Search results for: sequence spaces
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2294

Search results for: sequence spaces

524 Sustainable Agriculture Practices Using Bacterial-mediated Alleviation of Salinity Stress in Crop Plants

Authors: Mohamed Trigui, Fatma Masmoudi, Imen Zouari

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Massive utilizations of chemical fertilizer and chemical pesticides in agriculture sector to improve the farming productivity have created increasing environmental damages. Then, agriculture must become sustainable, focusing on production systems that respect the environment and help to reduce climate change. Isolation and microbial identification of new bacterial strains from naturally saline habitats and compost extracts could be a prominent way in pest management and crop production under saline conditions. In this study, potential mechanisms involved in plant growth promotion and suppressive activity against fungal diseases of a compost extract produced from poultry manure/olive husk compost and halotolerant and halophilic bacterial strains under saline stress were investigated. On the basis of the antimicrobial tests, different strains isolated from Sfax solar saltern (Tunisia) and from compost extracts were selected and tested for their plant growth promoting traits, such as siderophores production, nitrogen fixation, phosphate solubilization and the production of extracellular hydrolytic enzymes (protease and lipase) under in-vitro conditions. Among 450 isolated bacterial strains, 16 isolates showed potent antifungal activity against the tested plant pathogenic fungi. Their identification based on 16S rRNA gene sequence revealed they belonged to different species. Some of these strains were also characterized for their plant growth promoting capacities. Obtained results showed the ability of four strains belonging to Bacillus genesis to ameliorate germination rate and root elongation compared to the untreated positive controls. Combinatorial capacity of halotolerant bacteria with antimicrobial activity and plant growth promoting traits could be promising sources of interesting bioactive substances under saline stress.

Keywords: abiotic stress, biofertilizer, biotic stress, compost extract, halobacteria, plant growth promoting (PGP), soil fertility

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523 Monitoring Peri-Urban Growth and Land Use Dynamics with GIS and Remote Sensing Techniques: A Case Study of Burdwan City, India

Authors: Mohammad Arif, Soumen Chatterjee, Krishnendu Gupta

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The peri-urban interface is an area of transition where the urban and rural areas meet and interact. So the peri-urban areas, which is characterized by strong urban influence, easy access to markets, services and other inputs, are ready supplies of labour but distant from the land paucity and pollution related to urban growth. Hence, the present study is primarily aimed at quantifying the spatio-temporal pattern of land use/land cover change during the last three decades (i.e., 1987 to 2016) in the peri-urban area of Burdwan city. In the recent past, the morphology of the study region has rapid change due to high growth of population and establishment of industries. The change has predominantly taken place along the State and National Highway 2 (NH-2) and around the Burdwan Municipality for meeting both residential and commercial purposes. To ascertain the degree of change in land use and land cover, over the specified time, satellite imageries and topographical sheets are employed. The data is processed through appropriate software packages to arrive at a deduction that most of the land use changes have occurred by obliterating agricultural land & water bodies and substituting them by built area and industrial spaces. Geospatial analysis of study area showed that this area has experienced a steep increase (30%) of built-up areas and excessive decrease (15%) in croplands between 1987 and 2016. Increase in built-up areas is attributed to the increase of out-migration during this period from the core city. This study also examined social, economic and institutional factors that lead to this rapid land use change in peri-urban areas of the Burdwan city by carrying out a field survey of 250 households in peri-urban areas. The research concludes with an urgency for regulating land subdivisions in peri-urban areas to prevent haphazard land use development. It is expected that the findings of the study would go a long way in facilitating better policy making.

Keywords: growth, land use land cover, morphology, peri-urban, policy making

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522 A Genre-Based Approach to the Teaching of Pronunciation

Authors: Marden Silva, Danielle Guerra

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Some studies have indicated that pronunciation teaching hasn’t been paid enough attention by teachers regarding EFL contexts. In particular, segmental and suprasegmental features through genre-based approach may be an opportunity on how to integrate pronunciation into a more meaningful learning practice. Therefore, the aim of this project was to carry out a survey on some aspects related to English pronunciation that Brazilian students consider more difficult to learn, thus enabling the discussion of strategies that can facilitate the development of oral skills in English classes by integrating the teaching of phonetic-phonological aspects into the genre-based approach. Notions of intelligibility, fluency and accuracy were proposed by some authors as an ideal didactic sequence. According to their proposals, basic learners should be exposed to activities focused on the notion of intelligibility as well as intermediate students to the notion of fluency, and finally more advanced ones to accuracy practices. In order to test this hypothesis, data collection was conducted during three high school English classes at Federal Center for Technological Education of Minas Gerais (CEFET-MG), in Brazil, through questionnaires and didactic activities, which were recorded and transcribed for further analysis. The genre debate was chosen to facilitate the oral expression of the participants in a freer way, making them answering questions and giving their opinion about a previously selected topic. The findings indicated that basic students demonstrated more difficulty with aspects of English pronunciation than the others. Many of the intelligibility aspects analyzed had to be listened more than once for a better understanding. For intermediate students, the speeches recorded were considerably easier to understand, but nevertheless they found it more difficult to pronounce the words fluently, often interrupting their speech to think about what they were going to say and how they would talk. Lastly, more advanced learners seemed to express their ideas more fluently, but still subtle errors related to accuracy were perceptible in speech, thereby confirming the proposed hypothesis. It was also seen that using genre-based approach to promote oral communication in English classes might be a relevant method, considering the socio-communicative function inherent in the suggested approach.

Keywords: EFL, genre-based approach, oral skills, pronunciation

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521 Extracellular Production of the Oncolytic Enzyme, Glutaminase Free L-Asparaginase, from Newly Isolated Streptomyces Olivaceus NEAE-119: Optimization of Culture Conditions Using Response Surface Methodology

Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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520 Urban Health and Strategic City Planning: A Case from Greece

Authors: Alexandra P. Alexandropoulou, Andreas Fousteris, Eleni Didaskalou, Dimitrios A. Georgakellos

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As urbanization is becoming a major stress factor not only for the urban environment but also for the wellbeing of city dwellers, incorporating the issues of urban health in strategic city planning and policy-making has never been more relevant. The impact of urbanization can vary from low to severe and relates to all non-communicable diseases caused by the different functions of cities. Air pollution, noise pollution, water and soil pollution, availability of open green spaces, and urban heat island are the major factors that can compromise citizens' health. Urban health describes the effects of the social environment, the physical environment, and the availability and accessibility to health and social services. To assess the quality of urban wellbeing, all urban characteristics that might have an effect on citizens' health must be considered, evaluated, and introduced in integrated local planning. A series of indices and indicators can be used to better describe these effects and set the target values in policy making. Local strategic planning is one of the most valuable development tools a local city administration can possess; thus, it has become mandatory under Greek law for all municipalities. It involves a two-stage procedure; the first aims to collect, analyse and evaluate data on the current situation of the city (administrative data, population data, environmental data, social data, swot analysis), while the second aims to introduce a policy vision described and supported by distinct (nevertheless integrated) actions, plans and measures to be implemented with the aim of city development and citizen wellbeing. In this procedure, the element of health is often neglected or under-evaluated. A relative survey was conducted among all Greek local authorities in order to shed light on the current situation. Evidence shows that the rate of incorporation of health in strategic planning is lacking behind. The survey also highlights key hindrances and concerns raised by local officials and suggests a path for the way forward.

Keywords: urban health, strategic planning, local authorities, integrated development

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519 Transformation of Traditional Marketplaces in an Urban Context: Case of Chalai Market, Thiruvananthapuram

Authors: Aswathy Vijayan, Sharath Sunder Rajeev

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Trade has been fundamental in the footprint of human civilization since ancient time. In most of the historic cities, city development was along trading routes, where marketplaces are the major entrance to a city and hence a major element of the urban fabric. Marketplaces are where the commercial activities flourish, people, having a sense of belonging to the place, where they easily fit in. Acknowledging the built environment in and around the market in a way, creating a sense of place is an important factor in the success of public spaces. Local markets are developed in an organic manner, which adds on to the people experience and perception of urban space. With the city development, the commercial needs within the city increase, hence marketplaces flourish, irrespective of the functional segregation within. The work-live culture in the marketplaces diminishes as the commercial expansion washes away the residential patches within it. Real estate flourishes as the newer infills are without considering the carrying capacity of the place. Chalai market is a prominent business center serving the regional level of Thiruvananthapuram city. The transformation trend of marketplaces in city cores are understood from case study on Fatimid Cairo Marketplace. The parameters that led to transformation of marketplaces in a global context is considered for the analysis of the Chalai market. The structure of the marketplace over the years is analyzed in terms of transformation in location, transformation in the land- use, change in commodity, and transformation in movement and activity. The aim of the research is to emphasize the need to understand the transformation trend, in creating a suitable development pattern for the city. The unregulated transformation within the city core has led to tremendous transformation in the user group and user pattern and eventually to the commercial trend. With the change in lifestyle and need for new amenities have led to addition of new infills leading to the degradation of the native commerce. Hence addressing the transformation of marketplaces are crucial to maintaining the locational significance and cultural importance and heritage of the place.

Keywords: bazaar, market centers, marketplaces, traditional city, traditional market, urban fabric

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518 Collaborative Approaches in Achieving Sustainable Private-Public Transportation Services in Inner-City Areas: A Case of Durban Minibus Taxis

Authors: Lonna Mabandla, Godfrey Musvoto

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Transportation is a catalytic feature in cities. Transport and land use activity are interdependent and have a feedback loop between how land is developed and how transportation systems are designed and used. This recursive relationship between land use and transportation is reflected in how public transportation routes internal to the inner-city enhance accessibility, therefore creating spaces that are conducive to business activity, while the business activity also informs public transportation routes. It is for this reason that the focus of this research is on public transportation within inner-city areas where the dynamic is evident. Durban is the chosen case study where the dominating form of public transportation within the central business district (CBD) is minibus taxis. The paradox here is that minibus taxis still form part of the informal economy even though they are the leading form of public transportation in South Africa. There have been many attempts to formalise this industry to follow more regulatory practices, but minibus taxis are privately owned, therefore complicating any proposed intervention. The argument of this study is that the application of collaborative planning through a sustainable partnership between the public and private sectors will improve the social and environmental sustainability of public transportation. One of the major challenges that exist within such collaborative endeavors is power dynamics. As a result, a key focus of the study is on power relations. Practically, power relations should be observed over an extended period, specifically when the different stakeholders engage with each other, to reflect valid data. However, a lengthy data collection process was not possible to observe during the data collection phase of this research. Instead, interviews were conducted focusing on existing procedural planning practices between the inner-city minibus taxi association (South and North Beach Taxi Association), the eThekwini Transport Authority (ETA), and the eThekwini Town Planning Department. Conclusions and recommendations were then generated based on these data.

Keywords: collaborative planning, sustainability, public transport, minibus taxis

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517 Hidden Hot Spots: Identifying and Understanding the Spatial Distribution of Crime

Authors: Lauren C. Porter, Andrew Curtis, Eric Jefferis, Susanne Mitchell

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A wealth of research has been generated examining the variation in crime across neighborhoods. However, there is also a striking degree of crime concentration within neighborhoods. A number of studies show that a small percentage of street segments, intersections, or addresses account for a large portion of crime. Not surprisingly, a focus on these crime hot spots can be an effective strategy for reducing community level crime and related ills, such as health problems. However, research is also limited in an important respect. Studies tend to use official data to identify hot spots, such as 911 calls or calls for service. While the use of call data may be more representative of the actual level and distribution of crime than some other official measures (e.g. arrest data), call data still suffer from the 'dark figure of crime.' That is, there is most certainly a degree of error between crimes that occur versus crimes that are reported to the police. In this study, we present an alternative method of identifying crime hot spots, that does not rely on official data. In doing so, we highlight the potential utility of neighborhood-insiders to identify and understand crime dynamics within geographic spaces. Specifically, we use spatial video and geo-narratives to record the crime insights of 36 police, ex-offenders, and residents of a high crime neighborhood in northeast Ohio. Spatial mentions of crime are mapped to identify participant-identified hot spots, and these are juxtaposed with calls for service (CFS) data. While there are bound to be differences between these two sources of data, we find that one location, in particular, a corner store, emerges as a hot spot for all three groups of participants. Yet it does not emerge when we examine CFS data. A closer examination of the space around this corner store and a qualitative analysis of narrative data reveal important clues as to why this store may indeed be a hot spot, but not generate disproportionate calls to the police. In short, our results suggest that researchers who rely solely on official data to study crime hot spots may risk missing some of the most dangerous places.

Keywords: crime, narrative, video, neighborhood

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516 Rheumatoid Arthritis, Periodontitis and the Subgingival Microbiome: A Circular Relationship

Authors: Isabel Lopez-Oliva, Akshay Paropkari, Shweta Saraswat, Stefan Serban, Paola de Pablo, Karim Raza, Andrew Filer, Iain Chapple, Thomas Dietrich, Melissa Grant, Purnima Kumar

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Objective: We aimed to explicate the role of the subgingival microbiome in the causal link between rheumatoid arthritis (RA) and periodontitis (PD). Methods: Subjects with/without RA and with/without PD were randomized for treatment with scaling and root planing (SRP) or oral hygiene instructions. Subgingival biofilm, gingival crevicular fluid, and serum were collected at baseline and at 3- and 6-months post-operatively. Correlations were generated between 72 million 16S rDNA sequences, immuno-inflammatory mediators, circulating antibodies to oral microbial antigens, serum inflammatory molecules, and clinical metrics of RA. The dynamics of inter-microbial and host-microbial interactions were modeled using differential network analysis. Results: RA superseded periodontitis as a determinant of microbial composition, and DAS28 score superseded the severity of periodontitis as a driver of microbial assemblages (p=0.001, ANOSIM). RA subjects evidenced higher serum anti-PPAD (p=0.0013), anti-Pg-enolase (p=0.0031), anti-RPP3, anti- Pg-OMP and anti- Pi-OMP (p=0.001) antibodies than non-RA controls (with and without periodontitis). Following SRP, bacterial networks anchored by IL-1b, IL-4, IL-6, IL-10, IL-13, MIP-1b, and PDGF-b underwent ≥5-fold higher rewiring; and serum antibodies to microbial antigens decreased significantly. Conclusions: Our data suggest a circular relationship between RA and PD, beginning with an RA-influenced dysbiosis within the healthy subgingival microbiome that leads to exaggerated local inflammation in periodontitis and circulating antibodies to periodontal pathogens and positive correlation between severity of periodontitis and RA activity. Periodontal therapy restores host-microbial homeostasis, reduces local inflammation, and decreases circulating microbial antigens. Our data highlights the importance of integrating periodontal care into the management of RA patients.

Keywords: rheumatoid arthritis, periodontal, subgingival, DNA sequence analysis, oral microbiome

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515 Development of Knitted Seersucker Fabric for Improved Comfort Properties

Authors: Waqas Ashraf, Yasir Nawab, Haritham Khan, Habib Awais, Shahbaz Ahmad

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Seersucker is a popular lightweight fabric widely used in men’s and women’s suiting, casual wear, children’s clothing, house robes, bed spreads and for spring and summer wear. The puckered effect generates air spaces between body and the fabric, keeping the wearer cool in hot conditions. The aim of this work was to develop knitted seersucker fabric on single cylinder weft knitting machine using plain jersey structure. Core spun cotton yarn and cotton spun yarn of same linear density were used. Core spun cotton yarn, contains cotton fiber in the sheath and elastase filament in the core. The both yarn were fed at regular interval to feeders on the machine. The loop length and yarn tension were kept constant at each feeder. The samples were then scoured and bleached. After wet processing, the fabric samples were washed and tumble dried. Parameters like loop length, stitch density and areal density were measured after conditioning these samples for 24 hours in Standard atmospheric condition. Produced sample has a regular puckering stripe along the width of the fabric with same height. The stitch density of both the flat and puckered area of relaxed fabric was found to be different .Air permeability and moisture management tests were performed. The results indicated that the knitted seersucker fabric has better wicking and moisture management properties as the flat area contact, whereas puckered area held away from the skin. Seersucker effect in knitted fabric was achieved by the difference of contraction of both sets of courses produced from different types of yarns. The seer sucker fabric produce by knitting technique is less expensive as compared to woven seer sucker fabric as there is no need of yarn preparation. The knitted seersucker fabric is more practicable for summer dresses, skirts, blouses, shirts, trousers and shorts.

Keywords: air permeability, knitted structure, moisture management, seersucker

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514 Role of Matric Suction in Mechanics behind Swelling Characteristics of Expansive Soils

Authors: Saloni Pandya, Nikhil Sharma, Ajanta Sachan

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Expansive soils in the unsaturated state are part of vadose zone and encountered in several arid and semi-arid parts of the world. Influence of high temperature, low precipitation and alternate cycles of wetting and drying are responsible for the chemical weathering of rocks, which results in the formation of expansive soils. Shrinkage-swelling (expansive) soils cover a substantial portion of area in India. Damages caused by expansive soils to various geotechnical structures are alarming. Matric suction develops in unsaturated soil due to capillarity and surface tension phenomena. Matric suction influences the geometric arrangement of soil skeleton, which induces the volume change behaviour of expansive soil. In the present study, an attempt has been made to evaluate the role of matric suction in the mechanism behind swelling characteristics of expansive soil. Four different soils have been collected from different parts of India for the current research. Soil sample S1, S2, S3 and S4 were collected from Nagpur, Bharuch, Bharuch-Dahej highway and Ahmedabad respectively. DFSI (Differential Free Swell Index) of these soils samples; S1, S2, S3, and S4; were determined to be 134%, 104%, 70% and 30% respectively. X-ray diffraction analysis of samples exhibited that percentage of Montmorillonite mineral present in the soils reduced with the decrease in DFSI. A series of constant volume swell pressure tests and in-contact filter paper tests were performed to evaluate swelling pressure and matric suction of all four soils at 30% saturation and 1.46 g/cc dry density. Results indicated that soils possessing higher DFSI exhibited higher matric suction as compared to lower DFSI expansive soils. Significant influence of matric suction on swelling pressure of expansive soils was observed with varying DFSI values. Higher matric suction of soil might govern the water uptake in the interlayer spaces of Montmorillonite mineral present in expansive soil leading to crystalline swelling.

Keywords: differential free swell index, expansive soils, matric suction, swelling pressure

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513 Synergistic Anti-Proliferation Effect of PLK-1 Inhibitor and Livistona Chinensis Fruit Extracts on Lung Adenocarcinoma A549 Cells

Authors: Min-Chien Su, Tzu-Hsuan Hsu, Guan-Xuan Wu, Shyh-Ming Kuo

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Lung cancer is one of the clinically challenging malignant diseases worldwide. For efficient therapeutics in cancer, combination therapy has developed to acquire a better outcome. PLK-1 was one of the major factors affecting cell mitosis in cancer cells, its inhibitor Bi6727 was proven effective in treating several different cancers namely oral cancer, colon cancer and lung cancer. Despite its low toxicity toward normal cells compared to traditional chemotherapy, it is still yet to be evaluated in detail. Livistona Chinensis (LC) is a Chinese herb that used as a traditional prescription to treat lung cancer. Due to the uncertainty of the efficacy of LC, we utilized a water extraction method to extract the Livistona Chinensis and then lyophilized into powder for further study. In this study we investigated the antiproliferation activities of Bi6727 and LC extracts (LCE) on A549 non-small lung cancer cells. The IC50 of Bi6727 and LCE on A549 are 60 nM and 0.8 mg/mL, respectively. The fluorescent staining images shown nucleolus damage in cells treated with Bi6727 and mitochondrial damage after treated with LCE. A549 cells treated with Bi6727 and LCE showed increased expression of Bax, Caspase-3 and Caspase-9 proteins from Western blot assay. LCE also inhibited A549 cells growth keeping cells at G2-M phase from cell cycle assay. Apoptosis assay results showed that LCE induced late apoptosis of A549 cells. JC-1 assay showed that the mitochondria damaged at the LCE concentration of 0.4 mg/mL. In our preliminary anti-proliferation test of combined LCE and Bi-6727 on A549 cells, we found a dramatically decrease in proliferation after treated with LCE first for 24-h and then Bi-6727 for extra 24-h. This was an important finding regarding synergistic anti-proliferation effect of these drugs, However, the usage, the application sequence of LCE and Bi-6727 on A549 cells and their related mechanisms still need to be evaluated. In summary, the drugs exerted anti-proliferation effect on A549 cells independently. We hopefully combine the usage of these two drugs will bring a different and potential outcome in treating lung cancer.

Keywords: anti-proliferation, A549, Livistona Chinensis fruit extracts, PLK-1 inhibitor

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512 Epigenetic Modifying Potential of Dietary Spices: Link to Cure Complex Diseases

Authors: Jeena Gupta

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In the today’s world of pharmaceutical products, one should not forget the healing properties of inexpensive food materials especially spices. They are known to possess hidden pharmaceutical ingredients, imparting them the qualities of being anti-microbial, anti-oxidant, anti-inflammatory and anti-carcinogenic. Further aberrant epigenetic regulatory mechanisms like DNA methylation, histone modifications or altered microRNA expression patterns, which regulates gene expression without changing DNA sequence, contribute significantly in the development of various diseases. Changing lifestyles and diets exert their effect by influencing these epigenetic mechanisms which are thus the target of dietary phytochemicals. Bioactive components of plants have been in use since ages but their potential to reverse epigenetic alterations and prevention against diseases is yet to be explored. Spices being rich repositories of many bioactive constituents are responsible for providing them unique aroma and taste. Some spices like curcuma and garlic have been well evaluated for their epigenetic regulatory potential, but for others, it is largely unknown. We have evaluated the biological activity of phyto-active components of Fennel, Cardamom and Fenugreek by in silico molecular modeling, in vitro and in vivo studies. Ligand-based similarity studies were conducted to identify structurally similar compounds to understand their biological phenomenon. The database searching has been done by using Fenchone from fennel, Sabinene from cardamom and protodioscin from fenugreek as a query molecule in the different small molecule databases. Moreover, the results of the database searching exhibited that these compounds are having potential binding with the different targets found in the Protein Data Bank. Further in addition to being epigenetic modifiers, in vitro study had demonstrated the antimicrobial, antifungal, antioxidant and cytotoxicity protective effects of Fenchone, Sabinene and Protodioscin. To best of our knowledge, such type of studies facilitate the target fishing as well as making the roadmap in drug design and discovery process for identification of novel therapeutics.

Keywords: epigenetics, spices, phytochemicals, fenchone

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511 Using Geographic Information System and Analytic Hierarchy Process for Detecting Forest Degradation in Benslimane Forest, Morocco

Authors: Loubna Khalile, Hicham Lahlaoi, Hassan Rhinane, A. Kaoukaya, S. Fal

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Green spaces is an essential element, they contribute to improving the quality of lives of the towns around them. They are a place of relaxation, walk and rest a playground for sport and youths. According to United Nations Organization Forests cover 31% of the land. In Morocco in 2013 that cover 12.65 % of the total land area, still, a small proportion compared to the natural needs of forests as a green lung of our planet. The Benslimane Forest is a large green area It belongs to Chaouia-Ouardigha Region and Greater Casablanca Region, it is located geographically between Casablanca is considered the economic and business Capital of Morocco and Rabat the national political capital, with an area of 12261.80 Hectares. The essential problem usually encountered in suburban forests, is visitation and tourism pressure it is anthropogenic actions, as well as other ecological and environmental factors. In recent decades, Morocco has experienced a drought year that has influenced the forest with increasing human pressure and every day it suffers heavy losses, as well as over-exploitation. The Moroccan forest ecosystems are weak with intense ecological variation, domanial and imposed usage rights granted to the population; forests are experiencing a significant deterioration due to forgetfulness and immoderate use of forest resources which can influence the destruction of animal habitats, vegetation, water cycle and climate. The purpose of this study is to make a model of the degree of degradation of the forest and know the causes for prevention by using remote sensing and geographic information systems by introducing climate and ancillary data. Analytic hierarchy process was used to find out the degree of influence and the weight of each parameter, in this case, it is found that anthropogenic activities have a fairly significant impact has thus influenced the climate.

Keywords: analytic hierarchy process, degradation, forest, geographic information system

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510 Loss of Function of Only One of Two CPR5 Paralogs Causes Resistance Against Rice Yellow Mottle Virus

Authors: Yugander Arra, Florence Auguy, Melissa Stiebner, Sophie Chéron, Michael M. Wudick, Van Schepler-Luu, Sébastien Cunnac, Wolf B. Frommer, Laurence Albar

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Rice yellow mottle virus (RYMV) is one of the most important diseases affecting rice in Africa. The most promising strategy to reduce yield losses is the use of highly resistant varieties. The resistance gene RYMV2 is homolog of the Arabidopsis constitutive expression of pathogenesis related protein-5 (AtCPR5) nucleoporin gene. Resistance alleles are originating from African cultivated rice Oryza glaberrima, rarely cultivated, and are characterized by frameshifts or early stop codons, leading to a non-functional or truncated protein. Rice possesses two paralogs of CPR5 and function of these genes are unclear. Here, we evaluated the role of the two rice candidate nucleoporin paralogs OsCPR5.1 (pathogenesis-related gene 5; RYMV2) and OsCPR5.2 by CRISPR/Cas9 genome editing. Despite striking sequence and structural similarity, only loss-of-function of OsCPR5.1 led to full resistance, while loss-of-function oscpr5.2 mutants remained susceptible. Short N-terminal deletions in OsCPR5.1 also did not lead to resistance. In contrast to Atcpr5 mutants, neither OsCPR5.1 nor OsCPR5.2 knock out mutants showed substantial growth defects. Taken together, the candidate nucleoporin OsCPR5.1, but not its close homolog OsCPR5.2, plays a specific role for the susceptibility to RYMV, possibly by impairing the import of viral RNA or protein into the nucleus. Whereas gene introgression from O. glaberrima to high yielding O. sativa varieties is impaired by strong sterility barriers and the negative impact of linkage drag, genome editing of OsCPR5.1, while maintaining OsCPR5.2 activity, thus provides a promising strategy to generate O. sativa elite lines that are resistant to RYMV.

Keywords: CRISPR Cas9, genome editing, knock out mutant, recessive resistance, rice yellow mottle virus

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509 From Cultural Policy to Social Practice: Literary Festivals as a Platform for Social Inclusion in Pakistan

Authors: S. Jabeen

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Though Pakistan has a rich cultural history and a diverse population; its global image is tarnished with labels of Muslim ‘fundamentalism’ and ‘extremism.’ Cultural policy is a tool that can be used by the government of Pakistan to ameliorate this image, but instead, this fundamentalist reputation is reinforced in the 2005 draft of Pakistan’s cultural policy. With its stern focus on a homogenized cultural identity, this 2005 draft bases itself largely on forced participation from the largely Muslim public and leaves little or no benefits to them or cultural minorities in Pakistan. The effects of this homogenized ‘Muslim’ identity linger ten years later where the study and celebration of the cultural heritage of Pakistan in schools and educational festivals focus entirely on creating and maintaining a singular ‘Islamic’ cultural identity. The current lack of inclusion has many adverse effects that include the breeding of extremist mindsets through the usurpation of minority rights and lack of safe cultural public spaces. This paper argues that Pakistan can improve social inclusivity and boost its global image through cultural policy. The paper sets the grounds for research by surveying the effectiveness of different cultural policies across nations with differing socioeconomic status. Then, by sampling two public literary festivals in Pakistan as case studies, the National Youth Peace Festival hosted with a nationalistic agenda using public funds and the Lahore Literary Festival (LLF) that aims to boost the cultural literacy scene of Lahore using both private and public efforts, this paper looks at the success of the private, more inclusive LLF. A revision of cultural policy is suggested that combines public and private efforts to host cultural festivals for the sake of cultural celebration and human development, without a set nationalistic agenda. Consequently, this comparison which is grounded in the human capabilities approach, recommends revising the 2005 draft of the Cultural Policy to improve human capabilities in order to support cultural diversity and ultimately contribute to economic growth in Pakistan.

Keywords: cultural policy, festivals, human capabilities, Pakistan

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508 Urban Sustainable Development with Flood Crisis Management Approach

Authors: Ali Liaghat, Navid Tavanpour, Nima Tavanpour

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An increase in population and prevalence of urbanity have led plan makers and decision makers put effort into sustainable development of cities at national and local levels. One of the important issues in urban development is compliance with safety issues in cities. Despite natural disasters and unexpected events such as floods, earthquakes, hurricanes, fires, etc., urban development should be regarded as an axiom, or else any form of construction and development is not safe, because it will greatly harm economic growth and development and pose an obstacle to achieving sustainable development, plus a loss to lives and finances of people. Therefore, in line with urban development, it is necessary to identify particular environmental and local issues as determinants and pay attention to them at the top of everything, in that we can call it a good action and factor in urban sustainable developments. Physical structure of each city represents how it has developed or its development shaped and what incidents, changes, natural disasters it has undergone over time. Since any form of development plan should be in accordance with the previous situations of cities, disregarding it, unfortunately, can escalate into uncontrolled urban development, non-resistant and unstable construction against earthquake or invasion of river areas, destruction of agricultural lands or vegetation, periodic floods over time. It has been viewed as serious threats to developing cities, and typically caused destruction of bed and other urban facilities as well as damages to lives and finances. In addition, uncontrolled development has caused cities to look ugly in terms of urban façade, and off and on such unplanned measures caused the country to face countless losses, and it not only vitiates expenses incurred, but it will also impose additional costs of reconstruction, i.e. it is unsustainable development. Thus, in this paper, in addition to a discussion about necessity for a profound attitude toward this subject and making long-term plans, programs for organizing river and its surrounding area, creating open and green urban spaces, retrofitting and flood preventing are presented for sustainable safety and development of cities along with a critique of successful countries.

Keywords: flood, sustainable development, urbanisation, urban management

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507 Urban Renewal, Social Housing, Relocation, and Violence in Algiers

Authors: Kahina Amal Djiar, Mouna Gharbi, Maha Messaoudene, Oumelkheir Chareb

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Over the last decade, Algerian authorities have implemented an ambitious program of urban renewal, which includes important relocation operations. The objectives behind such strategic interventions are on the one hand, to carry out an incremental approach aiming at eradicating precarious housing and on the other hand, to diversify alternative housing options for families requiring better living spaces. It is precisely for these same purposes that the Djenan el-Hassan and Carrières Jaubert estates, which are both located in Algiers, have undergone major urban transformations. These dwelling sites were built as part of the famous "Battle of Housing", which was launched by French colonial administration in the 1950s just before the independence of Algeria in 1962. Today, the Djenan el-Hassan estate is almost entirely demolished following the relocation of 171 families. The Carrières Jaubert estate, for its part, has seen two kinds of operations. The first has been shaped by a process of urban requalification and redevelopment, which allowed some of the residents to stay on site after the transformation of most housing cells into larger apartments. The second operation has required the relocation of over 300 families to entirely newly built dwellings. Such projects of urban renewal are supposed to create new opportunities, not only in terms of local urban development, but also in terms of social perspectives for those families who are involved, either directly or indirectly, in the process of relocation. In fact, the percentage of urban violence in Algiers has increased instead. Recent events in the newly built estates show that residents are repeatedly experiencing and even instigating episodes of brutality, hostility and aggression. The objective of this paper is to examine the causes that have engendered such rise in urban violence in newly built housing estates in Algiers. This paper aims to present the findings of a recent qualitative research and highlight the way that poorly designed neighbourhood, combined with a relocation process that leaves little room for community participation, create inevitably severe social tensions.

Keywords: relocation, social housing, violence, Algiers

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506 Surface Water Flow of Urban Areas and Sustainable Urban Planning

Authors: Sheetal Sharma

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Urban planning is associated with land transformation from natural areas to modified and developed ones which leads to modification of natural environment. The basic knowledge of relationship between both should be ascertained before proceeding for the development of natural areas. Changes on land surface due to build up pavements, roads and similar land cover, affect surface water flow. There is a gap between urban planning and basic knowledge of hydrological processes which should be known to the planners. The paper aims to identify these variations in surface flow due to urbanization for a temporal scale of 40 years using Storm Water Management Mode (SWMM) and again correlating these findings with the urban planning guidelines in study area along with geological background to find out the suitable combinations of land cover, soil and guidelines. For the purpose of identifying the changes in surface flows, 19 catchments were identified with different geology and growth in 40 years facing different ground water levels fluctuations. The increasing built up, varying surface runoff are studied using Arc GIS and SWMM modeling, regression analysis for runoff. Resulting runoff for various land covers and soil groups with varying built up conditions were observed. The modeling procedures also included observations for varying precipitation and constant built up in all catchments. All these observations were combined for individual catchment and single regression curve was obtained for runoff. Thus, it was observed that alluvial with suitable land cover was better for infiltration and least generation of runoff but excess built up could not be sustained on alluvial soil. Similarly, basalt had least recharge and most runoff demanding maximum vegetation over it. Sandstone resulted in good recharging if planned with more open spaces and natural soils with intermittent vegetation. Hence, these observations made a keystone base for planners while planning various land uses on different soils. This paper contributes and provides a solution to basic knowledge gap, which urban planners face during development of natural surfaces.

Keywords: runoff, built up, roughness, recharge, temporal changes

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505 Wicking Bed Cultivation System as a Strategic Proposal for the Cultivation of Milpa and Mexican Medicinal Plants in Urban Spaces

Authors: David Lynch Steinicke, Citlali Aguilera Lira, Andrea León García

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The proposal posed in this work comes from a researching-action approach. In Mexico, a dialogue of knowledge may function as a link between traditional, local, pragmatic knowledge, and technological, scientific knowledge. The advantage of generating this nexus lies on the positive impact in the environment, in society and economy. This work attempts to combine, on the one hand the traditional Mexican knowledge such as the usage of medicinal herb and the agroecosystem milpa; and on the other hand make use of a newly created agricultural ecotechnology which main function is to take advantage of the urban space and to save water. This ecotechnology is the wicking bed. In a globalized world, is relevant to have a proposal where the most important aspect is to revalorize the culture through the acquisition of traditional knowledge but at the same time adapting them to the new social and urbanized structures without threatening the environment. The methodology used in this work comes from a researching-action approach combined with a practical dimension where an experimental model made of three wickingbeds was implemented. In this model, there were cultivated medicinal herb and milpa components. The water efficiency and the social acceptance were compared with a traditional ground crop, all this practice was made in an urban social context. The implementation of agricultural ecotechnology has had great social acceptance as its irrigation involves minimal effort and it is economically feasible for low-income people. The wicking bed system raised in this project is attainable to be implemented in schools, urban and peri-urban environments, homemade gardens and public areas. The proposal managed to carry out an innovative and sustainable knowledge-based traditional Mexican agricultural technology, allowing regain Milpa agroecosystem in urban environments to strengthen food security in favour of nutritional and protein benefits for the Mexican fare.

Keywords: milpa, traditional medicine, urban agriculture, wicking bed

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504 Conjunctive Management of Surface and Groundwater Resources under Uncertainty: A Retrospective Optimization Approach

Authors: Julius M. Ndambuki, Gislar E. Kifanyi, Samuel N. Odai, Charles Gyamfi

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Conjunctive management of surface and groundwater resources is a challenging task due to the spatial and temporal variability nature of hydrology as well as hydrogeology of the water storage systems. Surface water-groundwater hydrogeology is highly uncertain; thus it is imperative that this uncertainty is explicitly accounted for, when managing water resources. Various methodologies have been developed and applied by researchers in an attempt to account for the uncertainty. For example, simulation-optimization models are often used for conjunctive water resources management. However, direct application of such an approach in which all realizations are considered at each iteration of the optimization process leads to a very expensive optimization in terms of computational time, particularly when the number of realizations is large. The aim of this paper, therefore, is to introduce and apply an efficient approach referred to as Retrospective Optimization Approximation (ROA) that can be used for optimizing conjunctive use of surface water and groundwater over a multiple hydrogeological model simulations. This work is based on stochastic simulation-optimization framework using a recently emerged technique of sample average approximation (SAA) which is a sampling based method implemented within the Retrospective Optimization Approximation (ROA) approach. The ROA approach solves and evaluates a sequence of generated optimization sub-problems in an increasing number of realizations (sample size). Response matrix technique was used for linking simulation model with optimization procedure. The k-means clustering sampling technique was used to map the realizations. The methodology is demonstrated through the application to a hypothetical example. In the example, the optimization sub-problems generated were solved and analysed using “Active-Set” core optimizer implemented under MATLAB 2014a environment. Through k-means clustering sampling technique, the ROA – Active Set procedure was able to arrive at a (nearly) converged maximum expected total optimal conjunctive water use withdrawal rate within a relatively few number of iterations (6 to 7 iterations). Results indicate that the ROA approach is a promising technique for optimizing conjunctive water use of surface water and groundwater withdrawal rates under hydrogeological uncertainty.

Keywords: conjunctive water management, retrospective optimization approximation approach, sample average approximation, uncertainty

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503 Worldbuilding as Critical Architectural Pedagogy

Authors: Jesse Rafeiro

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This paper discusses worldbuilding as a pedagogical approach to the first-year architectural design studio. The studio ran for three consecutive terms between 2016-2018. Taking its departure from the fifty-five city narratives in Italo Calvino’s Invisible Cities, students collectively designed in a “nowhere” space where intersecting and diverging narratives could be played out. Along with Calvino, students navigated between three main exercises and their imposed limits to develop architectural insight at three scales simulating the considerations of architectural practice: detail, building, and city. The first exercise asked each student to design and model a ruin based on randomly assigned incongruent fragments. Each student was given one plan fragment and two section fragments from different Renaissance Treatises. The students were asked to translate these in alternating axonometric projection and model-making explorations. Although the fragments themselves were imposed, students were free to interpret how the drawings fit together by imagining new details and atypical placements. An undulating terrain model was introduced in the second exercise to ground the worldbuilding exercises. Here, students were required to negotiate with one another to design a city of ruins. Free to place their models anywhere on the site, the students were restricted by the negotiation of territories marked by other students and the requirement to provide thresholds, open spaces, and corridors. The third exercise introduced new life into the ruined city through a series of design interventions. Each student was assigned an atypical building program suggesting a place for an activity, human or nonhuman. The atypical nature of the programs challenged the triviality of functional planning through explorations in spatial narratives free from preconceived assumptions. By contesting, playing out, or dreaming responses to realities taught in other coursework, this third exercise actualized learnings that are too often self-contained in the silos of differing course agendas. As such, the studio fostered an initial worldbuilding space within which to sharpen sensibility and criticality for subsequent years of education.

Keywords: architectural pedagogy, critical pedagogy, Italo Calvino, worldbuilding

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502 Molecular Diversity of Forensically Relevant Insects from the Cadavers of Lahore

Authors: Sundus Mona, Atif Adnan, Babar Ali, Fareeha Arshad, Allah Rakha

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Molecular diversity is the variation in the abundance of species. Forensic entomology is a neglected field in Pakistan. Insects collected from the crime scene should be handled by forensic entomologists who are currently virtually non-existent in Pakistan. Correct identification of insect specimen along with knowledge of their biodiversity can aid in solving many problems related to complicated forensic cases. Inadequate morphological identification and insufficient thermal biological studies limit the entomological utility in Forensic Medicine. Recently molecular identification of entomological evidence has gained attention globally. DNA barcoding is the latest and established method for species identification. Only proper identification can provide a precise estimation of postmortem intervals. Arthropods are known to be the first tourists scavenging on decomposing dead matter. The objective of the proposed study was to identify species by molecular techniques and analyze their phylogenetic importance with barcoded necrophagous insect species of early succession on human cadavers. Based upon this identification, the study outcomes will be the utilization of established DNA bar codes to identify carrion feeding insect species for concordant estimation of post mortem interval. A molecular identification method involving sequencing of a 658bp ‘barcode’ fragment of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene from collected specimens of unknown dipteral species from cadavers of Lahore was evaluated. Nucleotide sequence divergences were calculated using MEGA 7 and Arlequin, and a neighbor-joining phylogenetic tree was generated. Three species were identified, Chrysomya megacephala, Chrysomya saffranea, and Chrysomya rufifacies with low genetic diversity. The fixation index was 0.83992 that suggests a need for further studies to identify and classify forensically relevant insects in Pakistan. There is an exigency demand for further research especially when immature forms of arthropods are recovered from the crime scene.

Keywords: molecular diversity, DNA barcoding, species identification, forensically relevant

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501 Impure Water, a Future Disaster: A Case Study of Lahore Ground Water Quality with GIS Techniques

Authors: Rana Waqar Aslam, Urooj Saeed, Hammad Mehmood, Hameed Ullah, Imtiaz Younas

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This research has been conducted to assess the water quality in and around Lahore Metropolitan area on the basis of three different land uses, i.e. residential, commercial, and industrial land uses. For this, 29 sample sites have been selected on the basis of simple random sampling technique. Samples were collected at the source (WASA tube wells). The criteria for selecting sample sites are to have a maximum concentration of population in the selected land uses. The results showed that in the residential land use the proportion of nitrate and turbidity is at their highest level in the areas of Allama Iqbal Town and Samanabad Town. Commercial land use of Gulberg and Data Gunj Bakhsh Town have highest level of proportion of chlorides, calcium, TDS, pH, Mg, total hardness, arsenic and alkalinity. Whereas in industrial type of land use in Ravi and Wahga Town have the proportion of arsenic, Mg, nitrate, pH, and turbidity are at their highest level. The high rate of concentration of these parameters in these areas is basically due to the old and fractured pipelines that allow bacterial as well as physiochemical contaminants to contaminate the portable water at the sources. Furthermore, it is seen in most areas that waste water from domestic, industrial, as well as municipal sources may get easy discharge into open spaces and water bodies, like, cannels, rivers, lakes that seeps and become a part of ground water. In addition, huge dumps located in Lahore are becoming the cause of ground water contamination as when the rain falls, the water gets seep into the ground and impures the ground water quality. On the basis of the derived results with the help of Geo-spatial technology ACRGIS 9.3 Interpolation (IDW), it is recommended that water filtration plants must be installed with specific parameter control. A separate team for proper inspection has to be made for water quality check at the source. Old water pipelines must be replaced with the new pipelines, and safe water depth must be ensured at the source end.

Keywords: GIS, remote sensing, pH, nitrate, disaster, IDW

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500 Map UI Design of IoT Application Based on Passenger Evacuation Behaviors in Underground Station

Authors: Meng-Cong Zheng

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When the public space is in an emergency, how to quickly establish spatial cognition and emergency shelter in the closed underground space is the urgent task. This study takes Taipei Station as the research base and aims to apply the use of Internet of things (IoT) application for underground evacuation mobility design. The first experiment identified passengers' evacuation behaviors and spatial cognition in underground spaces by wayfinding tasks and thinking aloud, then defined the design conditions of User Interface (UI) and proposed the UI design.  The second experiment evaluated the UI design based on passengers' evacuation behaviors by wayfinding tasks and think aloud again as same as the first experiment. The first experiment found that the design conditions that the subjects were most concerned about were "map" and hoping to learn the relative position of themselves with other landmarks by the map and watch the overall route. "Position" needs to be accurately labeled to determine the location in underground space. Each step of the escape instructions should be presented clearly in "navigation bar." The "message bar" should be informed of the next or final target exit. In the second experiment with the UI design, we found that the "spatial map" distinguishing between walking and non-walking areas with shades of color is useful. The addition of 2.5D maps of the UI design increased the user's perception of space. Amending the color of the corner diagram in the "escape route" also reduces the confusion between the symbol and other diagrams. The larger volume of toilets and elevators can be a judgment of users' relative location in "Hardware facilities." Fire extinguisher icon should be highlighted. "Fire point tips" of the UI design indicated fire with a graphical fireball can convey precise information to the escaped person. "Fire point tips" of the UI design indicated fire with a graphical fireball can convey precise information to the escaped person. However, "Compass and return to present location" are less used in underground space.

Keywords: evacuation behaviors, IoT application, map UI design, underground station

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499 Connotation Reform and Problem Response of Rural Social Relations under the Influence of the Earthquake: With a Review of Wenchuan Decade

Authors: Yanqun Li, Hong Geng

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The occurrence of Wenchuan earthquake in 2008 has led to severe damage to the rural areas of Chengdu city, such as the rupture of the social network, the stagnation of economic production and the rupture of living space. The post-disaster reconstruction has become a sustainable issue. As an important link to maintain the order of rural social development, social network should be an important content of post-disaster reconstruction. Therefore, this paper takes rural reconstruction communities in earthquake-stricken areas of Chengdu as the research object and adopts sociological research methods such as field survey, observation and interview to try to understand the transformation of rural social relations network under the influence of earthquake and its impact on rural space. It has found that rural societies under the earthquake generally experienced three phases: the break of stable social relations, the transition of temporary non-normal state, and the reorganization of social networks. The connotation of phased rural social relations also changed accordingly: turn to a new division of labor on the social orientation, turn to a capital flow and redistribution in new production mode on the capital orientation, and turn to relative decentralization after concentration on the spatial dimension. Along with such changes, rural areas have emerged some social issues such as the alienation of competition in the new industry division, the low social connection, the significant redistribution of capital, and the lack of public space. Based on a comprehensive review of these issues, this paper proposes the corresponding response mechanism. First of all, a reasonable division of labor should be established within the villages to realize diversified commodity supply. Secondly, the villages should adjust the industrial type to promote the equitable participation of capital allocation groups. Finally, external public spaces should be added to strengthen the field of social interaction within the communities.

Keywords: social relations, social support networks, industrial division, capital allocation, public space

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498 Expert Opinions about Barriers to Physical Activity among Ghanaian Adults with Type 2 Diabetes Mellitus: A Qualitative Descriptive Study

Authors: Mohammed Amin, Debra Kerr, Yacoba Atiase, Andrea Driscoll

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Background: Physical activity (PA) is a major component of diabetes self-care management; although this is often stressed to patients, many adults with Type 2 Diabetes Mellitus (T2DM)lead sedentary lifestyles, and barriers exist for uptake of PA. Aim: To explore opinions of healthcare professionals about barriers to PA for adults with T2DM in Ghana. Methods: This qualitative descriptive study included 13healthcare professionals (3 Physiotherapists, 3 exercise physiologists, 3 nurses, and 4 physicians) who provide care to individuals with T2DM in Ghana. Data was collected by semi-structured interviews. The social cognitive theory guided the design of the interview schedule. Data was analysed using thematic analysis. Results: Four main themeswere identified: 1) Individual-related factors, 2) Interpersonal factors, 3) Environment-related factors, and 4) Health system-related factors. Fear of injury, existing co-morbidities, and lack of time make it difficult for people with T2DM to engage in PA. Lack of family support, fear of social ridicule, and cultural beliefs prevent uptake of PA. Poorly designed community spaces, including safe walkways and lack of exercise facilities, inhibit PA participation. Few physical therapists and physical therapy centres exist to support PA participation among people with T2DM. Some nurses and doctors lack adequate knowledge to deliver proper PA education to clients, thereby making clients lack the needed support. Conclusion: Adults with T2DM may be restricted from partaking in PA arising from personal and external factors. This study has identified that barriers to exercise need consideration when designing PA programs that aim to improve health outcomes for people with T2DM in Ghana.

Keywords: type 2 diabetes, physical activity, exercise, ghana, qualitative, barriers

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497 Identification of New Familial Breast Cancer Susceptibility Genes: Are We There Yet?

Authors: Ian Campbell, Gillian Mitchell, Paul James, Na Li, Ella Thompson

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The genetic cause of the majority of multiple-case breast cancer families remains unresolved. Next generation sequencing has emerged as an efficient strategy for identifying predisposing mutations in individuals with inherited cancer. We are conducting whole exome sequence analysis of germ line DNA from multiple affected relatives from breast cancer families, with the aim of identifying rare protein truncating and non-synonymous variants that are likely to include novel cancer predisposing mutations. Data from more than 200 exomes show that on average each individual carries 30-50 protein truncating mutations and 300-400 rare non-synonymous variants. Heterogeneity among our exome data strongly suggest that numerous moderate penetrance genes remain to be discovered, with each gene individually accounting for only a small fraction of families (~0.5%). This scenario marks validation of candidate breast cancer predisposing genes in large case-control studies as the rate-limiting step in resolving the missing heritability of breast cancer. The aim of this study is to screen genes that are recurrently mutated among our exome data in a larger cohort of cases and controls to assess the prevalence of inactivating mutations that may be associated with breast cancer risk. We are using the Agilent HaloPlex Target Enrichment System to screen the coding regions of 168 genes in 1,000 BRCA1/2 mutation-negative familial breast cancer cases and 1,000 cancer-naive controls. To date, our interim analysis has identified 21 genes which carry an excess of truncating mutations in multiple breast cancer families versus controls. Established breast cancer susceptibility gene PALB2 is the most frequently mutated gene (13/998 cases versus 0/1009 controls), but other interesting candidates include NPSR1, GSN, POLD2, and TOX3. These and other genes are being validated in a second cohort of 1,000 cases and controls. Our experience demonstrates that beyond PALB2, the prevalence of mutations in the remaining breast cancer predisposition genes is likely to be very low making definitive validation exceptionally challenging.

Keywords: predisposition, familial, exome sequencing, breast cancer

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496 Unequal Contributions of Parental Isolates in Somatic Recombination of the Stripe Rust Fungus

Authors: Xianming Chen, Yu Lei, Meinan Wang

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The dikaryotic basidiomycete fungus, Puccinia striiformis, causes stripe rust, one of the most important diseases of wheat and barley worldwide. The pathogen is largely reproduced asexually, and asexual recombination has been hypothesized to be one of the mechanisms for the pathogen variations. To test the hypothesis and understand the genetic process of asexual recombination, somatic recombinant isolates were obtained under controlled conditions by inoculating susceptible host plants with a mixture of equal quantity of urediniospores of isolates with different virulence patterns and selecting through a series of inoculation on host plants with different genes for resistance to one of the parental isolates. The potential recombinant isolates were phenotypically characterized by virulence testing on the set of 18 wheat lines used to differentiate races of the wheat stripe rust pathogen, P. striiformis f. sp. tritici (Pst), for the combinations of Pst isolates; or on both sets of the wheat differentials and 12 barley differentials for identifying races of the barley stripe rust pathogen, P. striiformis f. sp. hordei (Psh) for combinations of a Pst isolate and a Psh isolate. The progeny and parental isolates were also genotypically characterized with 51 simple sequence repeat and 90 single-nucleotide polymorphism markers. From nine combinations of parental isolates, 68 potential recombinant isolates were obtained, of which 33 (48.5%) had similar virulence patterns to one of the parental isolates, and 35 (51.5%) had virulence patterns distinct from either of the parental isolates. Of the 35 isolates of distinct virulence patterns, 11 were identified as races that had been previously detected from natural collections and 24 were identified as new races. The molecular marker data confirmed 66 of the 68 isolates as recombinants. The percentages of parental marker alleles ranged from 0.9% to 98.9% and were significantly different from equal proportions in the recombinant isolates. Except for a couple of combinations, the greater or less contribution was not specific to any particular parental isolates as the same parental isolates contributed more to some of the progeny isolates but less to the other progeny isolates in the same combination. The unequal contributions by parental isolates appear to be a general role in somatic recombination for the stripe rust fungus, which may be used to distinguish asexual recombination from sexual recombination in studying the evolutionary mechanisms of the highly variable fungal pathogen.

Keywords: molecular markers, Puccinia striiformis, somatic recombination, stripe rust

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495 In Vivo Investigation of microRNA Expression and Function at the Mammalian Synapse by AGO-APP

Authors: Surbhi Surbhi, Andrea Erni, Gunter Meister, Harold Cremer, Christophe Beclin

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MicroRNAs (miRNAs) are short 20-23 nucleotide long non-coding RNAs; there are 2605 miRNA in humans and 1936 miRNA in mouse in total (miRBase). The nervous system expresses the most abundant miRNA and most diverse. MiRNAs play a role in many steps during neurogenesis, like cell proliferation, differentiation, neural patterning, axon pathfinding, etc. Moreover, in vitro studies suggested a role in the regulation of local translation at the synapse, thus controlling neuronal plasticity. However, due to the specific structure of miRNA molecules, an in-vivo confirmation of the general role of miRNAs in the control of neuronal plasticity is still pending. For example, their small size and their high level of sequence homology make difficult the analysis of their cellular and sub-cellular localization in-vivo by in-situ hybridization. Moreover, it was found that only 40% of the expressed miRNA molecules in a cell are included in RNA-Induced Silencing Complexes (RISC) and, therefore, involved in inhibitory interactions while the rest is silent. Definitively, the development of new tools is needed to have a better understanding of the cellular function of miRNAs, in particular their role in neuronal plasticity. Here we describe a new technique called in-vivo AGO-APP designed to investigate miRNA expression and function in-vivo. This technique is based on the expression of a small peptide derived from the human RISC-complex protein TNRC6B, called T6B, which binds all known Argonaute (Ago) proteins with high affinity allowing the efficient immunoprecipitation of AGO-bound miRNAs. We have generated two transgenic mouse lines conditionally expressing T6B either ubiquitously in the cell or targeted at the synapse. A comparison of the repertoire of miRNAs immuno-precipitated from mature neurons of both mouse lines will provide us with a list of miRNAs showing a specific activity at the synapse. The physiological role of these miRNAs will be subsequently addressed through gain and loss of function experiments.

Keywords: RNA-induced silencing complexes, TNRC6B, miRNA, argonaute, synapse, neuronal plasticity, neurogenesis

Procedia PDF Downloads 114