Search results for: visual expression
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3830

Search results for: visual expression

2420 The Angiogenic Activity of α-Mangostin in the Development of Zebrafish Embryo In Vivo

Authors: Titis Indah Adi Rahayu

Abstract:

Angiogenesis is the process of generating new capillary from pre-existing blood vessels. VEGFA is a major regulator in angiogenesis that binds and activates two tyrosine kinase receptors, VEGFR1 (Flt-1) and VEGFR2 (Flk-1/KDR) which regulate pathological and physiological angiogenesis. Disruption of VEGFA and VEGFR2 regulation lead to many diseases. The study of α-Mangostin (derivate of xanthone) as anti-oxidant and anti inflammation has been explored recently and both of them have relation to vasculature however the effect of α-Mangostin in blood vessel formation in healthy tissue in vivo has not been studied. Zebrafish is a powerful model in studying angiogenesis and shared many advantages that is a viable whole animal model for screening small molecules that affect blood vessel formation. Therefore the aim of this study is to evaluate angiogenic activity of α-Mangostin in healthy tissue in vivo in zebrafish embryo in relation of patterning blood vessel. Blood vessel patterning is highly characteristic in the developing of zebrafish embryo and the subintestinal vessel (SIV) can be stained and visualized microscopically as a primary screen for α-Mangostin that effect angiogenesis. The zebrafish embryos are divided into 2 groups. Group one consists of the zebrafish embryos at 1 dpf for 4 days which are tested to α-Mangostin in several concentration 2 µM, 4 µM, 6 µM, 8 µM and 10 µM whereas in group two the zebrafish larva at 4 dpf are exposed to α-Mangostin 1,75 µM, 2,3 µM, 2,9 µM, 3,8 µM dan 5 µM for 2 days. DMSO is served as a control for each group. The level expression of vegfa and vegfr2 are observed quantitatively using real time q-PCR and patterning of SIV are then analized via alkaline phospatase staining. Result shows that the level expression of vegfa and vegfr2 is repressed quantitatively as shown in real time q-PCR in the group of 1-4 days of α-Mangostin exposure where it is increased in the group of 4-6 days of α-Mangostin exposure. The result is then compared to alkaline phospatase staining of SIV using stereo microscope. It indicates that α-Mangostin does not disturb the patterning of SIV formation in zebrafish.

Keywords: angiogenesis, Danio rerio, α-Mangostin, SIV, vegfa, vegfr2

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2419 Human Dental Pulp Stem Cells Attenuate Streptozotocin-Induced Parotid Gland Injury in Rats

Authors: Gehan ElAkabawy

Abstract:

Background: Diabetes mellitus causes severe deteriorations of almost all the organs and systems of the body, as well as significant damage to the oral cavity. The oral changes are mainly related to salivary glands dysfunction characterized by hyposalivation and xerostomia, which significantly reduce diabetic patients’ quality of life. Human dental pulp stem cells represent a promising source for cell-based therapies, owing to their easy, minimally invasive surgical access, and high proliferative capacity. It was reported that the trophic support mediated by dental pulp stem cells can rescue the functional and structural alterations of damaged salivary glands. However, potential differentiation and paracrine effects of human dental pulp stem cells in diabetic-induced parotid gland damage have not been previously investigated. Our study aimed to investigate the therapeutic effects of intravenous transplantation of human dental pulp stem cells (hDPSCs) on parotid gland injury in a rat model of streptozotocin (STZ)-induced type 1 diabetes. Methods: Thirty Sprague-Dawley male rats were randomly categorised into three groups: control, diabetic (STZ), and transplanted (STZ+hDPSCs). hDPSCs or vehicle was injected into the tail vein 7 days after STZ injection. The fasting blood glucose levels were monitored weekly. A glucose tolerance test was performed, and the parotid gland weight, salivary flow rate, oxidative stress indices, parotid gland histology, and caspase-3, vascular endothelial growth factor (VEGF), and proliferating cell nuclear antigen (PCNA) expression in parotid tissues were assessed 28 days post-transplantation. Results: Transplantation of hDPSCs downregulated blood glucose, improved the salivary flow rate, and reduced oxidative stress. The cells migrated to, survived, and differentiated into acinar, ductal, and myoepithelial cells in the STZ-injured parotid gland. Moreover, they downregulated the expression of caspase-3 and upregulated the expression of VEGF and PCNA, likely exerting pro-angiogenetic and antiapoptotic effects and promoting endogenous regeneration. In addition, the transplanted cells enhanced the parotid nitric oxide (NO) -tetrahydrobiopterin (BH4) pathway. Conclusions: Our results show that hDPSCs can migrate to and survive within the STZ-injured parotid gland, where they prevent its functional and morphological damage by restoring normal glucose levels, differentiating into parotid cell populations, and stimulating paracrine-mediated regeneration. Thus, hDPSCs may have therapeutic potential in the treatment of diabetes-induced parotid gland injury.

Keywords: dental pulp stem cells, diabetes, streptozotocin, parotid gland

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2418 Spatial Indeterminacy: Destabilization of Dichotomies in Modern and Contemporary Architecture

Authors: Adrian Lo

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Since the beginning of modern architecture, ideas of free plan and transparency have proliferated well into current trends of building design, from houses to highrise office buildings. The movement’s notion of a spatially homogeneous, open, and limitless ‘free plan’ stands opposite to the spatially heterogeneous ‘separation of rooms’ defined by load-bearing walls, which in turn triggered new notions of transparency achieved by vast expanses of glazed walls. Similarly, transparency was also dichotomized as something that was physical or optical, as well as something conceptual, akin to spatial organization. As opposed to merely accepting the duality and possible incompatibility of these dichotomies, this paper seeks to ask how can space be both literally and phenomenally transparent, as well as display both homogeneous and heterogeneous qualities? This paper explores this potential destabilization or blurring of spatial phenomena by dissecting the transparent layers and volumes of a series of selected case studies to investigate how different architects have devised strategies of spatial ambivalence, ambiguity, and interpenetration. Projects by Peter Eisenman, Sou Fujimoto, and SANAA will be discussed and analyzed to show how the superimposition of geometries and spaces achieve different conditions of layering, transparency, and interstitiality. Their particular buildings will be explored to reveal various innovative kinds of spatial interpenetration produced through the articulate relations of the elements of architecture, which challenge conventional perceptions of interior and exterior whereby visual homogeneity blurs with spatial heterogeneity. The results show how spatial conceptions such as interpenetration and transparency have the ability to subvert not only inside-outside dialectics but could also produce multiple degrees of interiority within complex and indeterminate spatial dimensions in constant flux as well as present alternative forms of social interaction.

Keywords: interpenetration, literal and phenomenal transparency, spatial heterogeneity, visual homogeneity

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2417 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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2416 A Five–Year Review Study of Epidemiology of Ocular and Adnexal Injuries Requiring Surgical Intervention in a Middle Eastern Area: Al Ain, UAE

Authors: Tahra AlMahmoud, Sameeha Mohamed Al Hadhrami, Mohamed Elhanan, Hanan Naser Alshamsi, Fikri Abu-Zidan

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Background: To the best of the author(s)’ knowledge there are no epidemiological studies for traumatic eye injuries in UAE, neither data on groups at risk or mechanisms for ocular trauma. Purpose: To report the epidemiology of eye injuries that required hospital admission and surgery at a referral center at the eastern part of Abu Dhabi. Method: Retrospective charts review of all patients who had suffered an eye injury that required surgical intervention between 2012 and 2017 at Al Ain Hospital. Demographic data, place of occurrence, the cause of injury, visual acuity (VA) before and after treatment, number of admission days and follow up were extracted. Data were tabulated and presented as number (%), mean (SD), or median (range) as appropriate. Wilcoxon signed rank test was used for VA outcome. Results: One hundred forty-one patients were identified, 96 eyes with open-globe and 48 other types of injuries. The mean age of the patients was 26±15.5 years, and 89% were male. Majority of injuries occurred at the workplace (50.4%) followed by home (31.2%). Trauma with a sharp object (24.1%), blunt object (16.3%), nail (11.3%), and hammer on metal (7.8%) were the most common etiologies of injury. Corneas injuries (48.2%) was the most frequent cause for visual acuity limitation followed by lens/cataract (23.4%). Among the traumatized eyes, 30 eyes (21.3%) retained intraocular foreign body, Mean admission days was 3.16± 2.81days (1-16) and a number of follow up visit was 3.17± 4.11times (0-26). Conclusion: Ocular trauma requiring surgical intervention is an area of concern in particular for occupations involving work with metals. This work may give insight into the value and necessity of implementing preventive measures.

Keywords: epidemiology, Middle Eastern area, occupational injury, ocular traumas

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2415 Proteomics Associated with Colonization of Human Enteric Pathogen on Solanum lycopersicum

Authors: Neha Bhadauria, Indu Gaur, Shilpi Shilpi, Susmita Goswami, Prabir K. Paul

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The aerial surface of plants colonized by Human Enteric Pathogens ()has been implicated in outbreaks of enteric diseases in humans. Practice of organic farming primarily using animal dung as manure and sewage water for irrigation are the most significant source of enteric pathogens on the surface of leaves, fruits and vegetables. The present work aims to have an insight into the molecular mechanism of interaction of Human Enteric Pathogens or their metabolites with cell wall receptors in plants. Tomato plants grown under aseptic conditions at 12 hours L/D photoperiod, 25±1°C and 75% RH were inoculated individually with S. fonticola and K. pneumonia. The leaves from treated plants were sampled after 24 and 48 hours of incubation. The cell wall and cytoplasmic proteins were extracted and isocratically separated on 1D SDS-PAGE. The sampled leaves were also subjected to formaldehyde treatment prior to isolation of cytoplasmic proteins to study protein-protein interactions induced by Human Enteric Pathogens. Protein bands extracted from the gel were subjected to MALDI-TOF-TOF MS analysis. The foremost interaction of Human Enteric Pathogens on the plant surface was found to be cell wall bound receptors which possibly set ups a wave a critical protein-protein interaction in cytoplasm. The study revealed the expression and suppression of specific cytoplasmic and cell wall-bound proteins, some of them being important components of signaling pathways. The results also demonstrated HEP induced rearrangement of signaling pathways which possibly are crucial for adaptation of these pathogens to plant surface. At the end of the study, it can be concluded that controlling the over-expression or suppression of these specific proteins rearrange the signaling pathway thus reduces the outbreaks of food-borne illness.

Keywords: cytoplasmic protein, cell wall-bound protein, Human Enteric Pathogen (HEP), protein-protein interaction

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2414 Protective Effect of Bexarotene, a Selective RXRα Agonist, against Hypotension Associated with Inflammation and Tissue Injury Linked to Decreased Circulating iNOS Levels in A Rat Model of Septic Shock

Authors: Bahar Tunctan, Sefika Pinar Kucukkavruk, Meryem Temiz-Resitoglu, Demet Sinem Guden, Ayse Nihal Sari, Seyhan Sahan-Firat

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We hypothesized that rexinoids such as bexarotene, a selective retinoid X receptor α (RXRα) agonist, may be beneficial for preventing mortality due to inflammation associated with increased expression/activity of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide (LPS). Therefore, we investigated effects of bexarotene on the changes in circulating protein levels of iNOS (an index for systemic iNOS expression), myeloperoxidase (MPO) (an index for systemic inflammation), and lactate dehydrogenase (LDH) (an index for systemic tissue injury) in LPS-induced systemic inflammation model resulting in septic shock in rats. Rats were injected with saline (4 ml/kg; i.p.), LPS (10 mg/kg; i.p.), dimethylsulphoxide (4 ml/kg, 0.1%; s.c.) at time 0. Mean arterial blood pressure and heart rate were measured using a tail-cuff device. Bexarotene (0.03, 0.1, 0.3, and 1 mg/kg; s.c.) was administered to separate groups of rats 1 h after injection of saline or LPS. The rats were sacrificed 4 h after saline or LPS injection and blood was collected for measurement of serum iNOS, MPO, and LDH protein levels. Blood pressure decreased by 31 mmHg and heart rate increased by 63 bpm in the LPS-treated rats. Bexarotene at 0.3 and 1 mg/kg doses caused 20% mortality 4 h after LPS injection. In the LPS-treated rats, serum iNOS, MPO, and LDH protein levels were increased. Bexarotene only at 0.1 mg/kg dose prevented the LPS-induced hypotension and increased in iNOS, MPO, and LDH protein levels. These data are consistent with the view that a decrease in systemic iNOS levels contributes to the beneficial effect of bexarotene to prevent the hypotension associated with inflammation and tissue injury during rat endotoxemia. [This work was financially supported by The Scientific and Technological Research Council of Turkey (SBAG-109S121)].

Keywords: bexarotene, inflammation, iNOS, lipopolisaccharide, RXRa

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2413 Signed Language Phonological Awareness: Building Deaf Children's Vocabulary in Signed and Written Language

Authors: Lynn Mcquarrie, Charlotte Enns

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The goal of this project was to develop a visually-based, signed language phonological awareness training program and to pilot the intervention with signing deaf children (ages 6 -10 years/ grades 1 - 4) who were beginning readers to assess the effects of systematic explicit American Sign Language (ASL) phonological instruction on both ASL vocabulary and English print vocabulary learning. Growing evidence that signing learners utilize visually-based signed language phonological knowledge (homologous to the sound-based phonological level of spoken language processing) when reading underscore the critical need for further research on the innovation of reading instructional practices for visual language learners. Multiple single-case studies using a multiple probe design across content (i.e., sign and print targets incorporating specific ASL phonological parameters – handshapes) was implemented to examine if a functional relationship existed between instruction and acquisition of these skills. The results indicated that for all cases, representing a variety of language abilities, the visually-based phonological teaching approach was exceptionally powerful in helping children to build their sign and print vocabularies. Although intervention/teaching studies have been essential in testing hypotheses about spoken language phonological processes supporting non-deaf children’s reading development, there are no parallel intervention/teaching studies exploring hypotheses about signed language phonological processes in supporting deaf children’s reading development. This study begins to provide the needed evidence to pursue innovative teaching strategies that incorporate the strengths of visual learners.

Keywords: American sign language phonological awareness, dual language strategies, vocabulary learning, word reading

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2412 Investigating Role of Novel Molecular Players in Forebrain Roof-Plate Midline Invagination

Authors: Mohd Ali Abbas Zaidi, Meenu Sachdeva, Jonaki Sen

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In the vertebrate embryo, the forebrain anlagen develops from the anterior-most region of the neural tube which is the precursor of the central nervous system (CNS). The roof plate located at the dorsal midline region of the forebrain anlagen, acts as a source of several secreted molecules involved in patterning and morphogenesis of the forebrain. One such key morphogenetic event is the invagination of the forebrain roof plate which results in separation of the single forebrain vesicle into two cerebral hemispheres. Retinoic acid (RA) signaling plays a key role in this process. Blocking RA signaling at the dorsal forebrain midline inhibits dorsal invagination and results in the absence of certain key features of this region, such as thinning of the neuroepithelium and a lowering of cell proliferation. At present we are investigating the possibility of other signaling pathways acting in concert with RA signaling to regulate this process. We have focused on BMP signaling, which we found to be active in a mutually exclusive domain to that of RA signaling within the roof plate. We have also observed that there is a change in BMP signaling activity on modulation of RA signaling indicating an antagonistic relationship between the two. Moreover, constitutive activation of BMP signaling seems to completely inhibit thinning and partially affect invagination, leaving the lowering of cell proliferation in the midline unaffected. We are employing in-silico modeling as well as molecular manipulations to investigate the relative contribution if any, of regional differences in rates of cell proliferation and thinning of the neuroepithelium towards the process of invagination. We have found expression of certain cell adhesion molecules in forebrain roof-plate whose mRNA localization across the thickness of neuroepithelium is influenced by Bmp and RA signaling, giving regional rigidity to roof plate and assisting invagination. We also found expression of certain cytoskeleton modifiers in a localized small domains in invaginating forebrain roof plate suggesting that midline invagination is under control of many factors.

Keywords: bone morphogenetic signaling, cytoskeleton, cell adhesion molecules, forebrain roof plate, retinoic acid signaling

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2411 Detecting Memory-Related Gene Modules in sc/snRNA-seq Data by Deep-Learning

Authors: Yong Chen

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To understand the detailed molecular mechanisms of memory formation in engram cells is one of the most fundamental questions in neuroscience. Recent single-cell RNA-seq (scRNA-seq) and single-nucleus RNA-seq (snRNA-seq) techniques have allowed us to explore the sparsely activated engram ensembles, enabling access to the molecular mechanisms that underlie experience-dependent memory formation and consolidation. However, the absence of specific and powerful computational methods to detect memory-related genes (modules) and their regulatory relationships in the sc/snRNA-seq datasets has strictly limited the analysis of underlying mechanisms and memory coding principles in mammalian brains. Here, we present a deep-learning method named SCENTBOX, to detect memory-related gene modules and causal regulatory relationships among themfromsc/snRNA-seq datasets. SCENTBOX first constructs codifferential expression gene network (CEGN) from case versus control sc/snRNA-seq datasets. It then detects the highly correlated modules of differential expression genes (DEGs) in CEGN. The deep network embedding and attention-based convolutional neural network strategies are employed to precisely detect regulatory relationships among DEG genes in a module. We applied them on scRNA-seq datasets of TRAP; Ai14 mouse neurons with fear memory and detected not only known memory-related genes, but also the modules and potential causal regulations. Our results provided novel regulations within an interesting module, including Arc, Bdnf, Creb, Dusp1, Rgs4, and Btg2. Overall, our methods provide a general computational tool for processing sc/snRNA-seq data from case versus control studie and a systematic investigation of fear-memory-related gene modules.

Keywords: sc/snRNA-seq, memory formation, deep learning, gene module, causal inference

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2410 Improving the Training for Civil Engineers by Introducing Virtual Reality Technique

Authors: Manar Al-Ateeq

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The building construction industry plays a major role in the economy of the word and the state of Kuwait. This paper evaluates existing new civil site engineers, describes a new system for improvement and insures the importance of prequalifying and developing for new engineers. In order to have a strong base in engineering, educational institutes and workplaces should be responsible to continuously train engineers and update them with new methods and techniques in engineering. As to achieve that, school of engineering should constantly update computational resources to be used in the professions. A survey was prepared for graduated Engineers based on stated objectives to understand the status of graduate engineers in both the public and private sector. Interviews were made with different sectors in Kuwait, and several visits were made to different training centers within different workplaces in Kuwait to evaluate training process and try to improve it. Virtual Reality (VR) technology could be applied as a complement to three-dimensional (3D) modeling, leading to better communication whether in job training, in education or in professional practice. Techniques of 3D modeling and VR can be applied to develop the models related to the construction process. The 3D models can support rehabilitation design as it can be considered as a great tool for monitoring failure and defaults in structures; also it can support decisions based on the visual analyses of alternative solutions. Therefore, teaching computer-aided design (CAD) and VR techniques in school will help engineering students in order to prepare them to site work and also will assist them to consider these technologies as important supports in their later professional practice. This teaching technique will show how the construction works developed, allow the visual simulation of progression of each type of work and help them to know more about the necessary equipment needed for tasks and how it works on site.

Keywords: three dimensional modeling (3DM), civil engineers (CE), professional practice (PP), virtual reality (VR)

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2409 Testing Nitrogen and Iron Based Compounds as an Environmentally Safer Alternative to Control Broadleaf Weeds in Turf

Authors: Simran Gill, Samuel Bartels

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Turfgrass is an important component of urban and rural lawns and landscapes. However, broadleaf weeds such as dandelions (Taraxacum officinale) and white clovers (Trifolium repens) pose major challenges to the health and aesthetics of turfgrass fields. Chemical weed control methods, such as 2,4-D weedicides, have been widely deployed; however, their safety and environmental impacts are often debated. Alternative, environmentally friendly control methods have been considered, but experimental tests for their effectiveness have been limited. This study investigates the use and effectiveness of nitrogen and iron compounds as nutrient management methods of weed control. In a two-phase experiment, the first conducted on a blend of cool season turfgrasses in plastic containers, the blend included Perennial ryegrass (Lolium perenne), Kentucky bluegrass (Poa pratensis) and Creeping red fescue (Festuca rubra) grown under controlled conditions in the greenhouse, involved the application of different combinations of nitrogen (urea and ammonium sulphate) and iron (chelated iron and iron sulphate) compounds and their combinations (urea × chelated iron, urea × iron sulphate, ammonium sulphate × chelated iron, ammonium sulphate × iron sulphate) contrasted with chemical 2, 4-D weedicide and a control (no application) treatment. There were three replicates of each of the treatments, resulting in a total of 30 treatment combinations. The parameters assessed during weekly data collection included a visual quality rating of weeds (nominal scale of 0-9), number of leaves, longest leaf span, number of weeds, chlorophyll fluorescence of grass, the visual quality rating of grass (0-9), and the weight of dried grass clippings. The results drawn from the experiment conducted over the period of 12 weeks, with three applications each at an interval of every 4 weeks, stated that the combination of ammonium sulphate and iron sulphate appeared to be most effective in halting the growth and establishment of dandelions and clovers while it also improved turf health. The second phase of the experiment, which involved the ammonium sulphate × iron sulphate, weedicide, and control treatments, was conducted outdoors on already established perennial turf with weeds under natural field conditions. After 12 weeks of observation, the results were comparable among the treatments in terms of weed control, but the ammonium sulphate × iron sulphate treatment fared much better in terms of the improved visual quality of the turf and other quality ratings. Preliminary results from these experiments thus suggest that nutrient management based on nitrogen and iron compounds could be a useful environmentally friendly alternative for controlling broadleaf weeds and improving the health and quality of turfgrass.

Keywords: broadleaf weeds, nitrogen, iron, turfgrass

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2408 Molecular Mechanisms of Lipid Metabolism and Obesity Modulation by Caspase-1/11 and nlrp3 Inflammasome in Mice

Authors: Lívia Pimentel Sant'ana Dourado, Raquel Das Neves Almeida, Luís Henrique Costa Corrêa Neto, Nayara Soares, Kelly Grace Magalhães

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Introduction: Obesity and high-fat diet intake have a crucial impact on immune cells and inflammatory profile, highlighting an emerging realization that obesity is an inflammatory disease. In the present work, we aimed to characterize the role of caspase-1/11 and NLRP3 inflammasome in the establishment of mice obesity and modulation of inflammatory lipid metabolism induced by high fat diet intake. Methods and results: Wild type, caspase-1/11 and NLRP3 knockout mice were fed with standard fat diet (SFD) or high fat diet (HFD) for 90 days. The weight of animals was measured weekly to monitor the weight gain. After 90 days, the blood, peritoneal lavage cells, heart and liver were collected from mice studied here. Cytokines were measured in serum by ELISA and analyzed in spectrophotometry. Lipid antigen presentation molecule CD1d expression, reactive oxygen species (ROS) generation and lipid droplets biogenesis were analyzed in cells from mice peritoneal cavity by flow cytometry. Liver histopathology was performed for morphological evaluation of the organ. The absence of caspase-1/11, but not NLRP3, in mice fed with HFD favored the mice weight gain, increased liver size, induced development of hepatic steatosis and IL-12 secretion in mice compared to mice fed with SFD. In addition, caspase-1/11 knockout mice fed with HFD presented an increased CD1d molecule expression, as well as higher levels of lipid droplets biogenesis and ROS generation compared to wild type mice also fed with HFD. Conclusion: Our data suggest that caspase-1/11 knockout mice have greater susceptibility to obesity as well as increased activation of lipid metabolism and inflammatory markers.

Keywords: caspase 1, caspase 11, inflamassome, obesity, lipids

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2407 Multimodal Analysis of News Magazines' Front-Page Portrayals of the US, Germany, China, and Russia

Authors: Alena Radina

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On the global stage, national image is shaped by historical memory of wars and alliances, government ideology and particularly media stereotypes which represent countries in positive or negative ways. News magazine covers are a key site for national representation. The object of analysis in this paper is the portrayals of the US, Germany, China, and Russia in the front pages and cover stories of “Time”, “Der Spiegel”, “Beijing Review”, and “Expert”. Political comedy helps people learn about current affairs even if politics is not their area of interest, and thus satire indirectly sets the public agenda. Coupled with satirical messages, cover images and the linguistic messages embedded in the covers become persuasive visual and verbal factors, known to drive about 80% of magazine sales. Preliminary analysis identified satirical elements in magazine covers, which are known to influence and frame understandings and attract younger audiences. Multimodal and transnational comparative framing analyses lay the groundwork to investigate why journalists, editors and designers deploy certain frames rather than others. This research investigates to what degree frames used in covers correlate with frames within the cover stories and what these framings can tell us about media professionals’ representations of their own and other nations. The study sample includes 32 covers consisting of two covers representing each of the four chosen countries from the four magazines. The sampling framework considers two time periods to compare countries’ representation with two different presidents, and between men and women when present. The countries selected for analysis represent each category of the international news flows model: the core nations are the US and Germany; China is a semi-peripheral country; and Russia is peripheral. Examining textual and visual design elements on the covers and images in the cover stories reveals not only what editors believe visually attracts the reader’s attention to the magazine but also how the magazines frame and construct national images and national leaders. The cover is the most powerful editorial and design page in a magazine because images incorporate less intrusive framing tools. Thus, covers require less cognitive effort of audiences who may therefore be more likely to accept the visual frame without question. Analysis of design and linguistic elements in magazine covers helps to understand how media outlets shape their audience’s perceptions and how magazines frame global issues. While previous multimodal research of covers has focused mostly on lifestyle magazines or newspapers, this paper examines the power of current affairs magazines’ covers to shape audience perception of national image.

Keywords: framing analysis, magazine covers, multimodality, national image, satire

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2406 A Theoretical Study on Pain Assessment through Human Facial Expresion

Authors: Mrinal Kanti Bhowmik, Debanjana Debnath Jr., Debotosh Bhattacharjee

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A facial expression is undeniably the human manners. It is a significant channel for human communication and can be applied to extract emotional features accurately. People in pain often show variations in facial expressions that are readily observable to others. A core of actions is likely to occur or to increase in intensity when people are in pain. To illustrate the changes in the facial appearance, a system known as Facial Action Coding System (FACS) is pioneered by Ekman and Friesen for human observers. According to Prkachin and Solomon, a set of such actions carries the bulk of information about pain. Thus, the Prkachin and Solomon pain intensity (PSPI) metric is defined. So, it is very important to notice that facial expressions, being a behavioral source in communication media, provide an important opening into the issues of non-verbal communication in pain. People express their pain in many ways, and this pain behavior is the basis on which most inferences about pain are drawn in clinical and research settings. Hence, to understand the roles of different pain behaviors, it is essential to study the properties. For the past several years, the studies are concentrated on the properties of one specific form of pain behavior i.e. facial expression. This paper represents a comprehensive study on pain assessment that can model and estimate the intensity of pain that the patient is suffering. It also reviews the historical background of different pain assessment techniques in the context of painful expressions. Different approaches incorporate FACS from psychological views and a pain intensity score using the PSPI metric in pain estimation. This paper investigates in depth analysis of different approaches used in pain estimation and presents different observations found from each technique. It also offers a brief study on different distinguishing features of real and fake pain. Therefore, the necessity of the study lies in the emerging fields of painful face assessment in clinical settings.

Keywords: facial action coding system (FACS), pain, pain behavior, Prkachin and Solomon pain intensity (PSPI)

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2405 Colloquialism in Audiovisual Translation: English Subtitling of the Lebanese Film Capernaum as a Case Study

Authors: Fatima Saab

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This paper attempts to study colloquialism in audio-visual translation, with particular emphasis given to investigating the difficulties and challenges encountered by subtitlers in translating Lebanese colloquial into English. To achieve the main objectives of this study, ample and thorough cultural and translational analysis of examples drawn from the subtitled movie Capernaum are presented in order to identify the strategies used to overcome cultural barriers and differences and to show the process of decision-making by the translator. Also, special attention is given to explain the technicalities in translating subtitles and how they affect the translation process. The research is a descriptive analytical study whereby the writer sets out empirical observations, consisting of descriptive and analytical examination of the difficulties and problems associated with translating Arabic colloquialisms, specifically Lebanese, into English in the subtitled film, Capernaum. The research methodology utilizes a qualitative approach to group the selected data into the subtitling strategies presented by Gottlieb under the domesticating or foreignizing strategies according to Venuti's Model. It is shown that producing the same meanings to a foreign audience is not an easy task. The background of cultural elements and the stories that make up the history and mindset of the Lebanese and Arabic peoples leads to the use of the transfer and paraphrase methodologies most of the time (81% of the sample used for analysis). The research shows that translating and subtitling colloquialism needs special skills by the translators to overcome the challenges imposed by the limited presentation space as well as cultural differences. Translation of colloquial Arabic/Lebanese can be achieved to a certain extent and delivering the meaning and effect of the source language culture is accomplished in as much as the translator investigates and relates to the target culture.

Keywords: Lebanese colloquial, audio-visual translation, subtitling, Capernaum

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2404 Graphic Narratives: Representations of Refugeehood in the Form of Illustration

Authors: Pauline Blanchet

Abstract:

In a world where images are a prominent part of our daily lives and a way of absorbing information, the analysis of the representation of migration narratives is vital. This thesis raises questions concerning the power of illustrations, drawings and visual culture in order to represent the migration narratives in the age of Instagram. The rise of graphic novels and comics has come about in the last fifteen years, specifically regarding contemporary authors engaging with complex social issues such as migration and refugeehood. Due to this, refugee subjects are often in these narratives, whether they are autobiographical stories or whether the subject is included in the creative process. Growth in discourse around migration has been present in other art forms; in 2018, there has been dedicated exhibitions around migration such as Tania Bruguera at the TATE (2018-2019), ‘Journeys Drawn’ at the House of Illustration (2018-2019) and dedicated film festivals (2018; the Migration Film Festival), which have shown the recent considerations of using the arts as a medium of expression regarding themes of refugeehood and migration. Graphic visuals are fast becoming a key instrument when representing migration, and the central thesis of this paper is to show the strength and limitations of this form as well the methodology used by the actors in the production process. Recent works which have been released in the last ten years have not being analysed in the same context as previous graphic novels such as Palestine and Persepolis. While a lot of research has been done on the mass media portrayals of refugees in photography and journalism, there is a lack of literature on the representation with illustrations. There is little research about the accessibility of graphic novels such as where they can be found and what the intentions are when writing the novels. It is interesting to see why these authors, NGOs, and curators have decided to highlight these migrant narratives in a time when the mainstream media has done extensive coverage on the ‘refugee crisis’. Using primary data by doing one on one interviews with artists, curators, and NGOs, this paper investigates the efficiency of graphic novels for depicting refugee stories as a viable alternative to other mass medium forms. The paper has been divided into two distinct sections. The first part is concerned with the form of the comic itself and how it either limits or strengthens the representation of migrant narratives. This will involve analysing the layered and complex forms that comics allow such as multimedia pieces, use of photography and forms of symbolism. It will also show how the illustration allows for anonymity of refugees, the empathetic aspect of the form and how the history of the graphic novel form has allowed space for positive representations of women in the last decade. The second section will analyse the creative and methodological process which takes place by the actors and their involvement with the production of the works.

Keywords: graphic novel, refugee, communication, media, migration

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2403 Effect of Xenobiotic Bioactive Compounds from Grape Waste on Inflammation and Oxidative Stress in Pigs

Authors: Ionelia Taranu, Gina Cecilia Pistol, Mihai Alexandru Gras, Mihai Laurentiu Palade, Mariana Stancu, Veronica Sanda Chedea

Abstract:

In the last decade bioactive compounds from grape waste are investigated as new therapeutic agents for the inhibition of carcinogenesis and other diseases. The objective of this study was to characterize several bioactive compounds (polyphenols and polyunsaturated fatty acids) of a dried grape pomace (GP) derived from a Romanian winery and further to evaluate their effect on inflammation and oxidative markers in liver of pig used as animal model. The total polyphenol concentration of pomace was 36.2g gallic acid equiv /100g. The pomace was rich in polyphenols from the flavonoids group, the main class being flavanols (epicatechins, catechin, epigallocatechin, procyanidins) and antocyanins (Malvidin 3-O-glucoside). The highest concentration was recorded for epicatechin (51.96g/100g) and procyanidin dimer (22.79g/100g). A high concentration of total polyunsaturated fatty acids (PUFA) especially ω-6 fatty acids (59.82 g/100g fat) was found in grape pomace. 20 crossbred TOPIG hybrid fattening pigs were randomly assigned (n = 10) to two experimental treatments: a normal diet (control group) and a diet included 5% grape pomace (GP group) for 24 days. The GP diet lowered the gene expression and protein concentration of IL-1β, IL-8, TNF-α and IFN-γ cytokines in liver suggesting an anti-inflammatory effect of GP diet. Concentration of hepatic TBARS also decreased, but the total antioxidant capacity (liver TEAC) and activity and gene expression of antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) did not differ between the GP and control diet. The results showed that GP diet exerted an anti-inflammatory effect, but the 5% dietary inclusion modulated only partially the oxidative stress.

Keywords: animal model, inflammation, grape waste, immune organs

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2402 Developing Scaffolds for Tissue Regeneration using Low Temperature Plasma (LTP)

Authors: Komal Vig

Abstract:

Cardiovascular disease (CVD)-related deaths occur in 17.3 million people globally each year, accounting for 30% of all deaths worldwide, with a predicted annual incidence of deaths to reach 23.3 million globally by 2030. Autologous bypass grafts remain an important therapeutic option for the treatment of CVD, but the poor quality of the donor patient’s blood vessels, the invasiveness of the resection surgery, and postoperative movement restrictions create issues. The present study is aimed to improve the endothelialization of intimal surface of graft by using low temperature plasma (LTP) to increase the cell attachment and proliferation. Polytetrafluoroethylene (PTFE) was treated with LTP. Air was used as the feed-gas, and the pressure in the plasma chamber was kept at 800 mTorr. Scaffolds were also modified with gelatin and collagen by dipping method. Human umbilical vein endothelial cells (HUVEC) were plated on the developed scaffolds, and cell proliferation was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by microscopy. mRNA expressions levels of different cell markers were investigated using quantitative real-time PCR (qPCR). XPS confirmed the introduction of oxygenated functionalities from LTP. HUVEC cells showed 80% seeding efficiency on the scaffold. Microscopic and MTT assays indicated increase in cell viability in LTP treated scaffolds, especially when treated with gelatin or collagen, compared to untreated scaffolds. Gene expression studies shows enhanced expression of cell adhesion marker Integrin- α 5 gene after LTP treatment. LTP treated scaffolds exhibited better cell proliferation and viability compared to untreated scaffolds. Protein treatment of scaffold increased cell proliferation. Based on our initial results, more scaffolds alternatives will be developed and investigated for cell growth and vascularization studies. Acknowledgments: This work is supported by the NSF EPSCoR RII-Track-1 Cooperative Agreement OIA-2148653.

Keywords: LTP, HUVEC cells, vascular graft, endothelialization

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2401 Magnetic Properties of Nickel Oxide Nanoparticles in Superparamagnetic State

Authors: Navneet Kaur, S. D. Tiwari

Abstract:

Superparamagnetism is an interesting phenomenon and observed in small particles of magnetic materials. It arises due to a reduction in particle size. In the superparamagnetic state, as the thermal energy overcomes magnetic anisotropy energy, the magnetic moment vector of particles flip their magnetization direction between states of minimum energy. Superparamagnetic nanoparticles have been attracting the researchers due to many applications such as information storage, magnetic resonance imaging, biomedical applications, and sensors. For information storage, thermal fluctuations lead to loss of data. So that nanoparticles should have high blocking temperature. And to achieve this, nanoparticles should have a higher magnetic moment and magnetic anisotropy constant. In this work, the magnetic anisotropy constant of the antiferromagnetic nanoparticles system is determined. Magnetic studies on nanoparticles of NiO (nickel oxide) are reported well. This antiferromagnetic nanoparticle system has high blocking temperature and magnetic anisotropy constant of order 105 J/m3. The magnetic study of NiO nanoparticles in the superparamagnetic region is presented. NiO particles of two different sizes, i.e., 6 and 8 nm, are synthesized using the chemical route. These particles are characterized by an x-ray diffractometer, transmission electron microscope, and superconducting quantum interference device magnetometry. The magnetization vs. applied magnetic field and temperature data for both samples confirm their superparamagnetic nature. The blocking temperature for 6 and 8 nm particles is found to be 200 and 172 K, respectively. Magnetization vs. applied magnetic field data of NiO is fitted to an appropriate magnetic expression using a non-linear least square fit method. The role of particle size distribution and magnetic anisotropy is taken in to account in magnetization expression. The source code is written in Python programming language. This fitting provides us the magnetic anisotropy constant for NiO and other magnetic fit parameters. The particle size distribution estimated matches well with the transmission electron micrograph. The value of magnetic anisotropy constants for 6 and 8 nm particles is found to be 1.42 X 105 and 1.20 X 105 J/m3, respectively. The obtained magnetic fit parameters are verified using the Neel model. It is concluded that the effect of magnetic anisotropy should not be ignored while studying the magnetization process of nanoparticles.

Keywords: anisotropy, superparamagnetic, nanoparticle, magnetization

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2400 Integrated Life Skill Training and Executive Function Strategies in Children with Autism Spectrum Disorder in Qatar: A Study Protocol for a Randomized Controlled Trial

Authors: Bara M Yousef, Naresh B Raj, Nadiah W Arfah, Brightlin N Dhas

Abstract:

Background: Executive function (EF) impairment is common in children with autism spectrum disorder (ASD). EF strategies are considered effective in improving the therapeutic outcomes of children with ASD. Aims: This study primarily aims to explore whether integrating EF strategies combined with regular occupational therapy intervention is more effective in improving daily life skills (DLS) and sensory integration/processing (SI/SP) skills than regular occupational therapy alone in children with ASD and secondarily aims to assess treatment outcomes on improving visual motor integration (VMI) skills. Procedures: A total of 92 children with ASD will be recruited and, following baseline assessments, randomly assigned to the treatment group (45-min once weekly individual occupational therapy plus EF strategies) and control group (45-min once weekly individual therapy sessions alone). Results and Outcomes: All children will be evaluated systematically by assessing SI/SP, DLS, and VMI, skills at baseline, 7 weeks, and 14 weeks of treatment. Data will be analyzed using ANCOVA and T-test. Conclusions and Implications: This single-blind, randomized controlled trial will provide empirical evidence for the effectiveness of EF strategies when combined with regular occupational therapy programs. Based on trial results, EF strategies could be recommended in multidisciplinary programs for children with ASD. Trial Registration: The trial has been registered in the clinicaltrail.gov for a registry, protocol ID: MRC-01-22-509 ClinicalTrials.gov Identifier: NCT05829577, registered 25th April 2023

Keywords: autism spectrum disorder, executive function strategies, daily life skills, sensory integration/processing, visual motor integration, occupational therapy, effectiveness

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2399 Identification of Hub Genes in the Development of Atherosclerosis

Authors: Jie Lin, Yiwen Pan, Li Zhang, Zhangyong Xia

Abstract:

Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of lipids, immune cells, and extracellular matrix in the arterial walls. This pathological process can lead to the formation of plaques that can obstruct blood flow and trigger various cardiovascular diseases such as heart attack and stroke. The underlying molecular mechanisms still remain unclear, although many studies revealed the dysfunction of endothelial cells, recruitment and activation of monocytes and macrophages, and the production of pro-inflammatory cytokines and chemokines in atherosclerosis. This study aimed to identify hub genes involved in the progression of atherosclerosis and to analyze their biological function in silico, thereby enhancing our understanding of the disease’s molecular mechanisms. Through the analysis of microarray data, we examined the gene expression in media and neo-intima from plaques, as well as distant macroscopically intact tissue, across a cohort of 32 hypertensive patients. Initially, 112 differentially expressed genes (DEGs) were identified. Subsequent immune infiltration analysis indicated a predominant presence of 27 immune cell types in the atherosclerosis group, particularly noting an increase in monocytes and macrophages. In the Weighted gene co-expression network analysis (WGCNA), 10 modules with a minimum of 30 genes were defined as key modules, with blue, dark, Oliver green and sky-blue modules being the most significant. These modules corresponded respectively to monocyte, activated B cell, and activated CD4 T cell gene patterns, revealing a strong morphological-genetic correlation. From these three gene patterns (modules morphology), a total of 2509 key genes (Gene Significance >0.2, module membership>0.8) were extracted. Six hub genes (CD36, DPP4, HMOX1, PLA2G7, PLN2, and ACADL) were then identified by intersecting 2509 key genes, 102 DEGs with lipid-related genes from the Genecard database. The bio-functional analysis of six hub genes was estimated by a robust classifier with an area under the curve (AUC) of 0.873 in the ROC plot, indicating excellent efficacy in differentiating between the disease and control group. Moreover, PCA visualization demonstrated clear separation between the groups based on these six hub genes, suggesting their potential utility as classification features in predictive models. Protein-protein interaction (PPI) analysis highlighted DPP4 as the most interconnected gene. Within the constructed key gene-drug network, 462 drugs were predicted, with ursodeoxycholic acid (UDCA) being identified as a potential therapeutic agent for modulating DPP4 expression. In summary, our study identified critical hub genes implicated in the progression of atherosclerosis through comprehensive bioinformatic analyses. These findings not only advance our understanding of the disease but also pave the way for applying similar analytical frameworks and predictive models to other diseases, thereby broadening the potential for clinical applications and therapeutic discoveries.

Keywords: atherosclerosis, hub genes, drug prediction, bioinformatics

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2398 The Effectiveness of Laser In situ Keratomileusis for Correction Various Types of Refractive Anomalies

Authors: Yuliya Markava

Abstract:

The laser in situ keratomileusis (LASIK) is widely common surgical procedure, which has become an alternative for patients who are not satisfied with the performance of other correction methods. A high level of patient satisfaction functional outcomes after refractive surgery confirms the high reliability and safety of LASIK and provides a significant improvement in the quality of life and social adaptation. Purpose: To perform clinical analysis of the results of correction made to the excimer laser system SCHWIND AMARIS 500E in patients with different types of refractive anomalies. Materials and Methods: This was a retrospective analysis of 1581 operations (812 patients): 413 males (50.86%) and 399 females (49.14%) at the age from 18 to 47 years with different types of ametropia. All operations were performed on excimer laser SCHWIND AMARIS 500E in the LASIK procedure. Formation of the corneal flap was made by mechanical microkeratome SCHWIND. Results: Analyzing the structure of refractive anomalies: The largest number of interventions accounted for myopia: 1505 eyes (95.2%), of which about a low myopia: 706 eyes (44.7%), moderate myopia: 562 eyes (35.5 %), high myopia: eyes 217 (13.7%) and supermyopia: 20 eyes (1.3%). Hyperopia was 0.7% (11 eyes), mixed astigmatism: 4.1% (65 eyes). The efficiency was 80% (in patients with supermyopia) to 91.6% and 95.4% (in groups with myopia low and moderate, respectively). Uncorrected visual acuity average values before and after laser operation was in groups: a low myopia 0.18 (up 0.05 to 0.31) and 0.80 (up 0.60 to 1.0); moderate myopia 0.08 (up 0.03 to 0.13) and 0.87 ( up 0.74 to 1.0); high myopia 0.05 (up 0.02 to 0.08) and 0.83 (up 0.66 to 1.0); supermyopia 0.03 (up 0.02 to 0.04) and 0.59 ( up 0.34 to 0.84); hyperopia 0.27 (up 0.16 to 0.38) and 0.57 (up 0.27 to 0.87); mixed astigmatism of 0.35 (up 0.19 to 0.51) and 0.69 (up 0.44 to 0.94). In all cases, after LASIK indicators uncorrected visual acuity significantly increased. Reoperation was 4.43%. Significance: Clinical results of refractive surgery at the excimer laser system SCHWIND AMARIS 500E in different ametropia correction is characterized by high efficiency.

Keywords: effectiveness of laser correction, LASIK, refractive anomalies, surgical treatment

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2397 The Process of Irony Comprehension in Young Children: Evidence from Monolingual and Bilingual Preschoolers

Authors: Natalia Banasik

Abstract:

Comprehension of verbal irony is an example of pragmatic competence in understanding figurative language. The knowledge of how it develops may shed new light on the understanding of social and communicative competence that is crucial for one's effective functioning in the society. Researchers agree it is a competence that develops late in a child’s development. One of the abilities that seems crucial for irony comprehension is theory of mind (ToM), that is the ability to understand that others may have beliefs, desires and intentions different from one’s own. Although both theory of mind and irony comprehension require the ability to understand the figurative use of the false description of the reality, the exact relationship between them is still unknown. Also, even though irony comprehension in children has been studied for over thirty years, the results of the studies are inconsistent as to the age when this competence are acquired. The presented study aimed to answer questions about the developmental trajectories of irony comprehension and ascribing function to ironic utterances by preschool children. Specifically, we were interested in how it is related to the development of ToM and how comprehension of the function of irony changes with age. Data was collected from over 150 monolingual, Polish-speaking children and (so far) thirty bilingual children speaking Polish and English who live in the US. Four-, five- and six-year-olds were presented with a story comprehension task in the form of audio and visual stimuli programmed in the E-prime software (pre-recorded narrated stories, some of which included ironic utterances, and pictures accompanying the stories displayed on a touch screen). Following the presentation, the children were then asked to answer a series of questions. The questions checked the children’s understanding of the intended utterance meaning, evaluation of the degree to which it was funny and evaluation of how nice the speaker was. The children responded by touching the screen, which made it possible to measure reaction times. Additionally, the children were asked to explain why the speaker had uttered the ironic statement. Both quantitive and qualitative analyses were applied. The results of our study indicate that for irony recognition there is a significant difference among the three age groups, but what is new is that children as young as four do understand the real meaning behind the ironic statement as long as the utterance is not grammtically or lexically complex also, there is a clear correlation of ToM and irony comprehension. Although four-year olds and six-year olds understand the real meaning of the ironic utterance, it is not earlier than at the age of six when children start to explain the reason of using this marked form of expression. They talk about the speaker's intention to tell a joke, be funny, or to protect the listener's emotions. There are also some metalinguistic references, such as "mommy sometimes says things that don't make sense and this is called a metaphor".

Keywords: child's pragmatics, figurative speech, irony comprehension in children, theory of mind and irony

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2396 The Effect of Common Daily Schedule on the Human Circadian Rhythms during the Polar Day on Svalbard: Field Study

Authors: Kamila Weissova, Jitka Skrabalova, Katerina Skalova, Jana Koprivova, Zdenka Bendova

Abstract:

Any Arctic visitor has to deal with extreme conditions, including constant light during the summer season or constant darkness during winter time. Light/dark cycle is the most powerful synchronizing signal for biological clock and the absence of daily dark period during the polar day can significantly alter the functional state of the internal clock. However, the inner clock can be synchronized by other zeitgebers such as physical activity, food intake or social interactions. Here, we investigated the effect of polar day on circadian clock of 10 researchers attending the polar base station in the Svalbard region during July. The data obtained on Svalbard were compared with the data obtained before the researchers left for the expedition (in the Czech Republic). To determine the state of circadian clock we used wrist actigraphy followed by sleep diaries, saliva, and buccal mucosa samples, both collected every 4 hours during 24h-interval to detect melatonin by radioimmunoassay and clock gene (PER1, BMAL1, NR1D1, DBP) mRNA levels by RT-qPCR. The clock gene expression was analyzed using cosinor analysis. From our results, it is apparent that the constant sunlight delayed melatonin onset and postponed the physical activity in the same order. Nevertheless, the clock gene expression displayed higher amplitude on Svalbard compared to the amplitude detected in the Czech Republic. These results have suggested that the common daily schedule at the Svalbard expedition can strengthen circadian rhythm in the environment that is lacking light/dark cycle. In conclusion, the constant sunlight delays melatonin onset, but it still maintains its rhythmic secretion. The effect of constant sunlight on circadian clock can be minimalized by common daily scheduled activity.

Keywords: actighraph, clock genes, human, melatonin, polar day

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2395 Artificial Cells Capable of Communication by Using Polymer Hydrogel

Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

Abstract:

The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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2394 [Keynote Talk]: Bioactive Cyclic Dipeptides of Microbial Origin in Discovery of Cytokine Inhibitors

Authors: Sajeli A. Begum, Ameer Basha, Kirti Hira, Rukaiyya Khan

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Cyclic dipeptides are simple diketopiperazine derivatives being investigated by several scientists for their biological effects which include anticancer, antimicrobial, haematological, anticonvulsant, immunomodulatory effect, etc. They are potentially active microbial metabolites having been synthesized too, for developing into drug candidates. Cultures of Pseudomonas species have earlier been reported to produce cyclic dipeptides, helping in quorum sensing signals and bacterial–host colonization phenomena during infections, causing cell anti-proliferation and immunosuppression. Fluorescing Pseudomonas species have been identified to secrete lipid derivatives, peptides, pyrroles, phenazines, indoles, aminoacids, pterines, pseudomonic acids and some antibiotics. In the present work, results of investigation on the cyclic dipeptide metabolites secreted by the culture broth of Pseudomonas species as potent pro-inflammatory cytokine inhibitors are discussed. The bacterial strain was isolated from the rhizospheric soil of groundnut crop and identified as Pseudomonas aeruginosa by 16S rDNA sequence (GenBank Accession No. KT625586). Culture broth of this strain was prepared by inoculating into King’s B broth and incubating at 30 ºC for 7 days. The ethyl acetate extract of culture broth was prepared and lyophilized to get a dry residue (EEPA). Lipopolysaccharide (LPS)-induced ELISA assay proved the inhibition of tumor necrosis factor-alpha (TNF-α) secretion in culture supernatant of RAW 264.7 cells by EEPA (IC50 38.8 μg/mL). The effect of oral administration of EEPA on plasma TNF-α level in rats was tested by ELISA kit. The LPS mediated plasma TNF-α level was reduced to 45% with 125 mg/kg dose of EEPA. Isolation of the chemical constituents of EEPA through column chromatography yielded ten cyclic dipeptides, which were characterized using nuclear magnetic resonance and mass spectroscopic techniques. These cyclic dipeptides are biosynthesized in microorganisms by multifunctional assembly of non-ribosomal peptide synthases and cyclic dipeptide synthase. Cyclo (Gly-L-Pro) was found to be more potentially (IC50 value 4.5 μg/mL) inhibiting TNF-α production followed by cyclo (trans-4-hydroxy-L-Pro-L-Phe) (IC50 value 14.2 μg/mL) and the effect was equal to that of standard immunosuppressant drug, prednisolone. Further, the effect was analyzed by determining mRNA expression of TNF-α in LPS-stimulated RAW 264.7 macrophages using quantitative real-time reverse transcription polymerase chain reaction. EEPA and isolated cyclic dipeptides demonstrated diminution of TNF-α mRNA expression levels in a dose-dependent manner under the tested conditions. Also, they were found to control the expression of other pro-inflammatory cytokines like IL-1β and IL-6, when tested through their mRNA expression levels in LPS-stimulated RAW 264.7 macrophages under LPS-stimulated conditions. In addition, significant inhibition effect was found on Nitric oxide production. Further all the compounds exhibited weak toxicity to LPS-induced RAW 264.7 cells. Thus the outcome of the study disclosed the effectiveness of EEPA and the isolated cyclic dipeptides in down-regulating key cytokines involved in pathophysiology of autoimmune diseases.In another study led by the investigators, microbial cyclic dipeptides were found to exhibit excellent antimicrobial effect against Fusarium moniliforme which is an important causative agent of Sorghum grain mold disease. Thus, cyclic dipeptides are emerging small molecular drug candidates for various autoimmune diseases.

Keywords: cyclic dipeptides, cytokines, Fusarium moniliforme, Pseudomonas, TNF-alpha

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2393 Upper Bound of the Generalized P-Value for the Difference between Two Future Population Means

Authors: Rada Somkhuean, Sa-aat Niwitpong, Suparat Niwitpong

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This paper presents the generalized p-values for testing the difference between two future population means when the variances are unknown, in both cases for when the variances are equal and unequal. We also derive a closed form expression of the upper bound of the proposed generalized p-value.

Keywords: generalized p-value, two future population means, upper bound, variances

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2392 Gene Expression Analysis for Corals / Zooxanthellae under High Seawater Temperature Stress

Authors: Haruka Ito, Toru Maruyama, Michihiro Ito, Chuya Shinzato, Hiroyuki Fujimura, Yoshikatsu Nakano, Shoichiro Suda, Sachiyo Aburatani, Haruko Takeyama

Abstract:

Clarifying symbiotic relationships is one of the most important theme for understanding the marine eco-system. Coral reef has been regarded as an important environmental resource. Coral holobiont composed by coral, symbiotic microalgae zooxanthellae, and bacteria have complexed relationship. Zooxanthellae mainly supply organic matter to the host corals through their photosynthetic activity. The symbiotic relationship is indispensable for corals but may easily collapses due to the rise of seawater temperature. However, the molecular mechanism how seawater temperature influences their relationships still remain unclear. In this study, the transcriptomic analysis has applied to elucidate the coral-zooxanthellae relationships under high seawater temperature stress. To observe reactions of corals and zooxanthellae against the rise of seawater temperature, meta-gene expression in coral have been analyzed. The branches from six different colonies of a stony coral, Acropora tenuis, were sampled at nine times by 2016 at two locations, Ishikawabaru and South of Sesoko Island, Okinawa, Japan. The mRNAs extracted from the branches including zooxanthellae were sequenced by illumina HiSeq. Gene Set Enrichment Analysis (GSEA) based on hyper geometric distribution was performed. The seawater temperature at 2016 summer was unusually high, which was caused by El Niño event, and the number of zooxanthellae in coral was decreased in August. GSEA derived the several specific genes expressed in A. tenuis under heat stress conditions. The upregulated genes under heat stress highly related with infection immunity. The downregulated genes significantly contained cell cycle related genes. Thu, it is considered that heat stress cause disorder in cell metabolism of A. tenuis, resulting in serious influence to coral holobiont.

Keywords: coral, symbiosis, thermal stress response, transcriptome analysis

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2391 Multi-Sensor Image Fusion for Visible and Infrared Thermal Images

Authors: Amit Kumar Happy

Abstract:

This paper is motivated by the importance of multi-sensor image fusion with a specific focus on infrared (IR) and visual image (VI) fusion for various applications, including military reconnaissance. Image fusion can be defined as the process of combining two or more source images into a single composite image with extended information content that improves visual perception or feature extraction. These images can be from different modalities like visible camera & IR thermal imager. While visible images are captured by reflected radiations in the visible spectrum, the thermal images are formed from thermal radiation (infrared) that may be reflected or self-emitted. A digital color camera captures the visible source image, and a thermal infrared camera acquires the thermal source image. In this paper, some image fusion algorithms based upon multi-scale transform (MST) and region-based selection rule with consistency verification have been proposed and presented. This research includes the implementation of the proposed image fusion algorithm in MATLAB along with a comparative analysis to decide the optimum number of levels for MST and the coefficient fusion rule. The results are presented, and several commonly used evaluation metrics are used to assess the suggested method's validity. Experiments show that the proposed approach is capable of producing good fusion results. While deploying our image fusion algorithm approaches, we observe several challenges from the popular image fusion methods. While high computational cost and complex processing steps of image fusion algorithms provide accurate fused results, they also make it hard to become deployed in systems and applications that require a real-time operation, high flexibility, and low computation ability. So, the methods presented in this paper offer good results with minimum time complexity.

Keywords: image fusion, IR thermal imager, multi-sensor, multi-scale transform

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