebla (8.06%), Morelos (6.37%) and Veracruz (3.37%). The remaining UCBU (4.75%) are represented by other states. The identified genotypes correspond to Amerindian origins (HLA-A*02, 31; HLA-B*39, 15, 48), Caucasian (HLA-A*02, 68, 01, 30, 31; HLA-B*35, 15, 40, 44, 07 y HLA-DRB1*04, 08, 07, 15, 03, 14), Oriental (HLA-A*02, 30, 01, 31; HLA-B* 35, 39, 15, 40, 44, 07,48 y HLA-DRB1*04, 07,15, 03) and African (HLA-A*30 y HLA-DRB1*03). The genetic distances obtained by Cavalli-Sforza analysis of the five states showed significant genetic differences by comparing genetic frequencies. The shortest genetic distance exists between Mexico City and the state of Puebla (0.0039) and the largest between Veracruz and Morelos (0.0084). In order to identify significant differences between this states, the ANOVA test was performed. This demonstrates that UCBU is significantly different according to their origin (P <0.05). This is shown by the divergence between arms at the Dendogram of Neighbor-Joining. Conclusions: The NCBT provides UCBU in patients with oncohaematological disorders in all the country. There is a group of patients for which not compatible UCBU can be find due to the mixed ethnic origin. For example, the population of northern Mexico is mostly Caucasian. Most of the NCBT donors are of various ethnic origins, predominantly Amerindians and Caucasians; although some ethnic minorities like Oriental, African and pure Indian ethnics are not represented. The NCBT is, therefore, establishing agreements with different states of Mexico to promote the altruistic donation of Umbilical Cord Blood in order to enrich the genetic diversity in its files.

Keywords: cord blood, genetic diversity, human leucocyte antigen, transplant

Procedia PDF Downloads 361
268 The Hidden Mechanism beyond Ginger (Zingiber officinale Rosc.) Potent in vivo and in vitro Anti-Inflammatory Activity

Authors: Shahira M. Ezzat, Marwa I. Ezzat, Mona M. Okba, Esther T. Menze, Ashraf B. Abdel-Naim, Shahnas O. Mohamed

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Background: In order to decrease the burden of the high cost of synthetic drugs, it is important to focus on phytopharmaceuticals. The aim of our study was to search for the mechanism of ginger (Zingiber officinale Roscoe) anti-inflammatory potential and to correlate it to its biophytochemicals. Methods: Various extracts viz. water, 50%, 70%, 80%, and 90% ethanol were prepared from ginger rhizomes. Fractionation of the aqueous extract (AE) was accomplished using Diaion HP-20. In vitro anti-inflammatory activity of the different extracts and isolated compounds was evaluated by protein denaturation inhibition, membrane stabilization, protease inhibition, and anti-lipoxygenase assays. In vivo anti-inflammatory activity of AE was estimated by assessment of rat paw oedema after carrageenan injection. Prostaglandin E2 (PGE2), certain inflammation markers (TNF-α, IL-6, IL-1α, IL-1β, INFr, MCP-1MIP, RANTES, and Nox) levels and MPO activity in the paw edema exudates were measured. Total antioxidant capacity (TAC) was also determined. Histopathological alterations of paw tissues were scored. Results: All the tested extracts showed significant (p < 0.1) anti-inflammatory activities. The highest percentage of heat induced albumin denaturation (66%) was exhibited by the 50% ethanol (250 μg/ml). The 70 and 90% ethanol extracts (500 μg/ml) were more potent as membrane stabilizers (34.5 and 37%, respectively) than diclofenac (33%). The 80 and 90% ethanol extracts (500 μg/ml) showed maximum protease inhibition (56%). The strongest anti-lipoxygenase activity was observed for the AE. It showed more significant lipoxygenase inhibition activity than that of diclofenac (58% and 52%, respectively) at the same concentration (125 μg/ml). Fractionation of AE yielded four main fractions (Fr I-IV) which showed significant in vitro anti-inflammatory. Purification of Fr-III and IV led to the isolation of 6-poradol (G1), 6-shogaol (G2); methyl 6- gingerol (G3), 5-gingerol (G4), 6-gingerol (G5), 8-gingerol (G6), 10-gingerol (G7), and 1-dehydro-6-gingerol (G8). G2 (62.5 ug/ml), G1 (250 ug/ml), and G8 (250 ug/ml) exhibited potent anti-inflammatory activity in all studied assays, while G4 and G5 exhibited moderate activity. In vivo administration of AE ameliorated rat paw oedema in a dose-dependent manner. AE (at 200 mg/kg) showed significant reduction (60%) of PGE2 production. The AE at different doses (at 25-200 mg/kg) showed significant reduction in inflammatory markers except for IL-1α. AE (at 25 mg/kg) is superior to indomethacin in reduction of IL-1β. Treatment of animals with the AE (100, 200 mg/kg) or indomethacin (10 mg/kg) showed significant reduction in TNF-α, IL-6, MCP-1, and RANTES levels, and MPO activity by about (31, 57 and 32% ) (65, 60 and 57%) (27, 41 and 28%) (23, 32 and 23%) (66, 67 and 67%) respectively. AE at 100 and 200 mg/kg was equipotent to indomethacin in reduction of NOₓ level and in increasing the TAC. Histopathological examination revealed very few inflammatory cells infiltration and oedema after administration of AE (200 mg/kg) prior to carrageenan. Conclusion: Ginger anti-inflammatory activity is mediated by inhibiting macrophage and neutrophils activation as well as negatively affecting monocyte and leukocyte migration. Moreover, it produced dose-dependent decrease in pro-inflammatory cytokines and chemokines and replenished the total antioxidant capacity. We strongly recommend future investigations of ginger in the potential signal transduction pathways.

Keywords: anti-lipoxygenase activity, inflammatory markers, 1-dehydro-6-gingerol, 6-shogaol

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267 The Potential of On-Demand Shuttle Services to Reduce Private Car Use

Authors: B. Mack, K. Tampe-Mai, E. Diesch

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Findings of an ongoing discrete choice study of future transport mode choice will be presented. Many urban centers face the triple challenge of having to cope with ever increasing traffic congestion, environmental pollution, and greenhouse gas emission brought about by private car use. In principle, private car use may be diminished by extending public transport systems like bus lines, trams, tubes, and trains. However, there are limits to increasing the (perceived) spatial and temporal flexibility and reducing peak-time crowding of classical public transport systems. An emerging new type of system, publicly or privately operated on-demand shuttle bus services, seem suitable to ameliorate the situation. A fleet of on-demand shuttle busses operates without fixed stops and schedules. It may be deployed efficiently in that each bus picks up passengers whose itineraries may be combined into an optimized route. Crowding may be minimized by limiting the number of seats and the inter-seat distance for each bus. The study is conducted as a discrete choice experiment. The choice between private car, public transport, and shuttle service is registered as a function of several push and pull factors (financial costs, travel time, walking distances, mobility tax/congestion charge, and waiting time/parking space search time). After the completion of the discrete choice items, the study participant is asked to rate the three modes of transport with regard to the pull factors of comfort, safety, privacy, and opportunity to engage in activities like reading or surfing the internet. These ratings are entered as additional predictors into the discrete choice experiment regression model. The study is conducted in the region of Stuttgart in southern Germany. N=1000 participants are being recruited. Participants are between 18 and 69 years of age, hold a driver’s license, and live in the city or the surrounding region of Stuttgart. In the discrete choice experiment, participants are asked to assume they lived within the Stuttgart region, but outside of the city, and were planning the journey from their apartment to their place of work, training, or education during the peak traffic time in the morning. Then, for each item of the discrete choice experiment, they are asked to choose between the transport modes of private car, public transport, and on-demand shuttle in the light of particular values of the push and pull factors studied. The study will provide valuable information on the potential of switching from private car use to the use of on-demand shuttles, but also on the less desirable potential of switching from public transport to on-demand shuttle services. Furthermore, information will be provided on the modulation of these switching potentials by pull and push factors.

Keywords: determinants of travel mode choice, on-demand shuttle services, private car use, public transport

Procedia PDF Downloads 153
266 MTT Assay-Guided Isolation of a Cytotoxic Lead from Hedyotis umbellata and Its Mechanism of Action against Non-Small Cell Lung Cancer A549 Cells

Authors: Kirti Hira, A. Sajeli Begum, S. Mahibalan, Poorna Chandra Rao

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Introduction: Cancer is one of the leading causes of death worldwide. Although existing therapy effectively kills cancer cells, they do affect normal growing cells leading to many undesirable side effects. Hence there is need to develop effective as well as safe drug molecules to combat cancer, which is possible through phyto-research. The currently available plant-derived blockbuster drugs are the example for this. In view of this, an investigation was done to identify cytotoxic lead molecules from Hedyotis umbellata (Family Rubiaceae), a widely distributed weed in India. Materials and Methods: The methanolic extract of the whole plant of H. umbellata (MHU), prepared through Soxhlet extraction method was further fractionated with diethyl ether and n-butanol, successively. MHU, ether fraction (EMHU) and butanol fraction (BMHU) were lyophilized and were tested for the cytotoxic effect using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay against non-small cell lung cancer (NSCLC) A549 cell lines. The potentially active EMHU was subjected to chromatographic purification using normal-phase silica columns, in order to isolate the responsible bioactive compounds. The isolated pure compounds were tested for their cytotoxic effect by MTT assay against A549 cells. Compound-3, which was found to be most active, was characterized using IR, 1H- and 13C-NMR and MS analysis. The study was further extended to decipher the mechanism of action of cytotoxicity of compound-3 against A549 cells through various in vitro cellular models. Cell cycle analysis was done using flow cytometry following PI (Propidium Iodide) staining. Protein analysis was done using Western blot technique. Results: Among MHU, EMHU, and BMHU, the non-polar fraction EMHU demonstrated a significant dose-dependent cytotoxic effect with IC50 of 67.7μg/ml. Chromatography of EMHU yielded seven compounds. MTT assay of isolated compounds explored compound-3 as potentially active one, which inhibited the growth of A549 cells with IC50value of 14.2μM. Further, compound-3 was identified as cedrelopsin, a coumarin derivative having molecular weight of 260. Results of in vitro mechanistic studies explained that cedrelopsin induced cell cycle arrest at G2/M phase and down-regulated the expression of G2/M regulatory proteins such as cyclin B1, cdc2, and cdc25C, dose dependently. This is the first report that explores the cytotoxic mechanism of cedrelopsin. Conclusion: Thus a potential small lead molecule, cedrelopsin isolated from H. umbellata, showing antiproliferative effect mediated by G2/M arrest in A549 cells was discovered. The effect of cedrelopsin against other cancer cell lines followed by in vivo studies can be performed in future to develop a new drug candidate.

Keywords: A549, cedrelopsin, G2/M phase, Hedyotis umbellata

Procedia PDF Downloads 152
265 Enhancing Chinese Foreign Language Teachers’ Intercultural Competence: An Action Research Study

Authors: Wei Hing Rosenkvist

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In the past few decades, concerns and demands of promoting student intercultural communicative competence in foreign language education have been increasing along with the rapid growth of information technologies and globalization in the 21st century. In Sweden, related concepts such as internationalization, global citizenship, multiculturalism, and intercultural communication, are also keywords that would be found in the written learning objectives of foreign language education at all levels. Being one of the leading higher institutes in distance education in Europe, Dalarna University clearly states that after completion of the teacher education program, students shall understand the needs for integrating internationalization, intercultural and global perspective in teaching and learning in Swedish schools and implement their studies to promote education in an international and global context. Even though many teachers and educators agree with the institutes’ mission and vision about the importance of internationalization and the need to increase student understanding of intercultural and global perspectives, they might find this objective unattainable and restricted due to the nature of the subject and their knowledge of intercultural competence. When conducting a comprehensive Chinese language course for the students who are going to become Chinese foreign language teachers, the researcher found that all the learning objectives are linguistic oriented while grammatical components dominate the entire course. Apparently, there is a gap between the learning objectives of the course and the DU’s mission of fostering an international learner with intercultural and globalized perspectives. How to include this macro-learning objective in a foreign language course is a great challenge to the educator. Although scholars from different academic domains have provided different theoretical frameworks and approaches for developing student intercultural competence, research that focuses on the didactic perspectives of developing student intercultural competence in teaching Chinese as a foreign language education (CFL) is limited, and practical examples are rare. This challenge has motivated the researcher to conduct an action research study that aims at integrating DU’s macro-learning objective in a current CFL course through different didactic practices to develop the student's intercultural competence. This research study aims to, firstly, illustrate the cross-cultural knowledge integrated into the present Chinese language course for developing intercultural competence. Secondly, it investigates different didactic means that can be utilized to deliver cross-cultural knowledge to student teachers in the present course without generating dramatic disturbance of the syllabus. Thirdly, it examines the effectiveness of these didactic means in enhancing student-teacher intercultural competence regarding the need for integrating and implementing internationalization, intercultural and global perspectives in teaching and learning in Swedish schools. Last but not least, it intends to serve as a practical example for developing the student teachers’ intercultural competence in foreign language education in DU and fill in the research gap of this academic domain worldwide.

Keywords: action research, intercultural competence, Chinese as a foreign language education, teacher education

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264 The Temporal Implications of Spatial Prospects

Authors: Zhuo Job Chen, Kevin Nute

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The work reported examines potential linkages between spatial and temporal prospects, and more specifically, between variations in the spatial depth and foreground obstruction of window views, and observers’ sense of connection to the future. It was found that external views from indoor spaces were strongly associated with a sense of the future, that partially obstructing such a view with foreground objects significantly reduced its association with the future, and replacing it with a pictorial representation of the same scene (with no real actual depth) removed most of its temporal association. A lesser change in the spatial depth of the view, however, had no apparent effect on association with the future. While the role of spatial depth has still to be confirmed, the results suggest that spatial prospects directly affect temporal ones. The word “prospect” typifies the overlapping of the spatial and temporal in most human languages. It originated in classical times as a purely spatial term, but in the 16th century took on the additional temporal implication of an imagined view ahead, of the future. The psychological notion of prospection, then, has its distant origins in a spatial analogue. While it is not yet proven that space directly structures our processing of time at a physiological level, it is generally agreed that it commonly does so conceptually. The mental representation of possible futures has been a central part of human survival as a species (Boyer, 2008; Suddendorf & Corballis, 2007). A sense of the future seems critical not only practically, but also psychologically. It has been suggested, for example, that lack of a positive image of the future may be an important contributing cause of depression (Beck, 1974; Seligman, 2016). Most people in the developed world now spend more than 90% of their lives indoors. So any direct link between external views and temporal prospects could have important implications for both human well-being and building design. We found that the ability to see what lies in front of us spatially was strongly associated with a sense of what lies ahead temporally. Partial obstruction of a view was found to significantly reduce that sense connection to the future. Replacing a view with a flat pictorial representation of the same scene removed almost all of its connection with the future, but changing the spatial depth of a real view appeared to have no significant effect. While foreground obstructions were found to reduce subjects’ sense of connection to the future, they increased their sense of refuge and security. Consistent with Prospect and Refuge theory, an ideal environment, then, would seem to be one in which we can “see without being seen” (Lorenz, 1952), specifically one that conceals us frontally from others, without restricting our own view. It is suggested that these optimal conditions might be translated architecturally as screens, the apertures of which are large enough for a building occupant to see through unobstructed from close by, but small enough to conceal them from the view of someone looking from a distance outside.

Keywords: foreground obstructions, prospection, spatial depth, window views

Procedia PDF Downloads 97
263 Slope Stability and Landslides Hazard Analysis, Limitations of Existing Approaches, and a New Direction

Authors: Alisawi Alaa T., Collins P. E. F.

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The analysis and evaluation of slope stability and landslide hazards are landslide hazards are critically important in civil engineering projects and broader considerations of safety. The level of slope stability risk should be identified due to its significant and direct financial and safety effects. Slope stability hazard analysis is performed considering static and/or dynamic loading circumstances. To reduce and/or prevent the failure hazard caused by landslides, a sophisticated and practical hazard analysis method using advanced constitutive modeling should be developed and linked to an effective solution that corresponds to the specific type of slope stability and landslides failure risk. Previous studies on slope stability analysis methods identify the failure mechanism and its corresponding solution. The commonly used approaches include used approaches include limit equilibrium methods, empirical approaches for rock slopes (e.g., slope mass rating and Q-slope), finite element or finite difference methods, and district element codes. This study presents an overview and evaluation of these analysis techniques. Contemporary source materials are used to examine these various methods on the basis of hypotheses, the factor of safety estimation, soil types, load conditions, and analysis conditions and limitations. Limit equilibrium methods play a key role in assessing the level of slope stability hazard. The slope stability safety level can be defined by identifying the equilibrium of the shear stress and shear strength. The slope is considered stable when the movement resistance forces are greater than those that drive the movement with a factor of safety (ratio of the resistance of the resistance of the driving forces) that is greater than 1.00. However, popular and practical methods, including limit equilibrium approaches, are not effective when the slope experiences complex failure mechanisms, such as progressive failure, liquefaction, internal deformation, or creep. The present study represents the first episode of an ongoing project that involves the identification of the types of landslides hazards, assessment of the level of slope stability hazard, development of a sophisticated and practical hazard analysis method, linkage of the failure type of specific landslides conditions to the appropriate solution and application of an advanced computational method for mapping the slope stability properties in the United Kingdom, and elsewhere through geographical information system (GIS) and inverse distance weighted spatial interpolation(IDW) technique. This study investigates and assesses the different assesses the different analysis and solution techniques to enhance the knowledge on the mechanism of slope stability and landslides hazard analysis and determine the available solutions for each potential landslide failure risk.

Keywords: slope stability, finite element analysis, hazard analysis, landslides hazard

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262 Exogenous Application of Silicon through the Rooting Medium Modulate Growth, Ion Uptake, and Antioxidant Activity of Barley (Hordeum vulgare L.) Under Salt Stress

Authors: Sibgha Noreen, Muhammad Salim Akhter, Seema Mahmood

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Salt stress is an abiotic stress that causes a heavy toll on growth and development and also reduces the productivity of arable and horticultural crops. Globally, a quarter of total arable land has fallen prey to this menace, and more is being encroached because of the usage of brackish water for irrigation purposes. Though barley is categorized as salt-tolerant crop, but cultivars show a wide genetic variability in response to it. In addressing salt stress, silicon nutrition would be a facile tool for enhancing salt tolerant to sustain crop production. A greenhouse study was conducted to evaluate the response of barley (Hordeum vulgare L.) cultivars to silicon nutrition under salt stress. The treatments included [(a) four barley cultivars (Jou-87, B-14002, B-14011, B-10008); (b) two salt levels (0, 200 mM, NaCl); and (c) two silicon levels (0, 200ppm, K2SiO3. nH2O), arranged in a factorial experiment in a completely randomized design with 16 treatments and repeated 4 times. Plants were harvested at 15 days after exposure to different experimental salinity and silicon foliar conditions. Results revealed that various physiological and biochemical attributes differed significantly (p<0.05) in response to different treatments and their interactive effects. Cultivar “B-10008” excelled in biological yield, chlorophyll constituents, antioxidant enzymes, and grain yield compared to other cultivars. The biological yield of shoot and root organs was reduced by 27.3 and 26.5 percent under salt stress, while it was increased by 14.5 and 18.5 percent by exogenous application of silicon over untreated check, respectively. The imposition of salt stress at 200 mM caused a reduction in total chlorophyll content, chl ‘a’ , ‘b’ and ratio a/b by 10.6,16.8,17.1 and 7.1, while spray of 200 ppm silicon improved the quantum of the constituents by 10.4,12.1,10.2,10.3 over untreated check, respectively. The quantum of free amino acids and protein content was enhanced in response to salt stress and the spray of silicon nutrients. The amounts of superoxide dismutase, catalases, peroxidases, hydrogen peroxide, and malondialdehyde contents rose to 18.1, 25.7, 28.1, 29.5, and 17.6 percent over non-saline conditions under salt stress. However, the values of these antioxidants were reduced in proportion to salt stress by 200 ppm silicon applied as rooting medium on barley crops. The salt stress caused a reduction in the number of tillers, number of grains per spike, and 100-grain weight to the amount of 29.4, 8.6, and 15.8 percent; however, these parameters were improved by 7.1, 10.3, and 9.6 percent by foliar spray of silicon over untreated crop, respectively. It is concluded that the barley cultivar “B-10008” showed greater tolerance and adaptability to saline conditions. The yield of barley crops could be potentiated by a foliar spray of 200 ppm silicon at the vegetative growth stage under salt stress.

Keywords: salt stress, silicon nutrition, chlorophyll constituents, antioxidant enzymes, barley crop

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261 An Action Research Study of Developing Foreign Language Teachers’ Intercultural Competence

Authors: Wei Hing Rosenkvist

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In the past few decades, concerns and demands of promoting student intercultural communicative competence in foreign language education have been increasing along with the rapid growth of information technologies and globalization in the 21st century. In Sweden, related concepts such as internationalization, global citizenship, multiculturalism, and intercultural communication etc., are also keywords that would be found in the written learning objectives of the foreign language education in all levels. Being one of the leading higher institutes in distance education in Europe, Dalarna University clearly states that after completion of the teacher education program, students shall understand the needs for integrating internationalization, intercultural and global perspective in teaching and learning in Swedish schools and implement their own studies to promote education in an international and global context. Despite the fact that many teachers and educators agree with the institutes’ mission and vision about the importance of internationalization and the need of increasing student understanding of intercultural and global perspective, they might find this objective unattainable and restricted due to the nature of the subject and their personal knowledge of intercultural competence. When conducting a comprehensive Chinese language course for the students who are going to become Chinese foreign language teachers, the researcher found that all the learning objectives are linguistic oriented while grammatical components dominate the entire course. Apparently, there is a gap between the learning objectives of the course and the DU’s mission of fostering an international learner with intercultural and globalized perspectives. How to include this macro-learning objective in a foreign language course is a great challenge to the educator. Although scholars from different academic domains have provided different theoretical frameworks and approaches for developing student intercultural competence, research that focuses on the didactic perspectives of developing student intercultural competence in teaching Chinese as a foreign language education (CFL) is limited and practical examples are rare. This has motivated the researcher to conduct an action research study that aims at integrating DU’s macro-learning objective in a current CFL course through different didactic practices with a purpose of developing the teacher student intercultural competence. This research study aims to, firstly, illustrate the cross-cultural knowledge integrated into the present Chinese language course for developing intercultural competence. Secondly, it investigates different didactic means that can be utilized to deliver cross-cultural knowledge to student teachers in the present course without generating dramatic disturbance of the syllabus. Thirdly, it examines the effectiveness of these didactic means in enhancing teacher student intercultural competence regarding the need for integrating and implementing internationalization, intercultural and global perspectives in teaching and learning in Swedish schools. Last but not least, it intends to serve as a practical example for developing the student teachers’ intercultural competence in foreign language education in DU and fill in the research gap of this academic domain worldwide.

Keywords: intercultural competence, foreign language education, action research, teacher education

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260 Electrophoretic Light Scattering Based on Total Internal Reflection as a Promising Diagnostic Method

Authors: Ekaterina A. Savchenko, Elena N. Velichko, Evgenii T. Aksenov

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The development of pathological processes, such as cardiovascular and oncological diseases, are accompanied by changes in molecular parameters in cells, tissues, and serum. The study of the behavior of protein molecules in solutions is of primarily importance for diagnosis of such diseases. Various physical and chemical methods are used to study molecular systems. With the advent of the laser and advances in electronics, optical methods, such as scanning electron microscopy, sedimentation analysis, nephelometry, static and dynamic light scattering, have become the most universal, informative and accurate tools for estimating the parameters of nanoscale objects. The electrophoretic light scattering is the most effective technique. It has a high potential in the study of biological solutions and their properties. This technique allows one to investigate the processes of aggregation and dissociation of different macromolecules and obtain information on their shapes, sizes and molecular weights. Electrophoretic light scattering is an analytical method for registration of the motion of microscopic particles under the influence of an electric field by means of quasi-elastic light scattering in a homogeneous solution with a subsequent registration of the spectral or correlation characteristics of the light scattered from a moving object. We modified the technique by using the regime of total internal reflection with the aim of increasing its sensitivity and reducing the volume of the sample to be investigated, which opens the prospects of automating simultaneous multiparameter measurements. In addition, the method of total internal reflection allows one to study biological fluids on the level of single molecules, which also makes it possible to increase the sensitivity and the informativeness of the results because the data obtained from an individual molecule is not averaged over an ensemble, which is important in the study of bimolecular fluids. To our best knowledge the study of electrophoretic light scattering in the regime of total internal reflection is proposed for the first time, latex microspheres 1 μm in size were used as test objects. In this study, the total internal reflection regime was realized on a quartz prism where the free electrophoresis regime was set. A semiconductor laser with a wavelength of 655 nm was used as a radiation source, and the light scattering signal was registered by a pin-diode. Then the signal from a photodetector was transmitted to a digital oscilloscope and to a computer. The autocorrelation functions and the fast Fourier transform in the regime of Brownian motion and under the action of the field were calculated to obtain the parameters of the object investigated. The main result of the study was the dependence of the autocorrelation function on the concentration of microspheres and the applied field magnitude. The effect of heating became more pronounced with increasing sample concentrations and electric field. The results obtained in our study demonstrated the applicability of the method for the examination of liquid solutions, including biological fluids.

Keywords: light scattering, electrophoretic light scattering, electrophoresis, total internal reflection

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259 Numerical Investigation of Flow Boiling within Micro-Channels in the Slug-Plug Flow Regime

Authors: Anastasios Georgoulas, Manolia Andredaki, Marco Marengo

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The present paper investigates the hydrodynamics and heat transfer characteristics of slug-plug flows under saturated flow boiling conditions within circular micro-channels. Numerical simulations are carried out, using an enhanced version of the open-source CFD-based solver ‘interFoam’ of OpenFOAM CFD Toolbox. The proposed user-defined solver is based in the Volume Of Fluid (VOF) method for interface advection, and the mentioned enhancements include the implementation of a smoothing process for spurious current reduction, the coupling with heat transfer and phase change as well as the incorporation of conjugate heat transfer to account for transient solid conduction. In all of the considered cases in the present paper, a single phase simulation is initially conducted until a quasi-steady state is reached with respect to the hydrodynamic and thermal boundary layer development. Then, a predefined and constant frequency of successive vapour bubbles is patched upstream at a certain distance from the channel inlet. The proposed numerical simulation set-up can capture the main hydrodynamic and heat transfer characteristics of slug-plug flow regimes within circular micro-channels. In more detail, the present investigation is focused on exploring the interaction between subsequent vapour slugs with respect to their generation frequency, the hydrodynamic characteristics of the liquid film between the generated vapour slugs and the channel wall as well as of the liquid plug between two subsequent vapour slugs. The proposed investigation is carried out for the 3 different working fluids and three different values of applied heat flux in the heated part of the considered microchannel. The post-processing and analysis of the results indicate that the dynamics of the evolving bubbles in each case are influenced by both the upstream and downstream bubbles in the generated sequence. In each case a slip velocity between the vapour bubbles and the liquid slugs is evident. In most cases interfacial waves appear close to the bubble tail that significantly reduce the liquid film thickness. Finally, in accordance with previous investigations vortices that are identified in the liquid slugs between two subsequent vapour bubbles can significantly enhance the convection heat transfer between the liquid regions and the heated channel walls. The overall results of the present investigation can be used to enhance the present understanding by providing better insight of the complex, underpinned heat transfer mechanisms in saturated boiling within micro-channels in the slug-plug flow regime.

Keywords: slug-plug flow regime, micro-channels, VOF method, OpenFOAM

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258 Phylogenetic Analysis of Georgian Populations of Potato Cyst Nematodes Globodera Rostochiensis

Authors: Dali Gaganidze, Ekaterine Abashidze

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Potato is one of the main agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. In traditional potato growing regions (Svaneti, Samckhet javaheti and Tsalka), the yield is higher than 30-35 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (PCN) are harmful around the world. Yield losses caused by PCN are estimated up to 30%. Rout surveys conducted in two geographically distinct regions of Georgia producing potatoes - Samtskhe - Javakheti and Svaneti revealed potato cyst nematode Globodera rostochiensi. The aim of the study was the Phylogenetic analyses of Globodera rostochiensi revealed in Georgia by the amplification and sequencing of 28S gen in the D3 region and intergenic ITS1-15.8S-ITS2 region. Identification of all the samples from the two Globodera populations (Samtskhe - Javakheti and Svaneti), i.e., G. rostochiensis (20 isolates) were confirmed by conventional multiplex PCR with ITS 5 universal and PITSp4, PITSr3 specific primers of the cyst nematodes’ (G. pallida, G. rostochiensis). The size of PCR fragment 434 bp confirms that PCN samples from two populations, Samtskhe- Javakheti and Svaneti, belong to G. rostochiensi . The ITS1–5.8S-ITS2 regions were amplified using prime pairs: rDNA1 ( 5’ -TTGATTACGTCCCTGCCCTTT-3’ and rDNA2( 5’ TTTCACTCGCCGTTACTAAGG-3’), D3 expansion regions were amplified using primer pairs: D3A (5’ GACCCCTCTTGAAACACGGA-3’) and D3B (5’-TCGGAAGGAACCAGCTACTA-3’. PCR products of each region were cleaned up and sequenced using an ABI 3500xL Genetic Analyzer. Obtained sequencing results were analyzed by computer program BLASTN (https://blast.ncbi.nlm.nih.gov/Blast.cg). Phylogenetic analyses to resolve the relationships between the isolates were conducted in MEGA7 using both distance- and character-based methods. Based on analysis of G.rostochiensis isolate`s D3 expansion regions are grouped in three major clades (A, B and C) on the phylogenetic tree. Clade A is divided into three subclades; clade C is divided into two subclades. Isolates from the Samtckhet-javakheti population are in subclade 1 of clade A and isolates in subclade 1 of clade C. Isolates) from Svaneti populations are in subclade 2 of clade A and in clad B. In Clade C, subclade two is presented by three isolates from Svaneti and by one isolate (GL17) from Samckhet-Javakheti. . Based on analysis of G.rostochiensis isolate`s ITS1–5.8S-ITS2 regions are grouped in two main clades, the first contained 20 Georgian isolates of Globodera rostochiensis from Svaneti . The second clade contained 15 isolates of Globodera rostochiensis from Samckhet javakheti. Our investigation showed of high genetic variation of D3 and ITS1–5.8S-ITS2 region of rDNA of the isolates of G. rostochiensis from different geographic origins (Svameti, Samckhet-Javakheti) of Georgia. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia : Project # FR17_235

Keywords: globodera rostochiensi, PCR, phylogenetic tree, sequencing

Procedia PDF Downloads 175
257 An Investigation of Tetraspanin Proteins’ Role in UPEC Infection

Authors: Fawzyah Albaldi

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Urinary tract infections (UTIs) are the most prevalent of infectious diseases and > 80% are caused by uropathogenic E. coli (UPEC). Infection occurs following adhesion to urothelial plaques on bladder epithelial cells, whose major protein constituent are the uroplakins (UPs). Two of the four uroplakins (UPIa and UPIb) are members of the tetraspanin superfamily. The UPEC adhesin FimH is known to interact directly with UPIa. Tetraspanins are a diverse family of transmembrane proteins that generally act as “molecular organizers” by binding different proteins and lipids to form tetraspanin enriched microdomains (TEMs). Previous work by our group has shown that TEMs are involved in the adhesion of many pathogenic bacteria to human cells. Adhesion can be blocked by tetraspanin-derived synthetic peptides, suggesting that tetraspanins may be valuable drug targets. In this study, we investigate the role of tetraspanins in UPEC adherence to bladder epithelial cells. Human bladder cancer cell lines (T24, 5637, RT4), commonly used as in-vitro models to investigate UPEC infection, along with primary human bladder cells, were used in this project. The aim was to establish a model for UPEC adhesion/infection with the objective of evaluating the impact of tetraspanin-derived reagents on this process. Such reagents could reduce the progression of UTI, particularly in patients with indwelling catheters. Tetraspanin expression on the bladder cells was investigated by q-PCR and flow cytometry, with CD9 and CD81 generally highly expressed. Interestingly, despite these cell lines being used by other groups to investigate FimH antagonists, uroplakin proteins (UPIa, UPIb and UPIII) were poorly expressed at the cell surface, although some were present intracellularly. Attempts were made to differentiate the cell lines, to induce cell surface expression of these UPs, but these were largely unsuccessful. Pre-treatment of bladder epithelial cells with anti-CD9 monoclonal antibody significantly decreased UPEC infection, whilst anti-CD81 had no effects. A short (15aa) synthetic peptide corresponding to the large extracellular region (EC2) of CD9 also significantly reduced UPEC adherence. Furthermore, we demonstrated specific binding of that fluorescently tagged peptide to the cells. CD9 is known to associate with a number of heparan sulphate proteoglycans (HSPGs) that have also been implicated in bacterial adhesion. Here, we demonstrated that unfractionated heparin (UFH)and heparin analogs significantly inhibited UPEC adhesion to RT4 cells, as did pre-treatment of the cells with heparinases. Pre-treatment with chondroitin sulphate (CS) and chondroitinase also significantly decreased UPEC adherence to RT4 cells. This study may shed light on a common pathogenicity mechanism involving the organisation of HSPGs by tetraspanins. In summary, although we determined that the bladder cell lines were not suitable to investigate the role of uroplakins in UPEC adhesion, we demonstrated roles for CD9 and cell surface proteoglycans in this interaction. Agents that target these may be useful in treating/preventing UTIs.

Keywords: UTIs, tspan, uroplakins, CD9

Procedia PDF Downloads 85