Search results for: reference gene

Authors: Vitalijs Lusis, Andrejs Krasnikovs

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Fiber reinforced concrete is important material for load bearing structural elements. Usually fibers are homogeneously distributed in a concrete body having arbitrary spatial orientations. At the same time, in many situations, fiber concrete with oriented fibers is more optimal. Is obvious, that is possible to create constructions with oriented short fibers in them, in different ways. Present research is devoted to one of such approaches- fiber reinforced concrete prisms having dimensions 100 mm×100 mm×400 mm with layers of non-homogeneously distributed fibers inside them were fabricated. Simultaneously prisms with homogeneously dispersed fibers were produced for reference as well. Prisms were tested under four point bending conditions. During the tests vertical deflection at the center of every prism and crack opening were measured (using linear displacements transducers in real timescale). Prediction results were discussed.

Keywords: fiber reinforced concrete, 4-point bending, steel fiber, construction engineering

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Authors: Artem Kim, Clara Savary, Christele Dubourg, Wilfrid Carre, Houda Hamdi-Roze, Valerie Dupé, Sylvie Odent, Marie De Tayrac, Veronique David

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Holoprosencephaly (HPE) is a rare congenital brain malformation resulting from the incomplete separation of the two cerebral hemispheres. It is characterized by a wide phenotypic spectrum and a high degree of locus heterogeneity. Genetic defects in 16 genes have already been implicated in HPE, but account for only 30% of cases, suggesting that a large part of genetic factors remains to be discovered. HPE has been recently redefined as a complex multigenic disorder, requiring the joint effect of multiple mutational events in genes belonging to one or several developmental pathways. The onset of HPE may result from accumulation of the effects of multiple rare variants in functionally-related genes, each conferring a moderate increase in the risk of HPE onset. In order to decipher the genetic basis of HPE, unconventional patterns of inheritance involving multiple genetic factors need to be considered. The primary objective of this study was to uncover possible disease causing combinations of multiple rare variants underlying HPE by performing trio-based Whole Exome Sequencing (WES) of familial cases where no molecular diagnosis could be established. 39 families were selected with no fully-penetrant causal mutation in known HPE gene, no chromosomic aberrations/copy number variants and without any implication of environmental factors. As the main challenge was to identify disease-related variants among a large number of nonpathogenic polymorphisms detected by WES classical scheme, a novel variant prioritization approach was established. It combined WES filtering with complementary gene-level approaches: transcriptome-driven (RNA-Seq data) and clinically-driven (public clinical data) strategies. Briefly, a filtering approach was performed to select variants compatible with disease segregation, population frequency and pathogenicity prediction to identify an exhaustive list of rare deleterious variants. The exome search space was then reduced by restricting the analysis to candidate genes identified by either transcriptome-driven strategy (genes sharing highly similar expression patterns with known HPE genes during cerebral development) or clinically-driven strategy (genes associated to phenotypes of interest overlapping with HPE). Deeper analyses of candidate variants were then performed on a family-by-family basis. These included the exploration of clinical information, expression studies, variant characteristics, recurrence of mutated genes and available biological knowledge. A novel bioinformatics pipeline was designed. Applied to the 39 families, this final integrated workflow identified an average of 11 candidate variants per family. Most of candidate variants were inherited from asymptomatic parents suggesting a multigenic inheritance pattern requiring the association of multiple mutational events. The manual analysis highlighted 5 new strong HPE candidate genes showing recurrences in distinct families. Functional validations of these genes are foreseen.

Keywords: complex genetic disorder, holoprosencephaly, multiple rare variants, whole exome sequencing

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Authors: Osama Terra, Hatem Hussein, Adriaan Van Brakel

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Dense wavelength division multiplexing (DWDM) technology requires tight specification and therefore measurement of wavelength accuracy and stability of the telecommunication lasers. Thus, calibration of the used Optical Spectrum Analyzers (OSAs) that are used to measure wavelength is of a great importance. Proficiency testing must be performed on such measuring activity to insure the accuracy of the measurement results. In this paper, a new comparison scheme is introduced to test the performance of such calibrations. This comparison scheme is implemented between NIS-Egypt and NMISA-South Africa for the calibration of the wavelength scale of an OSA. Both institutes employ reference gas cell to calibrate OSA according to the standard IEC/ BS EN 62129 (2006). The result of this comparison is compiled in this paper.

Keywords: OSA calibration, HCN gas cell, DWDM technology, wavelength measurement

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Authors: Apotiola Z. O., Anyakorah C. I., Kuforiji O. O.

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Sensory and microbial properties of fresh and canned Calocybe indica (milky mushroom) were evaluated. The mushroom was grown under a controlled environment with hardwood (Cola nitida) and rice bran substrate (4:1) canned in a brine solution of salt and citric acid. Analysis was carried out using standard methods. The overall acceptability ranged between 5.62 and 6.50, with sample S30 adjudged the best. In all, significant differences p<0.01 exist in the panelist judgment. Thus, the incorporation of salt and citric acid at 3.5g and 1.5g, respectively, improved sensory attributes such as texture, aroma, color, and overall acceptability. There was no coliform and fungi growth on the samples throughout the storage period. The bacterial count, on the other hand, was observed only in the fifth and sixth week of the storage period which varied between 0.2 to 0.9 x 103 cfu/g. The highest value was observed in sample S20 of the sixth week of storage, while the lowest value was recorded in sample S30 of the sixth week of storage. Based on 16S rRNA gene sequencing, bacterial species were taxonomically confirmed as Bacillus thuringiensis. The percentile compositions and Sequence ID of the bacterial species in the mushroom was 90%.

Keywords: bacterial count, microbial property, sensory, sawdust, texture

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Authors: Maryam Ghasemi

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One of the most significant components that provide comfort and protection is having a shelter called a house. Even if components and regions are changed or restored to meet new functions, the house's identity must be preserved. In the contemporary era, houses are increasingly being built regardless of cultural identity. This misunderstanding caused a sense of unease. This study analyses archaic and modern architecture to find semiotic areas and qualities in the latter, using the former as a reference. This study's technique used an exploratory assessment of architectural components from both periods. The Abbasid residence and the Ekbatan architectural complex were used as case studies. The identity of Iranian architecture does not correlate with current buildings. The other part is privacy, which is a missing link between traditional and modern Iranian architecture because it is directly related to the identities of homes based on the cultures of their residents.

Keywords: housing, traditional, contemporary, privacy, semiotic

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Authors: Xinyu Gong, Zhilin Li, Xintao Liu

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Maps are a means of communication. Cartographic language involves established theories of natural language for understanding maps. “Cartographic words’, or “map symbols”, are crucial elements of cartographic language. Personalized mapping is increasingly popular, with growing demands for customized map-making by the general public. Automated symbol-making and customization play a key role in personalized mapping. However, formal representations for the automated construction of map symbols are still lacking. In natural language, the process of word and sentence construction can be formalized. Through the analogy between natural language and graphical language, formal representations of natural language construction can be used as a reference for constructing cartographic language. We selected Chinese character structures (i.e., S

Keywords: personalized mapping, Chinese character, cartographic language, map symbols

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Authors: Manal Farea, Adam Husein, Ahmad S. Halim, Zurairah Berahim, Nurul A. Abdullah, Khairani I. Mokhtar, Kasmawati Mokhtar

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Endodontic furcal perforation remains both an endodontic and a periodontal problem. Regeneration of cementum is very essential for the perforation repair. The aim of this study was to investigate the role of Hertwig's epithelial root sheath (HERS) cells on the cementogenic differentiation of stem cells derived from human exfoliated deciduous teeth (SHED) in the presence of chitosan scaffold-TGFβ1. HERS cells were isolated and characterized then co-cultured with SHED with/without chitosan scaffold-TGFβ1. SHED proliferation was assessed by PrestoBlue. Alkaline phosphatase activity, mineralization behaviour and gene/protein expression of cemento/osteoblast phenotype of SHED were evaluated. Results of the present study showed that HERS cells in association with chitosan-TGFβ1 enhanced proliferation and cemento/osteogenic differentiation of SHED. Our novel co-culture system confirmed the potential effect of HERS cells to stimulate the differentiation of SHED along the cementoblastic lineage which was triggered in the presence of chitosan-TGFβ1. This approach possesses a novel therapeutic strategy for future endodontic perforation and periodontitis.

Keywords: cementogenesis, co-culture system, HERS, SHED

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Authors: Gong Kangming, Zhao Caiqi

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High-rise special-shaped structure, such as main frame structure of the statues, is one of the structure forms in irregular structure widely used. Due to the complex shape of the statue structure, with a large aspect ratio, its wind load value and the overall mechanical properties are very different from the high-rise buildings with the general rules. The paper taking a certain 48 meters high main frame structure of the statue located in Shuangyashan City, Heilongjiang Province, static and dynamic properties are analyzed by the finite element software. Through static and dynamic analysis, it got a number of useful conclusions that have a certain reference value for the analysis and design of the future similar structure.

Keywords: a Buddhism goddess Guanyin body, wind load, dynamic analysis, bolster, node design

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Authors: A. Ali, V. Strezov, P. Davies, I. Wright, T. Kan

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The environmental impacts arising from mining activities affect the air, water, and soil quality. Impacts may result in unexpected and adverse environmental outcomes. This study reports on the impact of coal production on sediment in Sydney region of Australia. The sediment samples upstream and downstream from the discharge points from three mines were taken, and 80 parameters were tested. The results were assessed against sediment quality based on presence of metals. The study revealed the increment of metal content in the sediment downstream of the reference locations. In many cases, the sediment was above the Australia and New Zealand Environment Conservation Council and international sediment quality guidelines value (SQGV). The major outliers to the guidelines were nickel (Ni) and zinc (Zn).

Keywords: coal mine, environmental impact, produced water, sediment quality guidelines value (SQGV)

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Authors: Sunil Verma

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In today’s world non-renewable sources are depleting quickly, so there is a need to produce efficient and unconventional engines to minimize the use of fuel. Also, there are many fatal accidents happening every year during extraction, distillation, transportation and storage of fuel. Reason for explosions of gaseous fuel is unwanted autoignition. Autoignition characterstics of fuel are mandatory to study to build efficient engines and to avoid accidents. This report is concerned with study of autoignition delay characteristics of iso-octane by using rapid compression machine. The paper clearly explains the dependence of ignition delay characteristics on variation of equivalence ratios from lean to rich mixtures. The equivalence ratio is varied from 0.3 to 1.2.

Keywords: autoignition, iso-octane, combustion, rapid compression machine, equivalence ratio, ignition delay

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Authors: Maria Grazia Albanesi, Riccardo Amadeo

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This study presents a new model of the human image quality assessment process: the aim is to highlight the foundations of the image quality metrics proposed in literature, by identifying the cognitive/physiological or mathematical principles of their development and the relation with the actual human quality assessment process. The model allows to create a novel categorization of objective and subjective image quality metrics. Our work includes an overview of the most used or effective objective metrics in literature, and, for each of them, we underline its main characteristics, with reference to the rationale of the proposed model and categorization. From the results of this operation, we underline a problem that affects all the presented metrics: the fact that many aspects of human biases are not taken in account at all. We then propose a possible methodology to address this issue.

Keywords: eye-tracking, image quality assessment metric, MOS, quality of user experience, visual perception

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Authors: Safa Ougoujil, Sidi Mohamed Rigar

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Economics is not an exact science. It cannot be from the moment it is a social science that concerns society organization, a human science that depends on the behavior of the men and women who make a part of this society. Therefore, it cannot ignore morality, the instinctive sense of good and evil, the natural order which place us between certain values, and which religion often sheds light on. In terms of finance, the reference to ethics is becoming more popular than ever. This is naturally due to the growing financial crises. Finance is less and less ethical, but some financial practices have continued to do so. This is the case of ethical finance and Islamic finance. After attempting to define the concepts of ethical finance and Islamic finance, in a period when financial innovation seeks to encourage differentiation in order to create more profit margins, this article attempts to expose the particularities, the convergences and the potentialities of development of these two sensibilities.

Keywords: convergences, ethical finance, Islamic finance, potential development

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Authors: Safiullah Abdullahi

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This paper describes the design of a digital control system which controls the speed and direction of a robot. The robot is expected to follow a black thick line with the highest possible speed and lowest error around the line. The control system of the robot will correct for the angle error that is made between the frame axis of the robot and the line. The cause for error is the difference in speed of the two driving wheels of the robot which are driven by two separate DC motors, whereas the speed difference in wheels is due to the un-modeled fraction that is available in the wheels with different magnitudes in each. The control scheme is that a number of photo sensors are mounted in the front of the robot and report their position in reference to the black line to the digital controller. The controller then, evaluates the position error and generates the needed duty cycle for the related wheel motor to drive it faster or slower.

Keywords: digital control, robot, controller, control system

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Authors: Jie Yang, Qifu Cheng, Yafang Liu, Zhijie Gu

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Controlled emissions of effluent from nuclear power plants are an important means of ensuring environmental safety. In order to fully grasp the actual discharge level of nuclear power plant in China's nuclear power plant in the pressurized water reactor and heavy water reactor, it will use the global average nuclear power plant effluent discharge as a reference to the standard analysis of China's nuclear power plant environmental discharge status. The results show that the average normalized emission of liquid tritium in PWR nuclear power plants in China is slightly higher than the global average value, and the other nuclides emissions are lower than the global average values.

Keywords: radioactive effluent, HWR, PWR, nuclear power plant

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Authors: I. Muslih, A. Alkhalailah, A. Merdji

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Photovoltaic efficiency is highly affected by dust accumulation; the dust particles prevent direct solar radiation from reaching the panel surface; therefore a reduction in output power will occur. A study of dust and soiling accumulation effect on the output power of PV panels was conducted for different periods of time from May to October in three countries of the MENA region, Jordan, Egypt, and Algeria, under local weather conditions. This study leads to build a more realistic equation to estimate the power reduction as a function of time. This logarithmic function shows the high reduction in power in the first days with 10% reduction in output power compared to the reference system, where it reaches a steady state value after 60 days to reach a maximum value of 30%.

Keywords: dust effect, MENA, solar energy, PV system

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Authors: Doaa Hashad, Amany Elbanna, Abeer Ibrahim, Gihan Khedr

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Background: H19 is one of the long non coding RNAs (LncRNA) that is related to the progression of many diseases including cancers. This work was carried out to study the level of the long non-coding RNA; H119, in plasma of patients with gastric cancer (GC) and to assess its significance in their clinical management. Methods: A total of sixty-two participants were enrolled in the present study. The first group included thirty-two GC patients, while the second group was formed of thirty age and sex matched healthy volunteers serving as a control group. Plasma samples were used to assess H19 gene expression using real time quantitative PCR technique. Results: H19 expression was up-regulated in GC patients with positive correlation to TNM cancer stages. Conclusions: Up-regulation of H19 is closely associated with gastric cancer and correlates well with tumor staging. Convenient, efficient quantification of H19 in plasma using real time PCR technique implements its role as a potential noninvasive prognostic biomarker in gastric cancer, that predicts patient’s outcome and most importantly as a novel target in gastric cancer treatment with better performance achieved on using both CEA and H19 simultaneously.

Keywords: biomarker, gastric, cancer, LncRNA

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Authors: M. N. Hanina, M. Hairul Shahril, I. Ismatul Nurul Asyikin, A. K. Abdul Jalil, M. R. Salina, M. R. Maryam, M. Rosfarizan

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The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Keywords: Bacillus subtilis ATCC21332, streptomycin sulfate, extracellular proteins, antibiotics

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Authors: Khiem M. Nguyen, Ming C. Yang

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Chloroplast pigments are extremely important during photosynthesis since they play essential roles in light absorption and energy transfer. Therefore, understanding the efficiency of chlorophyll (Chl) biosynthesis could facilitate enhancement in photo-assimilates accumulation, and ultimately, in crop yield. The Chl-deficient mutants have been used extensively to study the Chl biosynthetic pathways and the biogenesis of the photosynthetic apparatus. Rice (Oryza sativa L.) is one of the most leading food crops, serving as staple food for many parts of the world. To author’s best knowledge, Chl b–lacking rice has been found; however the molecular mechanism of Chl biosynthesis still remains unclear compared to wild-type rice. In this study, the ultrastructure analysis, photosynthetic properties, and transcriptome profile of wild-type rice (Norin No.8, N8) and its Chl b-lacking mutant (Chlorina 1, C1) were examined. The finding concluded that total Chl content and Chl b content in the C1 leaves were strongly reduced compared to N8 leaves, suggesting that reduction in the total Chl content contributes to leaf color variation at the physiological level. Plastid ultrastructure of C1 possessed abnormal thylakoid membranes with loss of starch granule, large number of vesicles, and numerous plastoglobuli. The C1 rice also exhibited thinner stacked grana, which was caused by a reduction in the number of thylakoid membranes per granum. Thus, the different Chl a/b ratio of C1 may reflect the abnormal plastid development and function. Transcriptional analysis identified 23 differentially expressed genes (DEGs) and 671 transcription factors (TFs) that were involved in Chl metabolism, chloroplast development, cell division, and photosynthesis. The transcriptome profile and DEGs revealed that the gene encoding PsbR (PSII core protein) was down-regulated, therefore suggesting that the lower in light-harvesting complex proteins are responsible for the lower photosynthetic capacity in C1. In addition, expression level of cell division protein (FtsZ) genes were significantly reduced in C1, causing chloroplast division defect. A total of 19 DEGs were identified based on KEGG pathway assignment involving Chl biosynthesis pathway. Among these DEGs, the GluTR gene was down-regulated, whereas the UROD, CPOX, and MgCH genes were up-regulated. Observation through qPCR suggested that later stages of Chl biosynthesis were enhanced in C1, whereas the early stages were inhibited. Plastid structure analysis together with transcriptomic analysis suggested that the Chl a/b ratio was amplified both by the reduction in Chl contents accumulation, owning to abnormal chloroplast development, and by the enhanced conversion of Chl b to Chl a. Moreover, the results indicated the same Chl-cycle pattern in the wild-type and C1 rice, indicating another Chl b degradation pathway. Furthermore, the results demonstrated that normal grana stacking, along with the absence of Chl b and greatly reduced levels of Chl a in C1, provide evidence to support the conclusion that other factors along with LHCII proteins are involved in grana stacking. The findings of this study provide insight into the molecular mechanisms that underlie different Chl a/b ratios in rice.

Keywords: Chl-deficient mutant, grana stacked, photosynthesis, RNA-Seq, transcriptomic analysis

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Authors: Fernando Duarte

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The work aims to evaluate the use of classical image processing methodologies towards skin tone classification in dermatological images. The skin tone is an important attribute when considering several factor for skin cancer diagnosis. Currently, there is a lack of clear methodologies to classify the skin tone based only on the dermatological image. In this work, a recent released dataset with the label for skin tone was used as reference for the evaluation of classical methodologies for segmentation and adjustment of color space for classification of skin tone in dermatological images. It was noticed that even though the classical methodologies can work fine for segmentation and color adjustment, classifying the skin tone without proper control of the aquisition of the sample images ended being very unreliable.

Keywords: segmentation, classification, color space, skin tone, Fitzpatrick

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Authors: N. A. Kazemi Sefat, M. M. Mohammadi, J. Hadjati, S. Talebi, M. Ajami, H. Daneshvar

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Colorectal cancer metastases result in a significant number of cancer related deaths. Histone deacetylase (HDAC) inhibitors induce growth arrest and apoptosis in a variety of human cancer cells. Sodium butyrate (SB) is a short chain fatty acid, belongs to HDAC inhibitors which is released in the colonic lumen as a consequence of fiber fermentation. In this study, we are about to assess the effect of sodium butyrate on HCT116 human colorectal carcinoma cell line. The viability of cells was measured by microscopic morphologic study and MTT assay. After 48 hours, treatments more than 10 mM lead to cell injury in HCT116 by increasing cell granulation and decreasing cell adhesion (p>0.05). After 72 hours, treatments at 10 mM and more lead to significant cell injury (p<0.05). Our results may suggest that the gene expression which is contributed in cell proliferation and apoptosis has been changed under pressure of HDAC inhibition.

Keywords: colorectal cancer, sodium butyrate, cytotoxicity, MTT

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Authors: Chang Ting, Wu Chun Kuan

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Wireless sensing technology is increasingly developed, in order to avoid caregiver neglect children in poor physiological condition, so there are more and more products into the wireless sensor-related technologies, in order to reduce the risk of infants. In view of this, the study will focus on Somatosensory detection wristbands Applied Research of Baby, and to explore through observation and literature, to find design criteria which conform baby products, as well as the advantages and disadvantages of existing products. This study will focus on 0-2 years of infant research and product design, to provide 2-3 new design concepts and products to identify weaknesses through the use of the actual product, further provide future baby wristbands design reference.

Keywords: infants, observation, design criteria, wireless sensing

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Authors: Jung-Min Yang

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Corrective control enables us to change the stable state behavior of an asynchronous sequential machine without modifying inner logic of the machine. This paper addresses corrective control for asynchronous machines with switching capability. The considered asynchronous machine consists of a set of different submachines and switches to each machine according to a constant switching sequence. The control goal is to design a corrective controller such that the closed-loop system can match the behavior of a reference model. The reachability of the switched asynchronous machine is described by a logic calculation of the reachability of submachines. The design procedure of the proposed corrective controller is outlined, and the applicability of the proposed scheme is validated in an example.

Keywords: switched asynchronous sequential machines, corrective control, state feedback, switching sequences

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Authors: Napat Watjanatepin, Wikorn Wong-Satiean

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The objective of this research is to design and construct the PV-Wind hybrid autonomous system for the greenhouse plantation, and analyze the technical performance of the PV-Wind energy system. This design depends on the water consumption in the greenhouse by using 24 of the fogging mist each with the capability of 24 liter/min. The operating time is 4 times per day, each round for 15 min. The fogging system is being driven by water pump with AC motor rating 0.5 hp. The load energy consumed is around 1.125 kWh/d. The designing results of the PV-Wind hybrid energy system is that sufficient energy could be generated by this system. The results of this study can be applied as a technical data reference for other areas in the central part of Thailand.

Keywords: PV-Wind hybrid autonomous system, greenhouse plantation, fogging system, central part of Thailand

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Authors: Hsiu-Hua Cheng

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In recent years, the number of social-oriented virtual communities users has increased significantly. Corporate investment in advertising on social-oriented virtual communities increases quickly. With the gigantic commercial value of the digital market, competitions between virtual communities are keen. In this context, how to retain existing customers to continue using social-oriented virtual communities is an urgent issue for virtual community managers. This study employs the perspective of automaticity mechanism and combines the social embeddedness theory with the literature of involvement and habit in order to explore antecedents of users’ continuous usage on social-oriented virtual communities. The results can be a reference for scholars and managers of social-oriented virtual communities.

Keywords: continued usage, habit, social embeddedness, involvement, virtual community

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Authors: Jisong Zhang, Yinghua Zhao

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The present study investigated the mechanical properties and microstructure of Ultra-High Performance Concrete (UHPC) containing supplementary cementitious materials (SCMs), such as fly ash (FA) and silica fume (SF), and to verify the synergistic effect in the ternary system. On the basis of 30% fly ash replacement, the incorporation of either 10% SF or 20% SF show a better performance compared to the reference sample. The efficiency factor (k-value) was calculated as a synergistic effect to predict the compressive strength of UHPC with these SCMs. The SEM of micrographs and pore volume from BJH method indicate a high correlation with compressive strength. Further, an artificial neural networks model was constructed for prediction of the compressive strength of UHPC containing these SCMs.

Keywords: artificial neural network, fly ash, mechanical properties, ultra-high performance concrete

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Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

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Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

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Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

Abstract:

Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

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Authors: Mohamed A. Dkhil, Denis Delic, Saleh Al-Quraishy

Abstract:

Coccidiosis causes considerable economic loss in the poultry industry. The current study aimed to investigate the response of goblet cells as well as the induced tissue damage during Eimeria papilata infection. Mice were infected with sporulated E. papillata oocyts. On day 5 post-infection, the fecal output was determined. Also, the jejunum was prepared for the histological, histochemical and molecular studies. Our results revealed that the intestinal coccidian infection with E. papillata induced a marked goblet cell hypoplasia and depleted mucus secretion. Also, the infection was able to alter the jejuna architecture and increased the apoptotic cells inside the villi. In addition, the real time PCR results indicated that, the inflammatory cytokines TNF-α, iNOS, IFN-y and IL-1β were significantly up-regulated. In contrast, the mRNA expression patterns of IL-6 in response to E. papillata infection did not differ significantly between control and infected mice. Moreover, the mRNA expression of TLR4 was significantly up-regulated, whereas the expression of MUC2 is significantly down-regulated upon infection. Further studies are required to understand the regulatory mechanisms of goblet cells related genes.

Keywords: goblet cells, Eimeria papillata, mice, jejunum

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Authors: Jahanshah Ashkani, Kevin J. Naidoo

Abstract:

Glycosylation is the major posttranslational modification (PTM) process in cellular development. In tumour development, it is marked by structural alteration of carbohydrates (glycans) that is the result of aberrant glycosylation. Altered glycan structures affect cell surface ligand-receptor interactions that interfere with the regulation of cell adhesion, migration, and proliferation. The resulting changes in glycan biosynthesis pathways originate from altered expression of glycosyltransferases and glycosidases. While the alteration in glycosylation patterns is a recognized “hallmark of cancer”, the influential overview of the biology of cancer proposes eight hallmarks with no explicit suggestion to connectivity with glycosylation. Recently, we have discovered a connection between the glycosyltransferase gene expression and cancer type and subtype. Here we present an association between aberrant glycosylation and the biological hallmarks of breast cancer by exploring the common regulatory mechanisms at the genomic scale. The result of this study bridges the glycobiological and biological pathways that are accepted hallmarks of cancer by connecting their common regulatory pathways. This is an impetus for further investigation as target therapies of breast cancer are very likely to be uncovered from this.

Keywords: aberrant glycosylation, biological hallmarks, breast cancer, regulatory mechanism

Procedia PDF Downloads 249

Authors: Olga V. Chervyakova, Elmira T. Tailakova, Vitaliy M. Strochkov, Kulyaisan T. Sultankulova, Nurlan T. Sandybayev, Lev G. Nemchinov, Rosemarie W. Hammond

Abstract:

Sheep pox is a highly contagious infection that OIE regards to be one of the most dangerous animal diseases. It causes enormous economic losses because of death and slaughter of infected animals, lower productivity, cost of veterinary and sanitary as well as quarantine measures. To control spread of sheep pox infection the attenuated vaccines are widely used in the Republic of Kazakhstan and other Former Soviet Union countries. In spite of high efficiency of live vaccines, the possible presence of the residual virulence, potential genetic instability restricts their use in disease-free areas that leads to necessity to exploit new approaches in vaccine development involving recombinant DNA technology. Vaccines on the basis of recombinant proteins are the newest generation of prophylactic preparations. The main advantage of these vaccines is their low reactogenicity and this fact makes them widely used in medical and veterinary practice for vaccination of humans and farm animals. The objective of the study is to produce recombinant immunogenic proteins for development of the high-performance means for sheep pox prophylaxis. The SPV proteins were chosen for their homology with the known immunogenic vaccinia virus proteins. Assay of nucleotide and amino acid sequences of the target SPV protein genes. It has been shown that four proteins SPPV060 (ortholog L1), SPPV074 (ortholog H3), SPPV122 (ortholog A33) and SPPV141 (ortholog B5) possess transmembrane domains at N- or C-terminus while in amino acid sequences of SPPV095 (ortholog А 4) and SPPV117 (ortholog А 27) proteins these domains were absent. On the basis of these findings the primers were constructed. Target genes were amplified and subsequently cloned into the expression vector рЕТ26b(+) or рЕТ28b(+). Six constructions (pSPPV060ΔТМ, pSPPV074ΔТМ, pSPPV095, pSPPV117, pSPPV122ΔТМ and pSPPV141ΔТМ) were obtained for expression of the SPV genes under control of T7 promoter in Escherichia coli. To purify and detect recombinant proteins the amino acid sequences were modified by adding six histidine molecules at C-terminus. Induction of gene expression by IPTG was resulted in production of the proteins with molecular weights corresponding to the estimated values for SPPV060, SPPV074, SPPV095, SPPV117, SPPV122 and SPPV141, i.e. 22, 30, 20, 19, 17 and 22 kDa respectively. Optimal protocol of expression for each gene that ensures high yield of the recombinant protein was identified. Assay of cellular lysates by western blotting confirmed expression of the target proteins. Recombinant proteins bind specifically with antibodies to polyhistidine. Moreover all produced proteins are specifically recognized by the serum from experimentally SPV-infected sheep. The recombinant proteins SPPV060, SPPV074, SPPV117, SPPV122 and SPPV141 were also shown to induce formation of antibodies with virus-neutralizing activity. The results of the research will help to develop a new-generation high-performance means for specific sheep pox prophylaxis that is one of key moments in animal health protection. The research was conducted under the International project ISTC # K-1704 “Development of methods to construct recombinant prophylactic means for sheep pox with use of transgenic plants” and under the Grant Project RK MES G.2015/0115RK01983 "Recombinant vaccine for sheep pox prophylaxis".

Keywords: prophylactic preparation, recombinant protein, sheep pox virus, subunit vaccine

Procedia PDF Downloads 237