Search results for: Molecular modeling of 17-picolyl and 17-picolinylidene androstane derivatives with anticancer activity
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Search results for: Molecular modeling of 17-picolyl and 17-picolinylidene androstane derivatives with anticancer activity

2235 Evaluation of Goji By-Product as a Value-Added Ingredient for the Functional Food Industry

Authors: Sanaa Ragaee, Paragyani Bora, Wee Teng Tan, Xin Hu

Abstract:

Goji berry (Lycium barbarum) is a member of the family Solanaceae which is grown widely in China, Tibet, and other parts of Asia. Its fruits are 1–2 cm-long, bright orange-red ellipsoid berries and it has a long tradition as a food and medicinal plant. Goji berries are believed to boost immune system properties. The berries are considered an excellent source of macronutrients, micronutrients, vitamins, minerals and several bioactive components. Studies have shown effects of goji fruit on aging, neuroprotection, general well-being, fatigue/endurance, metabolism/energy expenditure, glucose control in diabetics and glaucoma, antioxidant properties, immunomodulation and anti-tumor activity. Goji berries are being used to prepare Goji beverage, and the remaining solid material is considered as by-product. The by-product is currently unused and disposed as waste despite its potential as a value-added food ingredient. Therefore, this study is intended to evaluate nutritional properties of Goji by-product and its potential applications in the baking industry. The Goji by-product was freeze dried and ground to pass through 1 mm screen prior to evaluation and food use. The Goji by-product was found to be a rich source of fiber (54%) and free phenolic components (1,307 µg/g), protein (13.6%), ash (3.3%) and fat (10%). Incorporation of the Goji by-product in muffins and cookies at various levels (10-40%) significantly improved the nutritional quality of the baked products. The baked products were generally accepted and highly rated by panelists at 20% replacement level. The results indicate the potential of Goji by-product as a value-added ingredient in particular as a source of dietary fiber and protein.

Keywords: Goji, by-product, phenolics, fibers, baked products

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2234 Purification of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) from Fish Oil Using HPLC Method and Investigation of Their Antibacterial Effects on Some Pathogenic Bacteria

Authors: Yılmaz Uçar, Fatih Ozogul, Mustafa Durmuş, Yesim Ozogul, Ali Rıza Köşker, Esmeray Kuley Boğa, Deniz Ayas

Abstract:

The aim of this study was to purified eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), that are essential oils from trout oil, using high-performance liquid chromatography (HPLC) method, bioconverted EPA and DHA into bioconverted EPA (bEPA), bioconverted DHA (bDHA) extracts by P. aeruginosa PR3. Moreover, in vitro antibacterial activity of bEPA and bDHA was investigated using disc diffusion methods and minimum inhibitory concentration (MIC). EPA and DHA concentration of 11.1% and 15.9% in trout oil increased in 58.64% and 40.33% after HPLC optimisation, respectively. In this study, EPA and DHA enriched products were obtained which are to be used as valuable supplements for food and pharmaceutical purposes. The bioconverted EPA and DHA exhibited antibacterial activities against two Gram-positive bacteria (Listeria monocytogenes ATCC 7677 and Staphylococcus aureus ATCC 29213) and six Gram-negative bacteria (Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC700603, Enterococcus faecalis ATCC 29212, Aeromonas hydrophila NCIMB 1135, and Salmonella Paratyphi A NCTC 13). Inhibition zones and MIC value of bEPA and bDHA against bacterial strains ranged from 7 to 12 mm and from 350 to 2350 μg/mL, respectively. Our results suggested that the crude extracts of bioconversion of EPA and DHA by P. aeruginosa PR3 can be considered as promising antimicrobials in improving food safety by controlling foodborne pathogens.

Keywords: High-Performance Liquid Chromatography (HPLC), docosahexaenoic acid, DHA, eicosapentaenoic acid, EPA, minimum inhibitory concentration, MIC, Pseudomonas aeruginosa PR3

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2233 Development of Power System Stability by Reactive Power Planning in Wind Power Plant With Doubley Fed Induction Generators Generator

Authors: Mohammad Hossein Mohammadi Sanjani, Ashknaz Oraee, Oriol Gomis Bellmunt, Vinicius Albernaz Lacerda Freitas

Abstract:

The use of distributed and renewable sources in power systems has grown significantly, recently. One the most popular sources are wind farms which have grown massively. However, ¬wind farms are connected to the grid, this can cause problems such as reduced voltage stability, frequency fluctuations and reduced dynamic stability. Variable speed generators (asynchronous) are used due to the uncontrollability of wind speed specially Doubley Fed Induction Generators (DFIG). The most important disadvantage of DFIGs is its sensitivity to voltage drop. In the case of faults, a large volume of reactive power is induced therefore, use of FACTS devices such as SVC and STATCOM are suitable for improving system output performance. They increase the capacity of lines and also passes network fault conditions. In this paper, in addition to modeling the reactive power control system in a DFIG with converter, FACTS devices have been used in a DFIG wind turbine to improve the stability of the power system containing two synchronous sources. In the following paper, recent optimal control systems have been designed to minimize fluctuations caused by system disturbances, for FACTS devices employed. For this purpose, a suitable method for the selection of nine parameters for MPSH-phase-post-phase compensators of reactive power compensators is proposed. The design algorithm is formulated ¬¬as an optimization problem searching for optimal parameters in the controller. Simulation results show that the proposed controller Improves the stability of the network and the fluctuations are at desired speed.

Keywords: renewable energy sources, optimization wind power plant, stability, reactive power compensator, double-feed induction generator, optimal control, genetic algorithm

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2232 The Term of Intellectual Property and Artificial Intelligence

Authors: Yusuf Turan

Abstract:

Definition of Intellectual Property Rights according to the World Intellectual Property Organization: " Intellectual property (IP) refers to creations of the mind, such as inventions; literary and artistic works; designs; and symbols, names and images used in commerce." It states as follows. There are 2 important points in the definition; we can say that it is the result of intellectual activities that occur by one or more than one PERSON and as INNOVATION. When the history and development of the relevant definitions are briefly examined, it is realized that these two points have remained constant and Intellectual Property law and rights have been shaped around these two points. With the expansion of the scope of the term Intellectual Property as a result of the development of technology, especially in the field of artificial intelligence, questions such as "Can "Artificial Intelligence" be an inventor?" need to be resolved within the expanding scope. In the past years, it was ruled that the artificial intelligence named DABUS seen in the USA did not meet the definition of "individual" and therefore would be an inventor/inventor. With the developing technology, it is obvious that we will encounter such situations much more frequently in the field of intellectual property. While expanding the scope, we should definitely determine the boundaries of how we should decide who performs the mental activity or creativity that we call indispensable on the inventor/inventor according to these problems. As a result of all these problems and innovative situations, it is clearly realized that not only Intellectual Property Law and Rights but also their definitions need to be updated and improved. Ignoring the situations that are outside the scope of the current Intellectual Property Term is not enough to solve the problem and brings uncertainty. The fact that laws and definitions that have been operating on the same theories for years exclude today's innovative technologies from the scope contradicts intellectual property, which is expressed as a new and innovative field. Today, as a result of the innovative creation of poetry, painting, animation, music and even theater works with artificial intelligence, it must be recognized that the definition of Intellectual Property must be revised.

Keywords: artificial intelligence, innovation, the term of intellectual property, right

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2231 In vivo Antidiarrheal and ex-vivo Spasmolytic Activities of the Aqueous Extract of the Roots of Echinops kebericho Mesfin in Rodents and Isolated Guinea-Pig Ileum

Authors: Fisseha Shiferie (Bpharm, Mpharm)

Abstract:

Diarrhea is a common gastrointestinal disorder characterized by an increase in stool frequency and a change in stool consistency. Inspite of the availability of many drugs as antidiarrheal agents, the search for a drug with affordable cost and better efficacy is essential to overcome diarrheal problems. The root extract of Echinops kebericho, is used by traditional practitioners for the treatment of diarrhea. However, the scientific basis for this usage has not been yet established. The purpose of the present study was to evaluate the antidiarrheal and spasmolytic activities of the aqueous extract of the roots of E. kebericho in rodents and isolated guinea-pig ileum preparations. In the castor oil induced intestinal transit test, E. kebericho produced a significant (p < 0.01) dose dependent decrease in propulsion with peristaltic index values of 45.05±3.3, 42.71±2.25 and 33.17±3.3%, respectively at doses of 100, 200 and 400 mg/kg compared with 63.43±7.3% for control. In the castor oil-induced diarrhea test, the mean defecation was reduced from 1.81±0.18 to 0.99 ± 0.21 compared with 2.59 ±0.81 for control. The extract (at doses stated above) significantly decreased the volume of intestinal fluid secretion induced by castor oil (2.31±0.1 to 2.01±0.2) in relation to 3.28±0.3 for control. When tested on a guinea-pig ileum, root extract of Echinops kebericho exhibited a dose dependent spasmolytic effect, 23.07 % being its highest inhibitory effect. The results obtained in this study give some scientific support to the use of Echinops kebericho as an antidiarrheal agent due to its inhibitory effects on the different diarrheal parameters used in this study.

Keywords: antidiarrheal activity, E. kebericho, traditional medicine, diarrhea, enteropooling, and intestinal transit

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2230 Effects of Planned Pre-laboratory Discussion on Physics Students’ Acquisition of Science Process Skills in Kontagora, Niger State

Authors: Akano Benedict Ubawuike

Abstract:

This study investigated the effects of pre-laboratory discussion on physics students’ acquisition of science process skills. The study design was quasi-experimental and purposive sampling technique was applied in selecting two schools in Kontagora Town for the research based on the availability of a good physics laboratory. Intact classes already grouped by the school for the sake of small laboratory space and equipment, comprising Thirty (30) students, 15 for experimental group in School A and 15 for control in school B were the subjects for the research. The instrument used for data collection was the lesson prepared for pre – practical discussion and researcher made Science Process Skill Test (SPST ) and two (2) research questions, and two (2) research hypotheses were developed to guide the study. The data collected were analyzed using means and t-Test statistics at 0.05 level of significance. The study revealed that pre-laboratory discussion was found to be more efficacious in enhancing students’ acquisition of science process skills. It also revealed that gender, had no significant effect on students’ acquisition of science process skills. Based on the findings, it was recommended among others that teachers should encourage students to develop interest in practical activities by engaging them in pre-laboratory discussion and providing instructional materials that will challenge them to be actively involved during practical lessons. It is also recommended that Ministries of Education and professional organizations like Science Teachers' Association of Nigeria (STAN) should organize workshops, seminars and conferences for physics teachers and Physics concepts should be taught with practical activity so that the students will do science instead of learning about science.

Keywords: physics, laboratory, discussion, students, acquisition, science process skills

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2229 Rebamipide Retards CCL4 Induced Hepatic Fibrosis: A Role of PGE2

Authors: Alaa E. El-sisi, Sherin Zakaria

Abstract:

Rebamipide is an antiulcer drug with unique properties such as anti-inflammatory action. It induces endogenous prostaglandin e2 (PGE2). PGE2 is considered as a potent physiological suppressor of liver fibrosis. Aim of study: This study investigated the effect of rebamipide on hepatic fibrosis. Material and Method: Hepatic fibrosis was induced by intraperitoneal injections (IP) injection of CCl4 (0.45 mL/kg) in corn oil 1:5 twice a week for 4 weeks. Rats were divided into four groups as follow: Group 1 treated with CCL4 only, group 2 and 3 treated with CCL4 and rebamipide 60 mg/kg/day (group2) or 100 mg/kg/day (group3), and the fourth group was considered as control group and treated with vehicles. ALT, AST, and Bilirubin were assayed in serum. Antioxidant markers such as malondialdhyde (MDA) and superoxide dismutase (SOD) and fibrotic markers such as hyaluronic acid (HA) and procollagen-III (procol-III) were evaluated in liver tissues. IL-10 as well as PGE2 were also assayed in liver tissues. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen precipitation in liver tissues was visualized using masson trichrom stain. Results: Rebamipide inhibit CCL4 induced increase in ALT and AST significantly (p < 0.05). Rebamipide exerted an antioxidant effect as it inhibits CCL4 induced increased MDA level and decreased SOD activity. Fibrotic markers assay revealed that repamipide (60 or 100 mg/kg/day) decreased the level of procol-III and HA compared to CCl4 (p < 0.05). Oral administration of Rebamipide was associated with a significant increase (p < 0.05) of PGE2 and IL-10. Rebamipide especially at the dose of (100 mg/kg/day) restores liver histology structure and abolish collagen precipitation in liver tissues. Conclusion: Rebamipide retards hepatic fibrosis induced by CCL4 may be through the induction of PGE2 level.

Keywords: fibrotic markers, hepatic fibrosis, PGE2, rebamipide

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2228 Comparison of Inexpensive Cell Disruption Techniques for an Oleaginous Yeast

Authors: Scott Nielsen, Luca Longanesi, Chris Chuck

Abstract:

Palm oil is obtained from the flesh and kernel of the fruit of oil palms and is the most productive and inexpensive oil crop. The global demand for palm oil is approximately 75 million metric tonnes, a 29% increase in global production of palm oil since 2016. This expansion of oil palm cultivation has resulted in mass deforestation, vast biodiversity destruction and increasing net greenhouse gas emissions. One possible alternative is to produce a saturated oil, similar to palm, from microbes such as oleaginous yeast. The yeasts can be cultured on sugars derived from second-generation sources and do not compete with tropical forests for land. One highly promising oleaginous yeast for this application is Metschnikowia pulcherrima. However, recent techno-economic modeling has shown that cell lysis and standard lipid extraction are major contributors to the cost of the oil. Typical cell disruption techniques to extract either single cell oils or proteins have been based around bead-beating, homogenization and acid lysis. However, these can have a detrimental effect on lipid quality and are energy-intensive. In this study, a vortex separator, which produces high sheer with minimal energy input, was investigated as a potential low energy method of lysing cells. This was compared to four more traditional methods (thermal lysis, acid lysis, alkaline lysis, and osmotic lysis). For each method, the yeast loading was also examined at 1 g/L, 10 g/L and 100 g/L. The quality of the cell disruption was measured by optical cell density, cell counting and the particle size distribution profile comparison over a 2-hour period. This study demonstrates that the vortex separator is highly effective at lysing the cells and could potentially be used as a simple apparatus for lipid recovery in an oleaginous yeast process. The further development of this technology could potentially reduce the overall cost of microbial lipids in the future.

Keywords: palm oil substitute, metschnikowia pulcherrima, cell disruption, cell lysis

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2227 A Methodology for the Identification of Technological Gaps and the Measurement of the Level of Technological Acceptance in the Rural Sector in Colombia

Authors: Anyi Katherine Garzon Robles, Luis Carlos Gomez Florez

Abstract:

Since the advent of the Internet, the use of Information Technologies (IT) has increased exponentially. The field of informatics and telecommunications has put on the table countless possibilities for the development of different socio-economic activities, promoting a change of social paradigm and the emergence of the so-called information and knowledge society. For more than a decade, the Colombian government has been working on the incorporation of IT into the public sector through an e-government strategy. However, to date, many technological gaps has not yet been identified in the country to our knowledge, especially in rural areas and far from large cities, where factors such as low investment and the expansion of the armed conflict have led to economic and technological stagnation. This paper presents the research results obtained from the execution of a research project, which was approach from a qualitative approach and a methodological design of a participatory action research type. This design consists of nine fundamental stages divided into four work cycles. For which different strategies for data collection and analysis were established. From which, a methodology was obtained for the identification of technological gaps and the measurement of the level of technological acceptance in the rural sector, based on the TAM (Technology Acceptance Model) model, as a previous activity to the development of IT solutions framed in the e-government strategy in Colombia. The result of this research work represents a contribution from academia for the improvement of the country's technological development and a guide for the proper planning of IT solutions aimed at promoting a close relationship between government and citizens.

Keywords: E-government, knowledge society, level of technological acceptance, technological gaps, technology acceptance model

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2226 Monitoring Prospective Sites for Water Harvesting Structures Using Remote Sensing and Geographic Information Systems-Based Modeling in Egypt

Authors: Shereif. H. Mahmoud

Abstract:

Egypt has limited water resources, and it will be under water stress by the year 2030. Therefore, Egypt should consider natural and non-conventional water resources to overcome such a problem. Rain harvesting is one solution. This Paper presents a geographic information system (GIS) methodology - based on decision support system (DSS) that uses remote sensing data, filed survey, and GIS to identify potential RWH areas. The input into the DSS includes a map of rainfall surplus, slope, potential runoff coefficient (PRC), land cover/use, soil texture. In addition, the outputs are map showing potential sites for RWH. Identifying suitable RWH sites implemented in the ArcGIS model environment using the model builder of ArcGIS 10.1. Based on Analytical hierarchy process (AHP) analysis taking into account five layers, the spatial extents of RWH suitability areas identified using Multi-Criteria Evaluation (MCE). The suitability model generated a suitability map for RWH with four suitability classes, i.e. Excellent, Moderate, Poor, and unsuitable. The spatial distribution of the suitability map showed that the excellent suitable areas for RWH concentrated in the northern part of Egypt. According to their averages, 3.24% of the total area have excellent and good suitability for RWH, while 45.04 % and 51.48 % of the total area are moderate and unsuitable suitability, respectively. The majority of the areas with excellent suitability have slopes between 2 and 8% and with an intensively cultivated area. The major soil type in the excellent suitable area is loam and the rainfall range from 100 up to 200 mm. Validation of the used technique depends on comparing existing RWH structures locations with the generated suitability map using proximity analysis tool of ArcGIS 10.1. The result shows that most of exiting RWH structures categorized as successful.

Keywords: rainwater harvesting (RWH), geographic information system (GIS), analytical hierarchy process (AHP), multi-criteria evaluation (MCE), decision support system (DSS)

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2225 Willingness of Spanish Wineries to Implement Renewable Energies in Their Vineyards and Wineries, as Well as the Limitations They Perceive for Their Implementation

Authors: Javier Carroquino, Nieves García-Casarejos, Pilar Gargallo

Abstract:

Climate change, depletion of non-renewable resources in the current energies, pollution from them, the greater ecological awareness of the population, are factors that suggest the change of energy sources in business. The agri-food industry is a growth sector, concerned about product innovation, process and with a clear awareness of what climate change may mean for it. This sector is supposed to have a high receptivity to the implementation of clean energy, as this favors not only the environment but also the essence of its business. This work, through surveys, aims to know the willingness of Spanish wineries to implement renewable energies in their vineyards, as well as the limitations they perceive for their implementation. This questionnaire allows the characterization of the sector in terms of its geographical typologies, their activity levels, their perception of environmental issues, the degree of implementation of measures to mitigate climate change and improve energy efficiency, and its uses and energy consumption. The analysis of data proves that the penetration of renewable energies is still at low levels, being the most used energies, solar thermal, photovoltaic and biomass. The initial investment seems to be at the origin of the lack of implantation of this type of energy in the wineries, and not so much the costs of operations and maintenance. The environmental management of the wineries is still at an embryonic stage within the company's organization chart, because these services are either outsourced or, if technicians are available, they are not exclusively dedicated to these tasks. However, there is a strong environmental awareness, as evidenced by the number of climate change mitigation and energy efficiency measures already adopted. The gap between high awareness and low achievement is probably due to the lack of knowledge about how to do it or the perception of a high cost.

Keywords: survey, renewable energy, winery, Spanish case

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2224 Dynamics of Parent to Adolescent Communication on Sexual and Reproductive Health in Sub-Saharan Africa: A Focus on Barriers and Policy Implications

Authors: Douglas Nyathi, Mxolisi Sibanda, Joram Ndlovuu, Thulani Dube, Innocent T. Mahiya

Abstract:

Communication of sexual matters between the parents and adolescents has been seen as one of the strategies that could play a cardinal role in encouraging adolescents to be responsible and delay sexual debut or avoid unprotected sexual intercourse. The increasing rate of teenage pregnancies and new HIV/AIDS infections among adolescents in Sub-Saharan Africa makes the phenomenon worth analysis. The purpose of this paper is to interrogate the dynamics of parent-adolescent communication on sexual and reproductive health in Sub-Sahara. Specifically the paper focuses on barriers to communication between parents and adolescents on sexual and reproductive health and its policy implications. It emanates from the paper that communication on sexual and reproductive health at household level is triggered by death of a relative from a sexual related illness, suspicion on sexual activity, radio programmes and in some instances fliers. Literature engagement reveals that communication between parents and adolescents on sexual and reproductive health is made difficult by economic factors (poverty, lack of privacy and low self-esteem), household demographics (age, sex, class, death), socio-cultural factors (beliefs and religious values) as well as social media. We argue that there is need to use broadcast mediato come up with radio and television programmes that create family environments in which sexual and reproductive health issues are discussed. We also recommend that government departments and Non-Governmental Organisations concerned with sexuality issues need to undertake studies that can help dismantle taboos, prejudices and stereotypes that impede sexual and reproductive health communication between parents and adolescents.

Keywords: parent, adolecsent, communication, sexual and reproductive health

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2223 Drawings Reveal Beliefs of Japanese University Students

Authors: Sakae Suzuki

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Although Japanese students study English for six years in secondary schools, they demonstrate little success with it when they enter higher education. Learners’ beliefs can predict the future behavior of students, so it may be effective to investigate how learners’ beliefs limit their success and how beliefs might be nudged in a positive direction. While many researchers still depend on a questionnaire called BALLI to reveal explicit beliefs, alternative approaches, especially those designed to reveal implicit beliefs, might be helpful for promoting learning. The present study seeks to identify beliefs with a discursive approach using visual metaphors and narratives. Employing a sociocultural framework, this study investigates how students’ beliefs are revealed by drawings of themselves and their surrounding environments and artifacts while they are engaged in language learning. Research questions are: (1) Can we identify beliefs through an analysis of students’ visual narratives? (2) What environments and artifacts can be found in students’ drawings, and what do they mean? (3) To what extent do students see language learning as a solitary, rather than a social, activity? Participants are university students majoring in science and technology in Japan. The questionnaire was administered to 70 entering students in April, 2014. Data included students drawings of themselves as learners of English as well as written descriptions of students’ backgrounds, English-learning experiences, and analogies and metaphors that they used in written descriptions of themselves as learners. Data will be analyzed qualitatively and quantitatively. Anticipated results include students’ perceptions of themselves as language learners, including their sense of agency, awareness of artifacts, and social contexts of language learning. Comments will be made on implications for teaching, as well as the use of visual narratives as research tools, and recommended further research.

Keywords: drawings, learners' beliefs, metaphors, BALLI

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2222 Pavement Management for a Metropolitan Area: A Case Study of Montreal

Authors: Luis Amador Jimenez, Md. Shohel Amin

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Pavement performance models are based on projections of observed traffic loads, which makes uncertain to study funding strategies in the long run if history does not repeat. Neural networks can be used to estimate deterioration rates but the learning rate and momentum have not been properly investigated, in addition, economic evolvement could change traffic flows. This study addresses both issues through a case study for roads of Montreal that simulates traffic for a period of 50 years and deals with the measurement error of the pavement deterioration model. Travel demand models are applied to simulate annual average daily traffic (AADT) every 5 years. Accumulated equivalent single axle loads (ESALs) are calculated from the predicted AADT and locally observed truck distributions combined with truck factors. A back propagation Neural Network (BPN) method with a Generalized Delta Rule (GDR) learning algorithm is applied to estimate pavement deterioration models capable of overcoming measurement errors. Linear programming of lifecycle optimization is applied to identify M&R strategies that ensure good pavement condition while minimizing the budget. It was found that CAD 150 million is the minimum annual budget to good condition for arterial and local roads in Montreal. Montreal drivers prefer the use of public transportation for work and education purposes. Vehicle traffic is expected to double within 50 years, ESALS are expected to double the number of ESALs every 15 years. Roads in the island of Montreal need to undergo a stabilization period for about 25 years, a steady state seems to be reached after.

Keywords: pavement management system, traffic simulation, backpropagation neural network, performance modeling, measurement errors, linear programming, lifecycle optimization

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2221 The Localization and Function of p38α Mitogen-Activated Protein Kinase (MAPK) in Rat Oocytes

Authors: Shifu Hu, Qiong Yu, Wei Xia, Changhong Zhu

Abstract:

Background: P38α MAPK, which is a member of the canonical MAPK family, is activated in response to various extracellular stresses and plays a role in multiple cellular processes. It is well known that p38α MAPK play vital roles in oocyte maturation, but the localization and functional roles of p38α MAPK during the meiotic maturation of rat oocytes remain unknown. Study Design: In this study, western-blot and immunofluorescent staining were used to investigate the expression and subcellular localization of p38α MAPK during the meiotic maturation of rat oocytes. SB203580, a specific inhibitor of p38α MAPK, was used to study the roles of p38α MAPK in the meiotic cell cycle of rat oocytes. Results: The results found that p38α MAPK phosphorylation (p-p38α MAPK, indicative of p38α MAPK activation) was low at the germinal vesicle (GV) stage, increased 3 h after germinal vesicle breakdown (GVBD), and maintained its maximum at MI (metaphase I) or M II (metaphase II). The p-p38α MAPK mainly accumulated in the germinal vesicle and had no obvious expression in the nucleus. From GVBD to M II, p-p38α MAPK was distributed in the cytoplasm around either the chromosomes or the spindle. We used SB203580, an inhibitor of p38α MAPK, to investigate the possible functional role of p38α MAPK during rat oocyte meiotic maturation. Treatment of GV stage oocytes with 20 μM SB203580 blocked p-p38α MAPK activity, and the spindles appeared abnormal. Additionally, the rate of GVBD after 3h of culture with 20 μM SB203580 (58.8%) was significantly inhibited compared with the control (82.5%, p < 0.05), and the polar body extrusion rate after 12 h of culture with SB203580 was also significantly decreased compared with the control (40.1 vs. 73.3%, p < 0.05). Conclusions: These data indicate that p38α MAPK may play a vital role in rat oocyte meiotic maturation.

Keywords: meiotic maturation, oocyte, p38α MAPK, spindle

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2220 Biocontrol of Fusarium Crown and Root Rot and Enhancement of Tomato Solanum lycopersicum L. Growth Using Solanum linnaeanum L. Extracts

Authors: Ahlem Nefzi, Rania Aydi Ben Abdallah, Hayfa Jabnoun-Khiareddine, Nawaim Ammar, Sined Medimagh-Saidana, Mejda Daami-Remadi

Abstract:

In the present study, leaf, stem, and fruit aqueous extracts of native wild Solanum linnaeanum L. were screened for their ability to suppress Fusarium Crown and Root Rot disease and to enhance tomato (Solanum lycopersicum L.) growth under greenhouse conditions. Leaf extract used at 30% w/v was the most effective in reducing leaf and root damage index by 92.3% and the extent of vascular discoloration by 97.56% compared to Fusarium oxyxporum f. sp radicis lycopersici -inoculated and untreated control. A significant promotion of growth parameters (root length, shoot height, root and shoot biomass and stem diameter) was recorded on tomato cv. Rio Grande seedlings by 40.3-94.1% as compared to FORL inoculated control and by 9.6-88.8% over pathogen-free control. All S. linnaeanum aqueous extracts tested significantly stimulated the germination by 10.2 to 80.1% relative to the untreated control. FORL mycelial growth, assessed using the poisoned food technique, varied depending on plant organs, extracts, and concentrations used. Butanolic extracts were the most active, leading to 60.81% decrease in FORL mycelial growth. HPLC analysis of butanolic extract revealed the presence of thirteen phenolic compounds. Thus, S. linnaeanum can be explored as a potential natural source of antifungal and biofertilizing compounds.

Keywords: antifungal activity, HPLC-MS analysis, Fusarium oxysporum f. sp. radicis-lycopersici, tomato growth

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2219 The Moving and Special Ability Influence Player Preference in the Dual Protagonist Game

Authors: Shih-Chieh Liao, Jen-Ying Ma

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Dual protagonists game always bring a unique experience compared to the other games. This research wants to discuss whether the dual protagonists have the moving ability and special ability or not; it will affect the preference of the players. This research will focus on the single-player dual protagonists game. After the observation, we found that when players control the dual protagonists, the moving ability and special ability are a great point defining the preference of players. When players control the character, which is lack of moving ability, they often feel impatient with the inconvenient mechanism and then reduce the will to play with the character or even the game. Furthermore, the special ability is also important in the situation that there is another character to compare with. When the character is too powerful, players tend not to use the weaker one. In addition, gender is a big deal in the games. It surprisingly controls the will of play occasionally. In view of these, this research makes a single-player dual protagonists game and the dual protagonists are limited to male and female. The experiment content detected with Electrodermal Activity (EDA) includes seven different situations. (1) male and females both have the moving ability and special ability. (2) male and female both have a special ability, but female does not have the moving ability. (3) male and females both have a special ability, but the male does not have the moving ability. (4) male and female both have the moving ability, but the male does not have special ability (5) male and female both have the moving ability, but female does not have a special ability (6) male-only has the moving ability and female-only has a special ability (7) male-only has a special ability and female only has the moving ability. The experiment will evaluate the emotional changes of the subjects in those situations. The result sorted by the significance of player preference is (6)>(4)>(1)>(2)>(5)>(3)>(7). The result demonstrates that players prefer females with special abilities or males with moving abilities. The game developer could design the ability of dual protagonists based on this research. Therefore, players may have a better experience.

Keywords: biofeedback, dual protagonists, emotional responses, psychology, user experience

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2218 The Effect of Oil Pollution on Marine Microbial Populations in Israeli Coastal Waters

Authors: Yael Shai, Dror L. Angel, Dror Zurel, Peleg Astrahan, Maxim Rubin-Blum, Eyal Rahav

Abstract:

The high demand for oil and its by-products is symptomatic of the 21st century and occasionally leads to oil spills and pollution of coastal waters. Marine oil pollution may originate from a variety of sources -urban runoff, tanker cleaning, drilling activities, and oil spills. These events may release large amounts of highly toxic polycyclic aromatic hydrocarbons (PAHs) and other pollutants to coastal water, thereby threatening local marine life. Here, we investigated the effects of crude oil on the temporal dynamics of phytoplankton and heterotrophic bacteria in Israeli coastal waters. To this end, we added crude oil (500 µm thick layer, with and without additional nutrients; NO₃ and PO₄) to mesocosms (1m³ bags) containing oligotrophic surface coastal water collected near Haifa during summer and winter. Changes in phytoplankton biomass, activity and diversity were monitored daily for 5-6 days. Our results demonstrate that crude oil addition resulted in a pronounced decrease in phytoplankton biomass and production rates, while heterotrophic bacterial production increased significantly. Importantly, a few days post addition we found that the oil-degrading bacteria, Oleibacter sp. and Oleispira sp. appeared in the mesocosms and that the addition of nutrients (along with the crude oil) further increased this trend. This suggests that oil-degrading bacteria may be NO₃ and PO₄ limited in Israeli coastal waters. The results of this study should enable us to establish improved science-based environmental policy with respect to handling crude oil pollution in this region.

Keywords: heterotrophic bacteria, nutrients, mesocosm, oil pollution, oligotrophic, phytoplankton

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2217 House Facades and Emotions: Exploring the Psychological Impact of Architectural Features

Authors: Nour Tawil, Sandra Weber, Kirsten K. Roessler, Martin Mau, Simone Kuhn

Abstract:

The link between “quality” residential environments and human health and well-being has long been proposed. While the physical properties of a sound environment have been fairly defined, little focus has been given to the psychological impact of architectural elements. Recently, studies have investigated the response to architectural parameters, using measures of physiology, brain activity, and emotion. Results showed different aspects of interest: detailed and open versus blank and closed facades, patterns in perceiving different elements, and a visual bias for capturing faces in buildings. However, in the absence of a consensus on methodologies, the available studies remain unsystematic and face many limitations regarding the underpinning psychological mechanisms. To bridge some of these gaps, an online study was launched to investigate design features that influence the aesthetic judgement and emotional evaluation of house facades, using a well-controlled stimulus set of Canadian houses. A methodical modelling of design features will be performed to extract both high and low level image properties, in addition to segmentation of layout-related features. 300 participants from Canada, Denmark, and Germany will rate the images on twelve psychological dimensions representing appealing aspects of a house. Subjective ratings are expected to correlate with specific architectural elements while controlling for typicality and familiarity, and other individual differences. With the lack of relevant studies, this research aims to identify architectural elements of beneficial qualities that can inform design strategies for optimized residential spaces.

Keywords: architectural elements, emotions, psychological response, residential facades.

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2216 [Keynote Talk]: Mathematical and Numerical Modelling of the Cardiovascular System: Macroscale, Mesoscale and Microscale Applications

Authors: Aymen Laadhari

Abstract:

The cardiovascular system is centered on the heart and is characterized by a very complex structure with different physical scales in space (e.g. micrometers for erythrocytes and centimeters for organs) and time (e.g. milliseconds for human brain activity and several years for development of some pathologies). The development and numerical implementation of mathematical models of the cardiovascular system is a tremendously challenging topic at the theoretical and computational levels, inducing consequently a growing interest over the past decade. The accurate computational investigations in both healthy and pathological cases of processes related to the functioning of the human cardiovascular system can be of great potential in tackling several problems of clinical relevance and in improving the diagnosis of specific diseases. In this talk, we focus on the specific task of simulating three particular phenomena related to the cardiovascular system on the macroscopic, mesoscopic and microscopic scales, respectively. Namely, we develop numerical methodologies tailored for the simulation of (i) the haemodynamics (i.e., fluid mechanics of blood) in the aorta and sinus of Valsalva interacting with highly deformable thin leaflets, (ii) the hyperelastic anisotropic behaviour of cardiomyocytes and the influence of calcium concentrations on the contraction of single cells, and (iii) the dynamics of red blood cells in microvasculature. For each problem, we present an appropriate fully Eulerian finite element methodology. We report several numerical examples to address in detail the relevance of the mathematical models in terms of physiological meaning and to illustrate the accuracy and efficiency of the numerical methods.

Keywords: finite element method, cardiovascular system, Eulerian framework, haemodynamics, heart valve, cardiomyocyte, red blood cell

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2215 Heterologous Expression of a Clostridium thermocellum Proteins and Assembly of Cellulosomes 'in vitro' for Biotechnology Applications

Authors: Jessica Pinheiro Silva, Brenda Rabello De Camargo, Daniel Gusmao De Morais, Eliane Ferreira Noronha

Abstract:

The utilization of lignocellulosic biomass as source of polysaccharides for industrial applications requires an arsenal of enzymes with different mode of action able to hydrolyze its complex and recalcitrant structure. Clostridium thermocellum is gram-positive, thermophilic bacterium producing lignocellulosic hydrolyzing enzymes in the form of multi-enzyme complex, termed celulossomes. This complex has several hydrolytic enzymes attached to a large and enzymically inactive protein known as Cellulosome-integrating protein (CipA), which serves as a scaffolding protein for the complex produced. This attachment occurs through specific interactions between cohesin modules of CipA and dockerin modules in enzymes. The present work aims to construct celulosomes in vitro with the structural protein CipA, a xylanase called Xyn10D and a cellulose called CelJ from C.thermocellum. A mini-scafoldin was constructed from modules derived from CipA containing two cohesion modules. This was cloned and expressed in Escherichia coli. The other two genes were cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9 and integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. Purification of each protein is being carried out. Further studies regarding enzymatic activity of the cellulosome is going to be evaluated. The cellulosome built in vitro and composed of mini-CipA, CelJ and Xyn10D, can be very interesting for application in industrial processes involving the degradation of plant biomass.

Keywords: cellulosome, CipA, Clostridium thermocellum, cohesin, dockerin, yeast

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2214 Low Enrollment in Antiretroviral Treatment among Pregnant Women Screened HIV Infected in Informal Health Centers in Cameroon

Authors: Lydie Audrey Amboua Schouame, Sylvie Kwedi Nolna, Antoine Socpa, Alexandre Benjamin Nkoum

Abstract:

Background: Despite the struggle of the Cameroonian Ministry of Public Health against informal health centers (IHCs) because of their illegality, IHCs are booming in Cameroon and a large part of the population uses them. In 2017, more than 3.000 IHCs were counted across the country. Most of these IHCs have antenatal clinics and they screen pregnant women for HIV. However, there is no data on the Prevention of Mother-To-Child Transmission of HIV (PMTCT) in this informal health sector in Cameroon. This study aimed to investigate the initiation of Antiretroviral treatment (ART) in pregnant women screened HIV positive in IHCs and associated factors. Methods: From January 01, 2018, to June 30, 2020, we carried out a cohort study of pregnant women attending their first antenatal visit and screened HIV positive in informal health centers in the cities of Douala and Ebolowa in Cameroon. Consenting participants were interviewed at two points: at least one week after delivery of the HIV result and three months later. The collected data were entered into Kobo collected and analyzed in SPSS V23.0 software. Results: A total of 182 HIV-infected pregnant women were enrolled in the study. The median age at enrollment was 30 years (IQR, 24-34) and the median gestational age at first ANC was 25 weeks (IQR, 19-31). Overall 61% (111/182) had a secondary level of education, 65% (118/182) were married/in a common-law relationship and 69% (126/182) had no income activity. At their first ANC, 91% (166/182) were naïve to ARV treatment. Among them, only 45% (74/166) initiated ART. The median delay in initiating ARV treatment was 5 days (IQR, 0-25). Of those who have started ART, only 64% (48/74) remained on treatment 3 months later. Conclusion: In order to eliminate mother-to-child transmission of HIV, attention should be paid to IHCs.

Keywords: informal health centers, human immunodeficiency, antiretroviral treatment, pregnant women

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2213 Anti-Prostate Cancer Effect of GV-1001, a Novel Gonadotropin-Releasing Hormone Receptor Ligand

Authors: Ji Won Kim, Moo Yeol Lee, Keon Wook Kang

Abstract:

GV-1001, 16 amino acid fragment of human telomerase reverse transcriptase catalytic subunit (hTERT), has been developed as an injectable cancer vaccine for many types of solid tumors showing high-level of telomerase activity. In the present study, we evaluated the anti-cancer effect of GV-1001 on androgen-receptor-positive prostate cancer. Two signaling pathways, Gs-adenylate cyclase-cAMP and Gq-IP3-Ca2+ pathways play a central role in GnRH receptor (GnRHR)-mediated activities. We found that leuprolide acetate (LA) mainly acted on Gq-mediated Ca2+ signaling, while GV-1001 preferentially acted on cAMP signaling; and both the effects were counteracted by cetrorelix, a GnRHR antagonist. We further tested whether GV-1001 affects tumor growth of human prostate cancer cells in vivo. Prostate tumor xenografts were established using LNCap, androgen receptor-positive prostate cancer cells, and the nude mice bearing tumors were subcutaneously injected with GV-1001 (0.01, 0.1, 1, 10 microg/kg/day) and LA (0.01 microg/kg/day) for 2 weeks. GV-1001 (1 and 10 microg/kg/day) significantly inhibited tumor growth of LNCap xenografts. Interestingly, mRNA expression of MMP2 and MMP9 was significantly suppressed by GV-1001 injection, but not by LA administration. Boyden chamber assay revealed that GV-1001 potently inhibited cell migration of LNCap. Our finding suggests that GV-1001 as a novel GnRHR ligand, has anti-proliferative and anti-migratory effects on androgen receptor-positive prostate cancer cells.

Keywords: GV-1001, GnRH, hTERT, prostate cancer

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2212 LIZTOXD: Inclusive Lizard Toxin Database by Using MySQL Protocol

Authors: Iftikhar A. Tayubi, Tabrej Khan, Mansoor M. Alsubei, Fahad A. Alsaferi

Abstract:

LIZTOXD provides a single source of high-quality information about proteinaceous lizard toxins that will be an invaluable resource for pharmacologists, neuroscientists, toxicologists, medicinal chemists, ion channel scientists, clinicians, and structural biologists. We will provide an intuitive, well-organized and user-friendly web interface that allows users to explore the detail information of Lizard and toxin proteins. It includes common name, scientific name, entry id, entry name, protein name and length of the protein sequence. The utility of this database is that it can provide a user-friendly interface for users to retrieve the information about Lizard, toxin and toxin protein of different Lizard species. These interfaces created in this database will satisfy the demands of the scientific community by providing in-depth knowledge about Lizard and its toxin. In the next phase of our project we will adopt methodology and by using A MySQL and Hypertext Preprocessor (PHP) which and for designing Smart Draw. A database is a wonderful piece of equipment for storing large quantities of data efficiently. The users can thus navigate from one section to another, depending on the field of interest of the user. This database contains a wealth of information on species, toxins, toxins, clinical data etc. LIZTOXD resource that provides comprehensive information about protein toxins from lizard toxins. The combination of specific classification schemes and a rich user interface allows researchers to easily locate and view information on the sequence, structure, and biological activity of these toxins. This manually curated database will be a valuable resource for both basic researchers as well as those interested in potential pharmaceutical and agricultural applications of lizard toxins.

Keywords: LIZTOXD, MySQL, PHP, smart draw

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2211 Awareness on Department of Education’s Disaster Risk Reduction Management Program at Oriental Mindoro National High School: Basis for Support School DRRM Program

Authors: Nimrod Bantigue

Abstract:

The Department of Education is continuously providing safe teaching-learning facilities and hazard-free environments to the learners. To achieve this goal, teachers’ awareness of DepEd’s DRRM programs and activities is extremely important; thus, this descriptive correlational quantitative study was conceptualized. This research answered four questions on the profile and level of awareness of the 153 teacher respondents of Oriental Mindoro National High School for the academic year 2018-2019. Stratified proportional sampling was employed, and both descriptive and inferential statistics were utilized to treat data. The findings revealed that the majority of the teachers at OMNHS are female and are in the age bracket of 20-40. Most are married and pursue graduate studies. They have moderate awareness of the Department of Education’s DRRM programs and activities in terms of assessment of risks activities, planning activities, implementation activities during disaster and evaluation and monitoring activities with 3.32, 3.12, 3.40 and 3.31 as computed means, respectively. Further, the result showed a significant relationship between the profile of the respondents such as age, civil status and educational attainment and the level of awareness. On the contrary, sex does not have a significant relationship with the level of awareness. The Support School DRRM program with Utilization Guide on School DRRM Manual was proposed to increase, improve and strengthen the weakest areas of awareness rated in each DRRM activity, such as assessment of risks, planning, and implementation during disasters and monitoring and evaluation.

Keywords: awareness, management, monitoring, risk reduction

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2210 Parking Service Effectiveness at Commercial Malls

Authors: Ahmad AlAbdullah, Ali AlQallaf, Mahdi Hussain, Mohammed AlAttar, Salman Ashknani, Magdy Helal

Abstract:

We study the effectiveness of the parking service provided at Kuwaiti commercial malls and explore potential problems and feasible improvements. Commercial malls are important to Kuwaitis as the entertainment and shopping centers due to the lack of other alternatives. The difficulty and relatively long times wasted in finding a parking spot at the mall are real annoyances. We applied queuing analysis to one of the major malls that offer paid-parking (1040 parking spots) in addition to free parking. Patrons of the mall usually complained of the traffic jams and delays at entering the paid parking (average delay to park exceeds 15 min for about 62% of the patrons, while average time spent in the mall is about 2.6 hours). However, the analysis showed acceptable service levels at the check-in gates of the parking garage. Detailed review of the vehicle movement at the gateways indicated that arriving and departing cars both had to share parts of the gateway to the garage, which caused the traffic jams and delays. A simple comparison we made indicated that the largest commercial mall in Kuwait does not suffer such parking issues, while other smaller, yet important malls do, including the one we studied. It was suggested that well-designed inlets and outlets of that gigantic mall permitted smooth parking despite being totally free and mall is the first choice for most people for entertainment and shopping. A simulation model is being developed for further analysis and verification. Simulation can overcome the mathematical difficulty in using non-Poisson queuing models. The simulation model is used to explore potential changes to the parking garage entrance layout. And with the inclusion of the drivers’ behavior inside the parking, effectiveness indicators can be derived to address the economic feasibility of extending the parking capacity and increasing service levels. Outcomes of the study are planned to be generalized as appropriate to other commercial malls in Kuwait

Keywords: commercial malls, parking service, queuing analysis, simulation modeling

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2209 Detection, Isolation, and Raman Spectroscopic Characterization of Acute and Chronic Staphylococcus aureus Infection in an Endothelial Cell Culture Model

Authors: Astrid Tannert, Anuradha Ramoji, Christina Ebert, Frederike Gladigau, Lorena Tuchscherr, Jürgen Popp, Ute Neugebauer

Abstract:

Staphylococcus aureus is a facultative intracellular pathogen, which by entering host cells may evade immunologic host response as well as antimicrobial treatment. In that way, S. aureus can cause persistent intracellular infections which are difficult to treat. Depending on the strain, S. aureus may persist at different intracellular locations like the phagolysosome. The first barrier invading pathogens from the blood stream that they have to cross are the endothelial cells lining the inner surface of blood and lymphatic vessels. Upon proceeding from an acute to a chronic infection, intracellular pathogens undergo certain biochemical and structural changes including a deceleration of metabolic processes to adopt for long-term intracellular survival and the development of a special phenotype designated as small colony variant. In this study, the endothelial cell line Ea.hy 926 was used as a model for acute and chronic S. aureus infection. To this end, Ea.hy 926 cells were cultured on QIAscout™ Microraft Arrays, a special graded cell culture substrate that contains around 12,000 microrafts of 200 µm edge length. After attachment to the substrate, the endothelial cells were infected with GFP-expressing S. aureus for 3 weeks. The acute infection and the development of persistent bacteria was followed by confocal laser scanning microscopy, scanning the whole Microraft Array for the presence and for detailed determination of the intracellular location of fluorescent intracellular bacteria every second day. After three weeks of infection representative microrafts containing infected cells, cells with protruded infections and cells that did never show any infection were isolated and fixed for Raman micro-spectroscopic investigation. For comparison, also microrafts with acute infection were isolated. The acquired Raman spectra are correlated with the fluorescence microscopic images to give hints about a) the molecular alterations in endothelial cells during acute and chronic infection compared to non-infected cells, and b) metabolic and structural changes within the pathogen when entering a mode of persistence within host cells. We thank Dr. Ruth Kläver from QIAGEN GmbH for her support regarding QIAscout technology. Financial support by the BMBF via the CSCC (FKZ 01EO1502) and from the DFG via the Jena Biophotonic and Imaging Laboratory (JBIL, FKZ PO 633/29-1, BA 1601/10-1) is highly acknowledged.

Keywords: correlative image analysis, intracellular infection, pathogen-host adaption, Raman micro-spectroscopy

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2208 Using 3D Satellite Imagery to Generate a High Precision Canopy Height Model

Authors: M. Varin, A. M. Dubois, R. Gadbois-Langevin, B. Chalghaf

Abstract:

Good knowledge of the physical environment is essential for an integrated forest planning. This information enables better forecasting of operating costs, determination of cutting volumes, and preservation of ecologically sensitive areas. The use of satellite images in stereoscopic pairs gives the capacity to generate high precision 3D models, which are scale-adapted for harvesting operations. These models could represent an alternative to 3D LiDAR data, thanks to their advantageous cost of acquisition. The objective of the study was to assess the quality of stereo-derived canopy height models (CHM) in comparison to a traditional LiDAR CHM and ground tree-height samples. Two study sites harboring two different forest stand types (broadleaf and conifer) were analyzed using stereo pairs and tri-stereo images from the WorldView-3 satellite to calculate CHM. Acquisition of multispectral images from an Unmanned Aerial Vehicle (UAV) was also realized on a smaller part of the broadleaf study site. Different algorithms using two softwares (PCI Geomatica and Correlator3D) with various spatial resolutions and band selections were tested to select the 3D modeling technique, which offered the best performance when compared with LiDAR. In the conifer study site, the CHM produced with Corelator3D using only the 50-cm resolution panchromatic band was the one with the smallest Root-mean-square deviation (RMSE: 1.31 m). In the broadleaf study site, the tri-stereo model provided slightly better performance, with an RMSE of 1.2 m. The tri-stereo model was also compared to the UAV, which resulted in an RMSE of 1.3 m. At individual tree level, when ground samples were compared to satellite, lidar, and UAV CHM, RMSE were 2.8, 2.0, and 2.0 m, respectively. Advanced analysis was done for all of these cases, and it has been noted that RMSE is reduced when the canopy cover is higher when shadow and slopes are lower and when clouds are distant from the analyzed site.

Keywords: very high spatial resolution, satellite imagery, WorlView-3, canopy height models, CHM, LiDAR, unmanned aerial vehicle, UAV

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2207 Sensitivity of Acanthamoeba castellanii-Grown Francisella to Three Different Disinfectants

Authors: M. Knezevic, V. Marecic, M. Ozanic, I. Kelava, M. Mihelcic, M. Santic

Abstract:

Francisella tularensis is a highly infectious, gram-negative intracellular bacterium and the causative agent of tularemia. The bacterium has been isolated from more than 250 wild species, including protozoa cells. Since Francisella is very virulent and persists in the environment for years, the aim of this study was to investigate whether Acanthamoeba castellanii-grown F. novicida exhibits an alteration in the resistance to disinfectants. It has been shown by other intracellular pathogens, including Legionella pneumophila that bacteria grown in amoeba exhibit more resistance to disinfectants. However, there is no data showing Francisella viability behaviour after intracellular life cycle in A. castellani. In this study, the bacterial suspensions of A. castellanii-grown or in vitro-grown Francisella were treated with three different disinfectants, and the bacterial viability after disinfection treatment was determined by a colony-forming unit (CFU) counting method, transmission electron microscopy (TEM), fluorescence microscopy as well as the leakage of intracellular fluid. Our results have shown that didecyldimethylammonium chloride (DDAC) combined with isopropyl alcohol was the most effective in bacterial killing; all in vitro-grown and A. castellanii-grown F. novicida were killed after only 10s. Surprisingly, in comparison to in vitro-grown bacteria, A. castellanii-grown F. novicida was more sensitive to decontamination by the benzalkonium chloride combined with DDAC and formic acid and the polyhexamethylene biguanide (PHMB). We can conclude that the tested disinfectants exhibit antimicrobial activity by causing a loss of structural organization and integrity of the Francisella cell wall and membrane and the subsequent leakage of the intracellular contents. Finally, the results of this study clearly demonstrate that Francisella grown in A. castellanii had become more susceptible to many disinfectants.

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

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2206 Purification of Bacillus Lipopeptides for Diverse Applications

Authors: Vivek Rangarajan, Kim G. Clarke

Abstract:

Bacillus lipopeptides are biosurfactants with wide ranging applications in the medical, food, agricultural, environmental and cosmetic industries. They are produced as a mix of three families, surfactin, iturin and fengycin, each comprising a large number of homologues of varying functionalities. Consequently, the method and degree of purification of the lipopeptide cocktail becomes particularly important if the functionality of the lipopeptide end-product is to be maximized for the specific application. However, downstream processing of Bacillus lipopeptides is particularly challenging due to the subtle variations observed in the different lipopeptide homologues and isoforms. To date, the most frequently used lipopeptide purification operations have been acid precipitation, solvent extraction, membrane ultrafiltration, adsorption and size exclusion. RP-HPLC (reverse phase high pressure liquid chromatography) also has potential for fractionation of the lipopeptide homologues. In the studies presented here, membrane ultrafiltration and RP-HPLC were evaluated for lipopeptide purification to different degrees of purities for maximum functionality. Batch membrane ultrafiltration using 50 kDa polyether sulphone (PES) membranes resulted in lipopeptide recovery of about 68% for surfactin and 82 % for fengycin. The recovery was further improved to 95% by using size-conditioned lipopeptide micelles. The conditioning of lipopeptides with Ca2+ ions resulted in uniformly sized micelles with average size of 96.4 nm and a polydispersity index of 0.18. The size conditioning also facilitated removal of impurities (molecular weight ranging between 2335-3500 Da) through operation of the system under dia-filtration mode, in a way similar to salt removal from protein by dialysis. The resultant purified lipopeptide was devoid of macromolecular impurities and could ideally suit applications in the cosmetic and food industries. Enhanced purification using RP-HPLC was carried out in an analytical C18 column, with the aim to fractionate lipopeptides into their constituent homologues. The column was eluted with mobile phase comprising acetonitrile and water over an acetonitrile gradient, 35% - 80%, over 70 minutes. The gradient elution program resulted in as many as 41 fractions of individual lipopeptide homologues. The efficacy test of these fractions against fungal phytopathogens showed that first 21 fractions, identified to be homologues of iturins and fengycins, displayed maximum antifungal activities, suitable for biocontrol in the agricultural industry. Thus, in the current study, the downstream processing of lipopeptides leading to tailor-made products for selective applications was demonstrated using two major downstream unit operations.

Keywords: bacillus lipopeptides, membrane ultrafiltration, purification, RP-HPLC

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