Search results for: threat detection

Authors: Alessandro Preziosi, Antonio Prioletti, Luca Castangia

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Deformable part models achieve high precision in pedestrian recognition, but all publicly available implementations are too slow for real-time applications. We implemented a deformable part model algorithm fast enough for real-time use by exploiting information about the camera position and orientation. This implementation is both faster and more precise than alternative DPM implementations. These results are obtained by computing convolutions in the frequency domain and using lookup tables to speed up feature computation. This approach is almost an order of magnitude faster than the reference DPM implementation, with no loss in precision. Knowing the position of the camera with respect to horizon it is also possible prune many hypotheses based on their size and location. The range of acceptable sizes and positions is set by looking at the statistical distribution of bounding boxes in labelled images. With this approach it is not needed to compute the entire feature pyramid: for example higher resolution features are only needed near the horizon. This results in an increase in mean average precision of 5% and an increase in speed by a factor of two. Furthermore, to reduce misdetections involving small pedestrians near the horizon, input images are supersampled near the horizon. Supersampling the image at 1.5 times the original scale, results in an increase in precision of about 4%. The implementation was tested against the public KITTI dataset, obtaining an 8% improvement in mean average precision over the best performing DPM-based method. By allowing for a small loss in precision computational time can be easily brought down to our target of 100ms per image, reaching a solution that is faster and still more precise than all publicly available DPM implementations.

Keywords: autonomous vehicles, deformable part model, dpm, pedestrian detection, real time

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Authors: Emir Sunguroglu, Merve Sunguroglu, Yesim Aliefendioglu, Harun Tanrivermis

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Basic human needs range from physiological needs such as food, water and shelter to safety needs such as security, protection from natural disasters and even urban terrorism which are extant and not fulfilled even in urban areas where people live civilly in large communities. These basic needs when arose in urban life lead to a different kind of crime set defined as urban crimes. Urban crimes mostly result from differences between socioeconomic conditions in society. Income inequality increases tendency towards urban crimes. Especially in slum areas and suburbs, urban crimes not only threaten public security but they also affect deliverance of public services. It is highlighted that, construction of urban security against problems caused by urban crimes is not only achieved by involvement of urban security in security of the community but also comprises juridical development and staying above a level of legal standards concurrently. The idea of urban transformation emerged as interventions to demolishment and rebuilding of built environment to solve the unhealthy urban environment, inadequate infrastructure and socioeconomic problems came up during the industrialization process. Considering the probability of urbanization process driving citizens to commit crimes, The United Nations Commission on Human Security’s focus on this theme is conferred to be a proper approach. In this study, the analysis and change in security before, through and after urban transformation, which is one of the tools related to urbanization process, is strived to be discussed through the case of Sincan County Saraycik District. The study also aims to suggest improvements to current legislation on public safety, urban resilience, and urban transformation. In spite of Saraycik District residing in a developing County in Ankara, Turkey, from urbanization perspective as well as socioeconomic and demographic indicators the District exhibits a negative view throughout the County and the country. When related to the county, rates of intentional harm reports, burglary reports, the offense of libel and threat reports and narcotic crime reports are higher. The District is defined as ‘crime hotspot’. Interviews with residents of Saraycik claim that the greatest issue of the neighborhood is Public Order and Security (82.44 %). The District becomes prominent with negative aspects, especially with the presence of unlicensed constructions, occurrence of important social issues such as crime and insecurity and complicated lives of inhabitants from poverty and low standard conditions of living. Additionally, the social structure and demographic properties and crime and insecurity of the field have been addressed in this study. Consequently, it is claimed that urban crime rates were related to level of education, employment and household income, poverty trap, physical condition of housing and structuration, accessibility of public services, security, migration, safety in terms of disasters and emphasized that urban transformation is one of the most important tools in order to provide urban security.

Keywords: urban security, urban crimes, urban transformation, Saraycik district

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Authors: Prittesh Patel, Rushabh Shah, Krishnamurthy Ramar, Vakulbhushan Bhaskar

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The present research work aims at finding genetic differences in the genomes of sugarcane red rot isolates Colletotrichum falcatum Went using Random Amplified Polymorphic DNA (RAPD) and interspersed simple sequence repeat (ISSR) molecular markers. Ten isolates of C. falcatum isolated from different red rot infected sugarcane cultivars stalk were used in present study. The amplified bands were scored across the lanes obtained in 15 RAPD primes and 21 ISSR primes successfully. The data were analysed using NTSYSpc 2.2 software. The results showed 80.6% and 68.07% polymorphism in RPAD and ISSR analysis respectively. Based on the RAPD analysis, ten genotypes were grouped into two major clusters at a cut-off value of 0.75. Geographically distant C. falcatum isolate cfGAN from south Gujarat had a level of similarity with Coimbatore isolate cf8436 presented on separate clade of bootstrapped dendrograms. First and second cluster consisted of five and three isolates respectively, indicating the close relation among them. The 21 ISSR primers produced 119 distinct and scorable loci in that 38 were monomorphic. The number of scorable loci for each primer varied from 2 (ISSR822) to 8 (ISSR807, ISSR823 and ISSR15) with an average of 5.66 loci per primer. Primer ISSR835 amplified the highest number of bands (57), while only 16 bands were obtained by primers ISSR822. Four primers namely ISSR830, ISSR845, ISSR4 and ISSR15 showed the highest value of percentage of polymorphism (100%). The results indicated that both of the marker systems RAPD and ISSR, individually can be effectively used in determination of genetic relationship among C falcatum accessions collected from different parts of south Gujarat.

Keywords: Colletotrichum falcatum, ISSR, RAPD, Red Rot

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Authors: Sanjeewa Seneviratne, Ian Campbell, Nina Scott, Ross Lawrenson

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Introduction: Indigenous Māori women experience a 60% higher breast cancer mortality rate compared with European women in New Zealand. We explored the impact of difference in the rate of screen detected breast cancer between Māori and European women on more advanced disease at diagnosis and lower survival in Māori women. Methods: All primary in-situ and invasive breast cancers diagnosed in screening age women (as defined by the New Zealand National Breast Cancer Screening Programme) between 1999 and 2012 in the Waikato area were identified from the Waikato Breast Cancer Register and the national screening database. Association between screen versus non-screen detection and cancer stage at diagnosis and survival were compared by ethnicity and socioeconomic deprivation. Results: Māori women had 50% higher odds of being diagnosed with more advance staged cancer compared with NZ European women, a half of which was explained by the lower rate of screen detected cancer in Māori women. Significantly lower breast cancer survival rates were observed for Māori compared with NZ European and most deprived compared with most affluent socioeconomic groups for symptomatically detected breast cancer. No significant survival differences by ethnicity or socioeconomic deprivation were observed for screen detected breast cancer. Conclusions: Low rate of screen detected breast cancer appears to be a major contributor for more advanced stage disease at diagnosis and lower breast cancer survival in Māori compared with NZ European women. Increasing screening participation for Māori has the potential to substantially reduce breast cancer mortality inequity between Māori and NZ European women.

Keywords: breast cancer, screening, ethnicity, inequity

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Authors: Reza Soleimanpour, Ching Tai Ng

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Fibre-composites are widely used in various structures due to their attractive properties such as higher stiffness to mass ratio and better corrosion resistance compared to metallic materials. However, one serious weakness of this composite material is delamination, which is a subsurface separation of laminae. A low level of this barely visible damage can cause a significant reduction in residual compressive strength. In the last decade, the application of guided waves for damage detection has been a topic of significant interest for many researches. Among all guided wave techniques, nonlinear guided wave has shown outstanding sensitivity and capability for detecting different types of damages, e.g. cracks and delaminations. So far, most of researches on applications of nonlinear guided wave have been dedicated to isotropic material, such as aluminium and steel, while only a few works have been done on applications of nonlinear characteristics of guided waves in anisotropic materials. This study investigates the nonlinear interactions of the fundamental antisymmetric lamb wave (A0) with delamination in composite laminates using three-dimensional (3D) explicit finite element (FE) simulations. The nonlinearity considered in this study arises from interactions of two interfaces of sub-laminates at the delamination region, which generates contact acoustic nonlinearity (CAN). The aim of this research is to investigate the phenomena of CAN in composite laminated beams by a series of numerical case studies. In this study interaction of fundamental antisymmetric lamb wave with delamination of different sizes are studied in detail. The results show that the A0 lamb wave interacts with the delaminations generating CAN in the form of higher harmonics, which is a good indicator for determining the existence of delaminations in composite laminates.

Keywords: contact acoustic nonlinearity, delamination, fibre reinforced composite beam, finite element, nonlinear guided waves

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Authors: Numair Ahmed Siddiqui, Abdul Hadi bin Abd Rahman, Chow Weng Sum, Wan Ismail Wan Yousif, Asif Zameer, Joel Ben-Awal

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Advances in data capturing from outcrop studies have made possible the acquisition of high-resolution digital data, offering improved and economical reservoir modelling methods. Terrestrial laser scanning utilizing LiDAR (Light detection and ranging) provides a new method to build outcrop based reservoir models, which provide a crucial piece of information to understand heterogeneities in sandstone facies with high-resolution images and data set. This study presents the detailed application of outcrop based sandstone facies connectivity model by acquiring information gathered from traditional fieldwork and processing detailed digital point-cloud data from LiDAR to develop an intermediate small-scale reservoir sandstone facies model of the Miocene Sandakan Formation, Sabah, East Malaysia. The software RiScan pro (v1.8.0) was used in digital data collection and post-processing with an accuracy of 0.01 m and point acquisition rate of up to 10,000 points per second. We provide an accurate and descriptive workflow to triangulate point-clouds of different sets of sandstone facies with well-marked top and bottom boundaries in conjunction with field sedimentology. This will provide highly accurate qualitative sandstone facies connectivity model which is a challenge to obtain from subsurface datasets (i.e., seismic and well data). Finally, by applying this workflow, we can build an outcrop based static connectivity model, which can be an analogue to subsurface reservoir studies.

Keywords: LiDAR, outcrop, high resolution, sandstone faceis, connectivity model

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Authors: Ethan James

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Over one billion people worldwide suffer from some level of vision loss or blindness as a result of progressive retinal diseases. Many patients, particularly in developing areas, are incorrectly diagnosed or undiagnosed whatsoever due to unconventional diagnostic tools and screening methods. Artificial intelligence (AI) based on deep learning (DL) convolutional neural networks (CNN) have recently gained a high interest in ophthalmology for its computer-imaging diagnosis, disease prognosis, and risk assessment. Optical coherence tomography (OCT) is a popular imaging technique used to capture high-resolution cross-sections of retinas. In ophthalmology, DL has been applied to fundus photographs, optical coherence tomography, and visual fields, achieving robust classification performance in the detection of various retinal diseases including macular degeneration, diabetic retinopathy, and retinitis pigmentosa. However, there is no complete diagnostic model to analyze these retinal images that provide a diagnostic accuracy above 90%. Thus, the purpose of this project was to develop an AI model that utilizes machine learning techniques to automatically diagnose specific retinal diseases from OCT scans. The algorithm consists of neural network architecture that was trained from a dataset of over 20,000 real-world OCT images to train the robust model to utilize residual neural networks with cyclic pooling. This DL model can ultimately aid ophthalmologists in diagnosing patients with these retinal diseases more quickly and more accurately, therefore facilitating earlier treatment, which results in improved post-treatment outcomes.

Keywords: artificial intelligence, deep learning, imaging, medical devices, ophthalmic devices, ophthalmology, retina

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Authors: S. Sabatino, V. Calderaro, V. Galdi, G. Graber, L. Ippolito

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Climate change is a rapidly growing global threat caused mainly by increased emissions of carbon dioxide (CO₂) into the atmosphere. These emissions come from multiple sources, including industry, power generation, and the transport sector. The need to tackle climate change and reduce CO₂ emissions is indisputable. A crucial solution to achieving decarbonization in the transport sector is the adoption of electric vehicles (EVs). These vehicles use lithium (Li-Ion) batteries as an energy source, making them extremely efficient and with low direct emissions. However, Li-Ion batteries are not without problems, including the risk of overheating and performance degradation. To ensure its safety and longevity, it is essential to use a battery management system (BMS). The BMS constantly monitors battery status, adjusts temperature and cell balance, ensuring optimal performance and preventing dangerous situations. From the monitoring carried out, it is also able to optimally manage the battery to increase its life. Among the parameters monitored by the BMS, the main ones are State of Charge (SoC), State of Health (SoH), and State of Power (SoP). The evaluation of these parameters can be carried out in two ways: offline, using benchtop batteries tested in the laboratory, or online, using batteries installed in moving vehicles. Online estimation is the preferred approach, as it relies on capturing real-time data from batteries while operating in real-life situations, such as in everyday EV use. Actual battery usage conditions are highly variable. Moving vehicles are exposed to a wide range of factors, including temperature variations, different driving styles, and complex charge/discharge cycles. This variability is difficult to replicate in a controlled laboratory environment and can greatly affect performance and battery life. Online estimation captures this variety of conditions, providing a more accurate assessment of battery behavior in real-world situations. In this article, a hybrid approach based on a neural network and a statistical method for real-time estimation of SoC, SoH, and SoP parameters of interest is proposed. These parameters are estimated from the analysis of a one-day driving profile of an electric vehicle, assumed to be divided into the following four phases: (i) Partial discharge (SoC 100% - SoC 50%), (ii) Partial discharge (SoC 50% - SoC 80%), (iii) Deep Discharge (SoC 80% - SoC 30%) (iv) Full charge (SoC 30% - SoC 100%). The neural network predicts the values of ohmic resistance and incremental capacity, while the statistical method is used to estimate the parameters of interest. This reduces the complexity of the model and improves its prediction accuracy. The effectiveness of the proposed model is evaluated by analyzing its performance in terms of square mean error (RMSE) and percentage error (MAPE) and comparing it with the reference method found in the literature.

Keywords: electric vehicle, Li-Ion battery, BMS, state-of-charge, state-of-health, state-of-power, artificial neural networks

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Authors: Jason M. Hegenauer, Nicholas Fucci

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In the late 1990s, a major deinstitutionalization movement of elderly patients took place, since which, the design of long-term care facilities has not been adequately analyzed in the United States. Over the course of the last 25 years, major innovations in construction methods, technology, and medicine have been developed, drastically changing the landscape of healthcare architecture. In light of recent events, and the expected increase in elderly populations with the aging of the baby-boomer generation, it is evident that reconsideration of these facilities is essential for the proper care of aging populations. The global response has been effective in stifling this pandemic; however, widespread disease still poses an imminent threat to the human race. Having witnessed the devastation Covid-19 has reaped throughout nursing homes and long-term care facilities, it is evident that the current strategies for protecting our most vulnerable populations are not enough. Light renovation of existing facilities and previously overlooked considerations for new construction projects can drastically lower the risk at nursing homes and long-term care facilities. A reconfigured entry sequence supplements several of the features which have been long-standing essentials of the design of these facilities. This research focuses on several aspects identified as needing improvement, including indoor environment quality, security measures incorporated into healthcare architecture and design, and architectural mitigation strategies for sick building syndrome. The results of this study have been compiled as 'best practices' for the design of future healthcare construction projects focused on the health, safety, and quality of life of the residents of these facilities. These design strategies, which can easily be implemented through renovation of existing facilities and new construction projects, minimize risk of infection and spread of disease while allowing routine functions to continue with minimal impact, should the need for future lockdowns arise. Through the current lockdown procedures, which were implemented during the Covid-19 pandemic, isolation of residents has caused great unrest and worry for family members and friends as they are cut off from their loved ones. At this time, data is still being reported, leaving infection and death rates inconclusive; however, recent projections in some states list long-term care facility deaths as high as 60% of all deaths in the state. The population of these facilities consists of residents who are elderly, immunocompromised, and have underlying chronic medical conditions. According to the Centers for Disease Control, these populations are particularly susceptible to infection and serious illness. The obligation to protect our most vulnerable population cannot be overlooked, and the harsh measures recently taken as a response to the Covid-19 pandemic prove that the design strategies currently utilized for doing so are inadequate.

Keywords: building security, healthcare architecture and design, indoor environment quality, new construction, sick building syndrome, renovation

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Authors: K. Mohammed, M. Agarwal, J. Mewman, Y. Ren

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An optimum technic has been developed for extracting volatile organic compounds which contribute to the aroma of lime fruit (Citrus aurantifolia). The volatile organic compounds of healthy and infested lime fruit with California red scale Aonidiella aurantii were characterized using headspace solid phase microextraction (HS-SPME) combined with gas chromatography (GC) coupled flame ionization detection (FID) and gas chromatography with mass spectrometry (GC-MS) as a very simple, efficient and nondestructive extraction method. A three-phase 50/30 μm PDV/DVB/CAR fibre was used for the extraction process. The optimal sealing and fibre exposure time for volatiles reaching equilibrium from whole lime fruit in the headspace of the chamber was 16 and 4 hours respectively. 5 min was selected as desorption time of the three-phase fibre. Herbivorous activity induces indirect plant defenses, as the emission of herbivorous-induced plant volatiles (HIPVs), which could be used by natural enemies for host location. GC-MS analysis showed qualitative differences among volatiles emitted by infested and healthy lime fruit. The GC-MS analysis allowed the initial identification of 18 compounds, with similarities higher than 85%, in accordance with the NIST mass spectral library. One of these were increased by A. aurantii infestation, D-limonene, and three were decreased, Undecane, α-Farnesene and 7-epi-α-selinene. From an applied point of view, the application of the above-mentioned VOCs may help boost the efficiency of biocontrol programs and natural enemies’ production techniques.

Keywords: lime fruit, Citrus aurantifolia, California red scale, Aonidiella aurantii, VOCs, HS-SPME/GC-FID-MS

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Authors: F. Z. Tighilt, N. Belhaneche-Bensemra, S. Belhousse, S. Sam, K. Lasmi, N. Gabouze

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Recently, the gold nanoparticles (Au NPs) became an active multidisciplinary research topic. First, Au thin films fabricated by alkylthiol-functionalized Au NPs were found to have vapor sensitive conductivities, they were hence widely investigated as electrical chemiresistors for sensing different vapor analytes and even organic molecules in aqueous solutions. Second, Au thin films were demonstrated to have speciallocalized surface plasmon resonances (LSPR), so that highly ordered 2D Au superlattices showed strong collective LSPR bands due to the near-field coupling of adjacent nanoparticles and were employed to detect biomolecular binding. Particularly when alkylthiol ligands were replaced by thiol-terminated polymers, the resulting polymer-modified Au NPs could be readily assembled into 2D nanostructures on solid substrates. Monolayers of polystyrene-coated Au NPs showed typical dipolar near-field interparticle plasmon coupling of LSPR. Such polymer-modified Au nanoparticle films have an advantage that the polymer thickness can be feasibly controlled by changing the polymer molecular weight. In this article, the effect of tin-doped indium oxide (ITO) coatings on the plasmonic properties of ITO interfaces modified with gold nanostructures (Au NSs) is investigated. The interest in developing ITO overlayers is multiple. The presence of a con-ducting ITO overlayer creates a LSPR-active interface, which can serve simultaneously as a working electrode in an electro-chemical setup. The surface of ITO/ Au NPs contains hydroxyl groups that can be used to link functional groups to the interface. Here the covalent linking of nickel /Au NSs/ITO hybrid LSPR platforms will be presented.

Keywords: conducting polymer, metal nanoparticles (NPs), LSPR, poly (3-(pyrrolyl)–carboxylic acid), polypyrrole

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Authors: Samia Elfekih, Wee Tek Tay, Karl Gordon, Paul De Barro

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Invasive species represent an increasing threat to food biosecurity, causing significant economic losses in agricultural systems. An example is the sweet potato whitefly, Bemisia tabaci, which is a complex of morphologically indistinguishable species causing average annual global damage estimated at US$2.4 billion. The Bemisia complex represents an interesting model for evolutionary studies because of their extensive distribution and potential for invasiveness and population expansion. Within this complex, two species, Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) have invaded well beyond their home ranges whereas others, such as Indian Ocean (IO) and Australia (AUS), have not. In order to understand why some Bemisia species have become invasive, genome-wide sequence scans were used to estimate population dynamics over time and relate these to climate. The Bayesian Skyline Plot (BSP) method as implemented in BEAST was used to infer the historical effective population size. In order to overcome sampling bias, the populations were combined based on geographical origin. The datasets used for this particular analysis are genome-wide SNPs (single nucleotide polymorphisms) called separately in each of the following groups: Sub-Saharan Africa (Burkina Faso), Europe (Spain, France, Greece and Croatia), USA (Arizona), Mediterranean-Middle East (Israel, Italy), Middle East-Central Asia (Turkmenistan, Iran) and Reunion Island. The non-invasive ‘AUS’ species endemic to Australia was used as an outgroup. The main findings of this study show that the BSP for the Sub-Saharan African MED population is different from that observed in MED populations from the Mediterranean Basin, suggesting evolution under a different set of environmental conditions. For MED, the effective size of the African (Burkina Faso) population showed a rapid expansion ≈250,000-310,000 years ago (YA), preceded by a period of slower growth. The European MED populations (i.e., Spain, France, Croatia, and Greece) showed a single burst of expansion at ≈160,000-200,000 YA. The MEAM1 populations from Israel and Italy and the ones from Iran and Turkmenistan are similar as they both show the earlier expansion at ≈250,000-300,000 YA. The single IO population lacked the latter expansion but had the earlier one. This pattern is shared with the Sub-Saharan African (Burkina Faso) MED, suggesting IO also faced a similar history of environmental change, which seems plausible given their relatively close geographical distributions. In conclusion, populations within the invasive species MED and MEAM1 exhibited signatures of population expansion lacking in non-invasive species (IO and AUS) during the Pleistocene, a geological epoch marked by repeated climatic oscillations with cycles of glacial and interglacial periods. These expansions strongly suggested the potential of some Bemisia species’ genomes to affect their adaptability and invasiveness.

Keywords: whitefly, RADseq, invasive species, SNP, climate change

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Authors: Gagandeep Kaur

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The pesticide industry is a big supplier of agricultural inputs. The uses of pesticides control weeds, fungal diseases, etc., which causes of yield losses in agricultural production. In agribusiness and agrichemical industry, Globalization of markets, competition and innovation are the dominant trends. By the tradition of increasing the productivity of agro-systems through generic, universally applicable technologies, innovation in the agrichemical industry is limited. The marketing of technology of agriculture needs to deal with some various trends such as locally-organized forces that envision regionalized sustainable agriculture in the future. Agricultural production has changed dramatically over the past century. Before World War second agricultural production was featured as a low input of money, high labor, mixed farming and low yields. Although mineral fertilizers were applied already in the second half of the 19th century, most f the crops were restricted by local climatic, geological and ecological conditions. After World War second, in the period of reconstruction, political and socioeconomic pressure changed the nature of agricultural production. For a growing population, food security at low prices and securing farmer income at acceptable levels became political priorities. Current agricultural policy the new European common agricultural policy is aimed to reduce overproduction, liberalization of world trade and the protection of landscape and natural habitats. Farmers have to increase the quality of their productivity and they have to control costs because of increased competition from the world market. Pesticides should be more effective at lower application doses, less toxic and not pose a threat to groundwater. There is a big debate taking place about how and whether to mitigate the intensive use of pesticides. This debate is about the future of agriculture which is sustainable agriculture. This is possible by moving away from conventional agriculture. Conventional agriculture is featured as high inputs and high yields. The use of pesticides in conventional agriculture implies crop production in a wide range. To move away from conventional agriculture is possible through the gradual adoption of less disturbing and polluting agricultural practices at the level of the cropping system. For a healthy environment for crop production in the future there is a need for the maintenance of chemical, physical or biological properties. There is also required to minimize the emission of volatile compounds in the atmosphere. Companies are limiting themselves to a particular interpretation of sustainable development, characterized by technological optimism and production-maximizing. So the main objective of the paper will present the trends in the pesticide industry and in agricultural production in the era of Globalization. The second objective is to analyze sustainable agriculture. Companies of pesticides seem to have identified biotechnology as a promising alternative and supplement to the conventional business of selling pesticides. The agricultural sector is in the process of transforming its conventional mode of operation. Some experts give suggestions to farmers to move towards precision farming and some suggest engaging in organic farming. The methodology of the paper will be historical and analytical. Both primary and secondary sources will be used.

Keywords: globalization, pesticides, sustainable development, organic farming

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Authors: Gachara Grace, Kebira Anthony, Harvey Jagger, Wainaina James

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Aflatoxin contamination of maize in Africa poses a major threat to food security and the health of many African people. In Kenya, aflatoxin contamination of maize is high due to the environmental, agricultural and socio-economic factors. Many studies have been conducted to understand the scope of the problem, especially at pre-harvest level. This research was carried out to gather scientific information on the fungi population, diversity and aflatoxin level during the post-harvest period. The study was conducted in three geographical locations of; Kitui, Kitale and Nakuru. Samples were collected from storage structures of farmers and transported to the Biosciences eastern and central Africa (BecA), International Livestock and Research Institute (ILRI) hub laboratories. Mycoflora was recovered using the direct plating method. A total of five fungal genera (Aspergillus, Penicillium, Fusarium, Rhizopus and Bssyochlamys spp.) were isolated from the stored maize samples. The most common fungal species that were isolated from the three study sites included A. flavus at 82.03% followed by A.niger and F.solani at 49% and 26% respectively. The aflatoxin producing fungi A. flavus was recovered in 82.03% of the samples. Aflatoxin levels were analysed on both the maize samples and in vitro. Most of the A. flavus isolates recorded a high level of aflatoxin when they were analysed for presence of aflatoxin B1 using ELISA. In Kitui, all the samples (100%) had aflatoxin levels above 10ppb with a total aflatoxin mean of 219.2ppb. In Kitale, only 3 samples (n=39) had their aflatoxin levels less than 10ppb while in Nakuru, the total aflatoxin mean level of this region was 239.7ppb. When individual samples were analysed using Vicam fluorometer method, aflatoxin analysis revealed that most of the samples (58.4%) had been contaminated. The means were significantly different (p=0.00<0.05) in all the three locations. Genetic relationships of A. flavus isolates were determined using 13 Simple Sequence Repeats (SSRs) markers. The results were used to generate a phylogenetic tree using DARwin5 software program. A total of 5 distinct clusters were revealed among the genotypes. The isolates appeared to cluster separately according to the geographical locations. Principal Coordinates Analysis (PCoA) of the genetic distances among the 91 A. flavus isolates explained over 50.3% of the total variation when two coordinates were used to cluster the isolates. Analysis of Molecular Variance (AMOVA) showed a high variation of 87% within populations and 13% among populations. This research has shown that A. flavus is the main fungal species infecting maize grains in Kenya. The influence of aflatoxins on human populations in Kenya demonstrates a clear need for tools to manage contamination of locally produced maize. Food basket surveys for aflatoxin contamination should be conducted on a regular basis. This would assist in obtaining reliable data on aflatoxin incidence in different food crops. This would go a long way in defining control strategies for this menace.

Keywords: aflatoxin, Aspergillus flavus, genotyping, Kenya

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Authors: Irfan Ahmad Afip

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This main research objective is to accurately identify the possibility for a Leptospirosis outbreak severity of a certain area based on its input features into a multivariate regression model. The research question is the possibility of an outbreak in a specific area being influenced by this feature, such as social demographics and hydrometeorological extremes. If the occurrence of an outbreak is being subjected to these features, then the epidemic severity for an area will be different depending on its environmental setting because the features will influence the possibility and severity of an outbreak. Specifically, this research objective was three-fold, namely: (a) to identify the relevant multivariate features and visualize the patterns data, (b) to develop a multivariate regression model based from the selected features and determine the possibility for Leptospirosis outbreak in an area, and (c) to compare the predictive ability of multivariate regression model and machine learning algorithms. Several secondary data features were collected locations in the state of Negeri Sembilan, Malaysia, based on the possibility it would be relevant to determine the outbreak severity in the area. The relevant features then will become an input in a multivariate regression model; a linear regression model is a simple and quick solution for creating prognostic capabilities. A multivariate regression model has proven more precise prognostic capabilities than univariate models. The expected outcome from this research is to establish a correlation between the features of social demographic and hydrometeorological with Leptospirosis bacteria; it will also become a contributor for understanding the underlying relationship between the pathogen and the ecosystem. The relationship established can be beneficial for the health department or urban planner to inspect and prepare for future outcomes in event detection and system health monitoring.

Keywords: geographical information system, hydrometeorological, leptospirosis, multivariate regression

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Authors: Ahmed Gaber Abdel Raheem, Nashwa Ahmed Mohamed

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Introduction: Sensorineural hearing loss (SNHL) is largely caused by the degeneration of the cochlea. Therapeutic options for SNHL are limited to hearing aids and cochlear implants. The cell transplantation approach to the regeneration of hair cells has gained considerable attention because stem cells are believed to accumulate in the damaged sites and have the potential for the repair of damaged tissues. The aim of the work: was to assess the use of bone marrow transplantation in repair of damaged inner ear hair cells in rats after the damage had been inflicted by Amikacin injection. Material and Methods: Thirty albino rats were used in this study. They were divided into three groups. Each group ten rats. Group I: used as control. Group II: Were given Amikacin- intratympanic injection till complete loss of hearing function. This could be assessed by Distortion product Otoacoustic Emission (DPOAEs) and / or auditory brain stem evoked potential (ABR). GroupIII: were given intra-peritoneal injection of bone marrow stem cell after complete loss of hearing caused by Amikacin. Clinical assessment was done using DPOAEs and / or auditory brain stem evoked potential (ABR), before and after bone marrow injection. Histological assessment of the inner ear was done by light and electron microscope. Also, Detection of stem cells in the inner ear by immunohistochemistry. Results: Histological examination of the specimens showed promising improvement in the structure of cochlea that may be responsible for the improvement of hearing function in rats detected by DPOAEs and / or ABR. Conclusion: Bone marrow stem cells transplantation might be useful for the treatment of SNHL.

Keywords: amikacin, hair cells, sensorineural hearing loss, stem cells

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Authors: Kanokon Chaicom, Chitima Sirijerachai, Kanchana Chansung, Pinsuda Klangsang, Boonpeng Palaeng, Prajuab Chaimanee, Pimjai Ananta

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Chronic myeloid leukemia (CML) is characterized by the consistent involvement of the Philadelphia chromosome (Ph), which is derived from a reciprocal translocation between chromosome 9 and 22, the main product of the t(9;22) (q34;q11) translocation, is found in the leukemic clone of at least 95% of CML patients. There are two major forms of the BCR/ABL fusion gene, involving ABL exon 2, but including different exons of BCR gene. The transcripts b2a2 (e13a2) or b3a2 (e14a2) code for a p210 protein. Another fusion gene leads to the expression of an e1a2 transcript, which codes for a p190 protein. Other less common fusion genes are b3a3 or b2a3, which codes for a p203 protein and e19a2 (c3a2) transcript, which codes for a p230 protein. Its frequency varies in different populations. In this study, we aimed to report the frequency of BCR-ABL fusion transcript types with CML by multiplex PCR (polymerase chain reaction) in Srinagarind Hospital, Khon Kaen, Thailand. Multiplex PCR for BCR-ABL was performed on 58 patients, to detect different types of BCR-ABL transcripts of the t (9; 22). All patients examined were positive for some type of BCR/ABL rearrangement. The majority of the patients (93.10%) expressed one of the p210 BCR-ABL transcripts, b3a2 and b2a2 transcripts were detected in 53.45% and 39.65% respectively. The expression of an e1a2 transcript showed 3.75%. Co-expression of p210/p230 was detected in 3.45%. Co-expression of p210/p190 was not detected. Multiplex PCR is useful, saves time and reliable in the detection of BCR-ABL transcript types. The frequency of one or other rearrangement in CML varies in different population.

Keywords: chronic myeloid leukemia, BCR-ABL fusion transcript types, multiplex PCR, frequency of BCR-ABL fusion

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Authors: Neda Zamani

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The present study aimed to investigate the effectiveness of training courses and their limitations using the CIPP model. The investigations were done on Isfahan Refinery as a case study. From a purpose point of view, the present paper is included among applied research and from a data gathering point of view, it is included among descriptive research of the field type survey. The population of the study included participants in training courses, their supervisors and experts of the training department. Probability-proportional-to-size (PPS) was used as the sampling method. The sample size for participants in training courses included 195 individuals, 30 supervisors and 11 individuals from the training experts’ group. To collect data, a questionnaire designed by the researcher and a semi-structured interview was used. The content validity of the data was confirmed by training management experts and the reliability was calculated through 0.92 Cronbach’s alpha. To analyze the data in descriptive statistics aspect (tables, frequency, frequency percentage and mean) were applied, and inferential statistics (Mann Whitney and Wilcoxon tests, Kruskal-Wallis test to determine the significance of the opinion of the groups) have been applied. Results of the study indicated that all groups, i.e., participants, supervisors and training experts, absolutely believe in the importance of training courses; however, participants in training courses regard content, teacher, atmosphere and facilities, training process, managing process and product as to be in a relatively appropriate level. The supervisors also regard output to be at a relatively appropriate level, but training experts regard content, teacher and managing processes as to be in an appropriate and higher than average level.

Keywords: training courses, limitations of training effectiveness, CIPP model, Isfahan oil refinery company

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Authors: Levente Kovács, Fruzsina Luca Kézér, Ottó Szenci

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Prediction of the onset of calving and recognizing difficulties at calving has great importance in decreasing neonatal losses and reducing the risk of health problems in the early postpartum period. In this study, changes of rumination time, reticuloruminal pH and temperature were investigated in eutocic (EUT, n = 10) and dystocic (DYS, n = 8) dairy cows around parturition. Rumination time was continuously recorded using an acoustic biotelemetry system, whereas reticuloruminal pH and temperature were recorded using an indwelling and wireless data transmitting system. The recording period lasted from 3 d before calving until 7 days in milk. For the comparison of rumination time and reticuloruminal characteristics between groups, time to return to baseline (the time interval required to return to baseline from the delivery of the calf) and area under the curve (AUC, both for prepartum and postpartum periods) were calculated for each parameter. Rumination time decreased from baseline 28 h before calving both for EUT and DYS cows (P = 0.023 and P = 0.017, respectively). After 20 h before calving, it decreased onwards to reach 32.4 ± 2.3 and 13.2 ± 2.0 min/4 h between 8 and 4 h before delivery in EUT and DYS cows, respectively, and then it decreased below 10 and 5 min during the last 4 h before calving (P = 0.003 and P = 0.008, respectively). Until 12 h after delivery rumination time reached 42.6 ± 2.7 and 51.0 ± 3.1 min/4 h in DYS and EUT dams, respectively, however, AUC and time to return to baseline suggested lower rumination activity in DYS cows than in EUT dams for the 168-h postpartum observational period (P = 0.012 and P = 0.002, respectively). Reticuloruminal pH decreased from baseline 56 h before calving both for EUT and DYS cows (P = 0.012 and P = 0.016, respectively), but did not differ between groups before delivery. In DYS cows, reticuloruminal temperature decreased from baseline 32 h before calving by 0.23 ± 0.02 °C (P = 0.012), whereas in EUT cows such a decrease was found only 20 h before delivery (0.48 ± 0.05 °C, P < 0.01). AUC of reticuloruminal temperature calculated for the prepartum period was greater in EUT cows than in DYS cows (P = 0.042). During the first 4 h after calving, it decreased from 39.7 ± 0.1 to 39.00 ± 0.1 °C and from 39.8 ± 0.1 to 38.8 ± 0.1 °C in EUT and DYS cows, respectively (P < 0.01 for both groups) and reached baseline levels after 35.4 ± 3.4 and 37.8 ± 4.2 h after calving in EUT and DYS cows, respectively. Based on our results, continuous monitoring of changes in rumination time and reticuloruminal temperature seems to be promising in the early detection of cows with a higher risk of dystocia. Depressed postpartum rumination time of DYS cows highlights the importance of the monitoring of cows experiencing difficulties at calving.

Keywords: reticuloruminal pH, reticuloruminal temperature, rumination time, dairy cows, dystocia

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Authors: W. Swiderski

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Composite materials are one answer to the growing demand for materials with better parameters of construction and exploitation. Composite materials also permit conscious shaping of desirable properties to increase the extent of reach in the case of metals, ceramics or polymers. In recent years, composite materials have been used widely in aerospace, energy, transportation, medicine, etc. Fiber-reinforced composites including carbon fiber, glass fiber and aramid fiber have become a major structural material. The typical defect during manufacture and operation is delamination damage of layered composites. When delamination damage of the composites spreads, it may lead to a composite fracture. One of the many methods used in non-destructive testing of composites is active infrared thermography. In active thermography, it is necessary to deliver energy to the examined sample in order to obtain significant temperature differences indicating the presence of subsurface anomalies. To detect possible defects in composite materials, different methods of thermal stimulation can be applied to the tested material, these include heating lamps, lasers, eddy currents, microwaves or ultrasounds. The use of a suitable source of thermal stimulation on the test material can have a decisive influence on the detection or failure to detect defects. Samples of multilayer structure carbon composites were prepared with deliberately introduced defects for comparative purposes. Very thin defects of different sizes and shapes made of Teflon or copper having a thickness of 0.1 mm were screened. Non-destructive testing was carried out using the following sources of thermal stimulation, heating lamp, flash lamp, ultrasound and eddy currents. The results are reported in the paper.

Keywords: Non-destructive testing, IR thermography, composite material, thermal stimulation

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Authors: Matei Tudor Berceanu, Hema Viswambharan, Kirti Kain, Chew Weng Cheng

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Background - Obesity parameters, particularly visceral obesity as measured by the waist-to-height ratio (WHtR), correlate with insulin resistance. The metabolic microvascular changes associated with insulin resistance causes increased peripheral arteriolar resistance primarily to the lower limb vessels. We hypothesize that ankle systolic blood pressures (SBPs) are more significantly associated with visceral obesity than brachial SBPs. Methods - 1098 adults enriched in south Asians or Europeans with diabetes (T2DM) were recruited from a primary care practice in West Yorkshire. Their medical histories, including T2DM and cardiovascular disease (CVD) status, were gathered from an electronic database. The brachial, dorsalis pedis, and posterior tibial SBPs were measured using a Doppler machine. Their body mass index (BMI) and WHtR were calculated after measuring their weight, height, and waist circumference. Linear regressions were performed between the 6 SBPs and both obesity parameters, after adjusting for covariates. Results - Generally, the left posterior tibial SBP (P=4.559*10⁻¹⁵) and right posterior tibial SBP (P=1.114* 10⁻¹³ ) are the pressures most significantly associated with the BMI, as well as in south Asians (P < 0.001) and Europeans (P < 0.001) specifically. In South Asians, although the left (P=0.032) and right brachial SBP (P=0.045) were associated to the WHtR, the left posterior tibial SBP (P=0.023) showed the strongest association. Conclusion - Regardless of ethnicity, ankle SBPs are more significantly associated with generalized obesity than brachial SBPs, suggesting their screening potential for screening for early detection of T2DM and CVD. A combination of ankle SBPs with WHtR is proposed in south Asians.

Keywords: ankle blood pressures, body mass index, insulin resistance, waist-to-height-ratio

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Authors: N. G. Vepkhvadze, T. Enukidze

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Anthrax is a fatal disease caused by strains of Bacillus anthracis, a spore-forming gram-positive bacillus that causes the disease anthrax in animals and humans. Anthrax is a zoonotic disease that is also well-recognized as a potential agent of bioterrorism. Infection in humans is extremely rare in the developed world and is generally due to contact with infected animals or contaminated animal products. Testing of this zoonotic disease began in 1907 in Georgia and is still being tested routinely to provide accurate information and efficient testing results at the State Laboratory of Agriculture of Georgia. Each clinical sample is analyzed by RT-PCR and bacteriology methods; this study used Real-Time PCR assays for the detection of B. anthracis that rely on plasmid-encoded targets with a chromosomal marker to correctly differentiate pathogenic strains from non-anthracis Bacillus species. During the period of 2015-2022, the State Laboratory of Agriculture (SLA) tested 250 clinical and environmental (soil) samples from several different regions in Georgia. In total, 61 out of the 250 samples were positive during this period. Based on the results, Anthrax cases are mostly present in Eastern Georgia, with a high density of the population of livestock, specifically in the regions of Kakheti and Kvemo Kartli. All laboratory activities are being performed in accordance with International Quality standards, adhering to biosafety and biosecurity rules by qualified and experienced personnel handling pathogenic agents. Laboratory testing plays the largest role in diagnosing animals with anthrax, which helps pertinent institutions to quickly confirm a diagnosis of anthrax and evaluate the epidemiological situation that generates important data for further responses.

Keywords: animal disease, baccilus anthracis, edp, laboratory molecular diagnostics

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Authors: Mirele Plenishti

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Revisionist powers are sometimes associated to revolutionary and status quo powers, this because along the line representing the level of satisfaction–dissatisfaction with the system, revisionist powers are located in between status quo and revolutionary powers. In particular, the case of revisionist powers seeking for social status adjustments (while having status quo intentions) can, in the first option, be refuted due to the disbelief that dissatisfaction could coexist with status quo intentions – this entailing the possibility to trigger a spiral effect by over-counter-reacting. In the second option, revisionist powers can be underestimated as a real threat, this entailing a potential inadequate reaction. The necessity to well manage international change entails the need to understand better how revisionist powers seek for changes in status, within the system. The complexity of this case is heightened by the propensity of both IR scholars and practitioners to infer states' aims and intentions – towards the system – by looking at their behaviours. This has resulted in the tendency to consider cooperative international behaviours as symptomatic of status quo intentions, and vice versa: status quo intentions as manifested through positive/cooperative behaviours. Similarly, assertive/competitive international behaviours are considered as symptomatic (and vice versa, as manifestations) of revolutionary intentions. Therefore, within complex and composite foreign policies, scholars who disbelieve the existence of revisionist powers with status quo intentions, tend to highlight the negative/competitive elements; while more optimist scholars tend to focus on conforming/cooperative behaviours. Both perspectives, while understanding relevant components of the complex international interaction, still miss a composite overview. In order to closely investigate the strategies adopted by (status quo aiming) revisionist states, and by drawing on sociological studies on peer relations, focused on children's behaviour, one could expect that the compresence of both positive (compliant/cooperative) and negative (competitive/assertive) behaviours, is deliberate, and functional to seeking social status adjustments. Indeed, at the end of 90s, peer relation studies focused on children's behaviour, discerned between the concept of social acceptance (that refers to the degree of social preference assigned to the child– how much is s/he liked) and popularity (which refers to the social status assigned to the child within the group). By building on this distinction, it was possible to identify a link relating social acceptance to prosocial (compliant/cooperative) behaviours and strategies, and popularity to both prosocial and antisocial (aggressive/assertive) behaviours and strategies. Since then, antisocial behaviours ceased to be considered as a proof of social maladjustment and were finally identified as socially recognized strategies adopted in function of the achievement of popularity. Drawing on these results, one can hypothesize that also international status seekers perform both positive (conforming/compliant/cooperative) and negative (assertive/aggressive/competitive) behaviours. Therefore, the link between aims and behaviours loses its strength, since cooperative and competitive behaviours are both means for status seeking strategies that aim at status quo intentions. By carrying out a historical investigation of Italy's foreign policy during fascism, the intent is to closely look at this compresence of behaviours, in order to better qualify its components and their relations.

Keywords: compresence of cooperative and competitive behaviours and strategies, revisionist powers, status quo intentions, status seeking

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Authors: Clemence Collon, Didier Poracchia

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The manipulation of sports competitions is a new type of sports integrity problem. While doping has become an organized, institutionalized struggle, the manipulation of sports competitions is gradually building up. This study aims to describe and understand how the soft Olympic and Paralympic law was gradually built. It also summarizes the legal tools for prevention, detection, and sanction developed by the international Olympic movement. Then, it analyzes the impact of this soft law on the law of the States, in particular in French law. This study is mainly based on an analysis of existing legal literature and non-binding law in the International Olympic and Paralympic movement and on the French National Olympic Committee. Interviews were carried out with experts from the Olympic movement or experts working on combating the manipulation of sports competitions; the answers are also used in this article. The International Olympic Committee has created a supranational legal base to fight against the manipulation of sports competitions. This legal basis must be respected by sports organizations. The Olympic Charter, the Olympic Code of Ethics, the Olympic Movement Code on the prevention of the manipulation of sports competitions, the rules of standards, the basic universal principles, the manuals, the declarations have been published in this perspective. This sports soft law has influences or repercussions in each state. Many states take this new form of integrity problem into account by creating state laws or measures in favor of the fight against sports manipulations. France has so far only a legal basis for manipulation related to betting on sports competitions through the infraction of sports corruption included in the penal code and also created a national platform with various actors to combat this cheating. This legal study highlights the progressive construction of the sports law rules of the Olympic movement in the fight against the manipulation of sports competitions linked to sports betting and their impact on the law of the states.

Keywords: integrity, law and ethics, manipulation of sports competitions, olympic, sports law

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Authors: Janos Juhasz, Sandor Pongor, Balazs Ligeti

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Next-generation sequencing, especially whole genome shotgun sequencing, is becoming a common approach to gain insight into the microbiomes in a culture-independent way, even in clinical practice. It does not only give us information about the species composition of an environmental sample but opens the possibility to detect antimicrobial resistance and novel, or currently unknown, pathogens. Accurately and reliably detecting the microbial strains is a challenging task. Here we present a sensitive approach for detecting pathogens in metagenomics samples with special regard to detecting novel variants of known pathogens. We have developed a pipeline that uses fast, short read aligner programs (i.e., Bowtie2/BWA) and comprehensive nucleotide databases. Taxonomic binning is based on the lowest common ancestor (LCA) principle; each read is assigned to a taxon, covering the most significantly hit taxa. This approach helps in balancing between sensitivity and running time. The program was tested both on experimental and synthetic data. The results implicate that our method performs as good as the state-of-the-art BLAST-based ones, furthermore, in some cases, it even proves to be better, while running two orders magnitude faster. It is sensitive and capable of identifying taxa being present only in small abundance. Moreover, it needs two orders of magnitude less reads to complete the identification than MetaPhLan2 does. We analyzed an experimental anthrax dataset (B. anthracis strain BA104). The majority of the reads (96.50%) was classified as Bacillus anthracis, a small portion, 1.2%, was classified as other species from the Bacillus genus. We demonstrate that the evaluation of high-throughput sequencing data is feasible in a reasonable time with good classification accuracy.

Keywords: metagenomics, taxonomy binning, pathogens, microbiome, B. anthracis

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Authors: Bahareh Golchin, Nooshin Riahi

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One of the most significant issues as attended a lot in recent years is that of recognizing the sentiments and emotions in social media texts. The analysis of sentiments and emotions is intended to recognize the conceptual information such as the opinions, feelings, attitudes and emotions of people towards the products, services, organizations, people, topics, events and features in the written text. These indicate the greatness of the problem space. In the real world, businesses and organizations are always looking for tools to gather ideas, emotions, and directions of people about their products, services, or events related to their own. This article uses the Twitter social network, one of the most popular social networks with about 420 million active users, to extract data. Using this social network, users can share their information and opinions about personal issues, policies, products, events, etc. It can be used with appropriate classification of emotional states due to the availability of its data. In this study, supervised learning and deep neural network algorithms are used to classify the emotional states of Twitter users. The use of deep learning methods to increase the learning capacity of the model is an advantage due to the large amount of available data. Tweets collected on various topics are classified into four classes using a combination of two Bidirectional Long Short Term Memory network and a Convolutional network. The results obtained from this study with an average accuracy of 93%, show good results extracted from the proposed framework and improved accuracy compared to previous work.

Keywords: emotion classification, sentiment analysis, social networks, deep neural networks

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Authors: Cheng Zeng, George Michailidis, Hitoshi Iyatomi, Leo L. Duan

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The conditional density characterizes the distribution of a response variable y given other predictor x and plays a key role in many statistical tasks, including classification and outlier detection. Although there has been abundant work on the problem of Conditional Density Estimation (CDE) for a low-dimensional response in the presence of a high-dimensional predictor, little work has been done for a high-dimensional response such as images. The promising performance of normalizing flow (NF) neural networks in unconditional density estimation acts as a motivating starting point. In this work, the authors extend NF neural networks when external x is present. Specifically, they use the NF to parameterize a one-to-one transform between a high-dimensional y and a latent z that comprises two components [zₚ, zₙ]. The zₚ component is a low-dimensional subvector obtained from the posterior distribution of an elementary predictive model for x, such as logistic/linear regression. The zₙ component is a high-dimensional independent Gaussian vector, which explains the variations in y not or less related to x. Unlike existing CDE methods, the proposed approach coined Augmented Posterior CDE (AP-CDE) only requires a simple modification of the common normalizing flow framework while significantly improving the interpretation of the latent component since zₚ represents a supervised dimension reduction. In image analytics applications, AP-CDE shows good separation of 𝑥-related variations due to factors such as lighting condition and subject id from the other random variations. Further, the experiments show that an unconditional NF neural network based on an unsupervised model of z, such as a Gaussian mixture, fails to generate interpretable results.

Keywords: conditional density estimation, image generation, normalizing flow, supervised dimension reduction

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Authors: Peng Hou

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Natural products produced from marine bacteria serve as an immense reservoir for anti-infective drugs and therapeutic agents. Nowadays, heterologous expression of gene clusters of interests has been widely adopted as an effective strategy for natural product discovery. Briefly, the heterologous expression flowchart would be: biosynthetic gene cluster identification, pathway construction and expression, and product detection. However, gene cluster capture using traditional Transformation-associated recombination (TAR) protocol is low-efficient (0.5% positive colony rate). To make things worse, most of these putative new natural products are only predicted by bioinformatics analysis such as antiSMASH, and their corresponding natural products biosynthetic pathways are either not expressed or expressed at very low levels under laboratory conditions. Those setbacks have inspired us to focus on seeking new technologies to efficiently edit and refractor of biosynthetic gene clusters. Recently, two cutting-edge techniques have attracted our attention - the CRISPR-Cas9 and Gibson Assembly. By now, we have tried to pretreat Brevibacillus laterosporus strain genomic DNA with CRISPR-Cas9 nucleases that specifically generated breaks near the gene cluster of interest. This trial resulted in an increase in the efficiency of gene cluster capture (9%). Moreover, using Gibson Assembly by adding/deleting certain operon and tailoring enzymes regardless of end compatibility, the silent construct (~80kb) has been successfully refactored into an active one, yielded a series of analogs expected. With the appearances of the novel molecular tools, we are confident to believe that development of a high throughput mature pipeline for DNA assembly, transformation, product isolation and identification would no longer be a daydream for marine natural product discovery.

Keywords: biosynthesis, CRISPR-Cas9, DNA assembly, refactor, TAR cloning

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Authors: Lina T. Al Kury, Oleg I. Voitychuk, Ramiz M. Ali, Sehamuddin Galadari, Keun-Hang Susan Yang, Frank Christopher Howarth, Yaroslav M. Shuba, Murat Oz

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A role for anandamide (N-arachidonoyl ethanolamide; AEA), a major endocannabinoid, in the cardiovascular system in various pathological conditions has been reported in earlier studies. In the present work, we have hypothesized that the antiarrhythmic effects reported for AEA are due to its negative inotropic effect and altered action potential (AP) characteristics. Therefore, we tested the effects of AEA on contractility and electrophysiological properties of rat ventricular myocytes. Video edge detection was used to measure myocyte shortening. Intracellular Ca2+ was measured in cells loaded with the fluorescent indicator fura-2 AM. Whole-cell patch-clamp technique was employed to investigate the effect of AEA on the characteristics of APs. AEA (1 μM) caused a significant decrease in the amplitudes of electrically-evoked myocyte shortening and Ca2+ transients and significantly decreased the duration of AP. The effect of AEA on myocyte shortening and AP characteristics was not altered in the presence of pertussis toxin (PTX, 2 µg/ml for 4 h), AM251 and SR141716 (cannabinoid type 1 receptor antagonists) or AM630 and SR 144528 (cannabinoid type 2 receptor antagonists). Furthermore, AEA inhibited voltage-activated inward Na+ (INa) and Ca2+ (IL,Ca) currents; major ionic currents shaping the APs in ventricular myocytes, in a voltage and PTX-independent manner. Collectively, the results suggest that AEA depresses ventricular myocyte contractility, by decreasing the action potential duration (APD), and inhibits the function of voltage-dependent Na+ and L-type Ca2+ channels in a manner independent of cannabinoid receptors. This mechanism may be importantly involved in the antiarrhythmic effects of anandamide.

Keywords: action potential, anandamide, cannabinoid receptor, endocannabinoid, ventricular myocytes

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Authors: Tiantian Min, Chuanxiang Cheng, Jin Yue

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The growth and reproduction of microorganisms in food packaging can cause food decay and foodborne diseases, which pose a serious threat to the health of consumers and even cause serious economic losses. Active food packaging containing antibacterial bioactive compounds is a promising strategy for extending the shelf life of products and maintaining the food quality, as well as reducing the food waste. However, most active packaging can only act as slow-release effect for antimicrobials, which causes the release rate of antimicrobials not match the growth rate of microorganisms. Stimuli-responsive active packaging materials based on biopolymeric substrates and bioactive substances that respond to some biological and non-biological trigger factors provide more opportunities for fresh food preservation. The biological stimuli factors such as relative humidity, pH and enzyme existed in the exudate secreted by microorganisms have been expected to design food packaging materials. These stimuli-responsive materials achieved accurate release or delivery of bioactive substances at specific time and appropriate dose. Recently, metal-organic-frameworks (MOFs) nanoparticles become attractive carriers to enhance the efficiency of bioactive compounds or drugs. Cellulose nanofibrils have been widely applied for film substrates due to their biodegradability and biocompatibility. The abundant hydroxyl groups in cellulose can be oxidized to carboxyl groups by TEMPO, making it easier to anchoring MOFs and to be further modification. In this study, a pH and enzyme dual-responsive CAR@ZIF-8/TOCNF/PE film was fabricated by in-situ growth of ZIF-8 nanoparticles onto TEMPO-oxidized cellulose (TOCNF) film and further coated with pectin (PE) for stabilization and controlled release of carvacrol (CAR). The enzyme triggered release of CAR was achieved owing to the degradation of pectin by pectinase secreted by microorganisms. Similarly, the pH-responsive release of CAR was attributed to the unique skeleton degradation of ZIF-8, further accelerating the release of CAR from the topological structure of ZIF-8. The composite film performed excellent crystallinity and adsorb ability confirmed by X-ray diffraction and BET analysis, and the inhibition efficiency against Escherichia coli, Staphylococcus aureus and Aspergillus niger reached more than 99%. The composite film was capable of releasing CAR when exposure to dose-dependent enzyme (0.1, 0.2, and 0.3 mg/mL) and acidic condition (pH = 5). When inoculated 10 μL of Aspergillus niger spore suspension on the equatorial position of mango and raspberries, this composite film acted as packaging pads effectively inhibited the mycelial growth and prolonged the shelf life of mango and raspberries to 7 days. Such MOF-TOCNF based film provided a targeted, controlled and sustained release of bioactive compounds for long-term antibacterial activity and preservation effect, which can also avoid the cross-contamination of fruits.

Keywords: active food packaging, controlled release, fruit preservation, in-situ growth, stimuli-responsive

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