Search results for: target specificity
2382 Magnetic Silica Nanoparticles as Viable Support for the Immobilization of Oxidative Enzymes
Authors: Y. Moldes-Diz, M. Gamallo, G. Eibes, C. Vazquez-Vazquez, G. Feijoo, J. M. Lema, M. T. Moreira
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Laccases (benzenediol oxygen oxidoreductases, EC 1.10.3.2) are excellent biocatalysts for biotechnological and environmental applications because of their high activity, selectivity, and specificity. Specifically, these characteristics allow them to perform the oxidation of recalcitrant compounds with simple requirements for the catalysis (presence of molecular oxygen). Nevertheless, the low stability under unfavorable conditions (pH, inactivating agents or temperature) and high production costs still limits their use for practical applications. Immobilization of enzymes has proven particularly valuable to avoid some of the aforementioned drawbacks. Magnetic nanoparticles (MNPs) have received increasing attention as carriers for enzyme immobilization since they can potentially provide an easy recovery of the biocatalyst from the reaction medium under an external magnetic field. In the present work, silica-coated magnetic nanoparticles (Fe3O4@SiO2) were prepared, characterized and used for laccase immobilization by covalent binding. The synthesis of Fe3O4@SiO2 was performed in a two-step procedure: co-precipitation and reverse microemulsion. The influence of immobilization conditions: concentrations of the functionalization agent (3-aminopropyl-triethoxy-silane) and the cross-linker (glutaraldehyde) as well as the influence of pH, T or inactivating agents were evaluated. In general, immobilized laccase showed superior stability compared to that of free enzyme. The reusability of the biocatalyst was demonstrated in successive batch reactions, where enzyme activity was maintained above 65% after 8 cycles of oxidation of the substrate 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate).Keywords: silica-coated magnetic nanoparticles, laccase, immobilization, regeneration
Procedia PDF Downloads 2192381 Integrating Cost-Benefit Assessment and Contract Design to Support Industrial Symbiosis Deployment
Authors: Robin Molinier
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Industrial symbiosis (I.S) is the realization of Industrial Ecology (I.E) principles in production systems in function. I.S consists in the use of waste materials, fatal energy, recirculated utilities and infrastructure/service sharing as resources for production. Environmental benefits can be achieved from resource conservation but economic profitability is required by the participating actors. I.S indeed involves several actors with their own objectives and resources so that each one must be satisfied by ex-ante arrangements to commit toward I.S execution (investments and transactions). Following the Resource-Based View of transactions we build a modular framework to assess global I.S profitability and to specify each actor’s contributions to costs and benefits in line with their resource endowments and performance requirements formulations. I.S projects specificities implied by the need for customization (asset specificity, non-homogeneity) induce the use of long-term contracts for transactions following Transaction costs economics arguments. Thus we propose first a taxonomy of costs and value drivers for I.S and an assignment to each actor of I.S specific risks that we identified as load profiles mismatch, quality problems and value fluctuations. Then appropriate contractual guidelines (pricing, cost sharing and warranties) that support mutual profitability are derived from the detailed identification of contributions by the cost-benefits model. This analytical framework helps identifying what points to focus on when bargaining over contracting for transactions and investments. Our methodology is applied to I.S archetypes raised from a literature survey on eco-industrial parks initiatives and practitioners interviews.Keywords: contracts, cost-benefit analysis, industrial symbiosis, risks
Procedia PDF Downloads 3402380 Review on PETG Material Parts Made Using Fused Deposition Modeling
Authors: Dhval Chauhan, Mahesh Chudasama
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This study has been undertaken to give a review of Polyethylene Terephthalate Glycol (PETG) material used in Fused Deposition Modelling (FDM). This paper offers a review of the existing literature on polyethylene terephthalate glycol (PETG) material, the objective of the paper is to providing guidance on different process parameters that can be used to improve the strength of the part by performing various testing like tensile, compressive, flexural, etc. This work is target to find new paths that can be used for further development of the use of fiber reinforcement in PETG material.Keywords: PETG, FDM, tensile strength, flexural strength, fiber reinforcement
Procedia PDF Downloads 1952379 Programmed Cell Death in Datura and Defensive Plant Response toward Tomato Mosaic Virus
Authors: Asma Alhuqail, Nagwa Aref
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Programmed cell death resembles a real nature active defense in Datura metel against TMV after three days of virus infection. Physiological plant response was assessed for asymptomatic healthy and symptomatic infected detached leaves. The results indicated H2O2 and Chlorophyll-a as the most potential parameters. Chlorophyll-a was considered the only significant predictor variant for the H2O2 dependent variant with a P value of 0.001 and R-square of 0.900. The plant immune response was measured within three days of virus infection using the cutoff value of H2O2 (61.095 lmol/100 mg) and (63.201 units) for the tail moment in the Comet Assay. Their percentage changes were 255.12% and 522.40% respectively which reflects the stress of virus infection in the plant. Moreover, H2O2 showed 100% specificity and sensitivity in the symptomatic infected group using the receiver-operating characteristic (ROC). All tested parameters in the symptomatic infected group had significant correlations with twenty-five positive and thirty-one negative correlations where the P value was <0.05 and 0.01. Chlorophyll-a parameter had a crucial role of highly significant correlation between total protein and salicylic acid. Contrarily, this correlation with tail moment unit was (r = _0.930, P <0.01) where the P value was < 0.01. The strongest significant negative correlation was between Chlorophyll-a and H2O2 at P < 0.01, while moderate negative significant correlation was seen for Chlorophyll-b where the P value < 0.05. The present study discloses the secret of the three days of rapid transient production of activated oxygen species (AOS) that was enough for having potential quantitative physiological parameters for defensive plant response toward the virus.Keywords: programmed cell death, plant–adaptive immune response, hydrogen peroxide (H2O2), physiological parameters
Procedia PDF Downloads 2482378 Identifying and Understand Pragmatic Failures in Portuguese Foreign Language by Chinese Learners in Macau
Authors: Carla Lopes
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It is clear nowadays that the proper performance of different speech acts is one of the most difficult obstacles that a foreign language learner has to overcome to be considered communicatively competent. This communication presents the results of an investigation on the pragmatic performance of Portuguese Language students at the University of Macau. The research discussed herein is based on a survey consisting of fourteen speaking situations to which the participants must respond in writing, and that includes different types of speech acts: apology, response to a compliment, refusal, complaint, disagreement and the understanding of the illocutionary force of indirect speech acts. The responses were classified in a five levels Likert scale (quantified from 1 to 5) according to their suitability for the particular situation. In general terms, we can summarize that about 45% of the respondents' answers were pragmatically competent, 10 % were acceptable and 45 % showed weaknesses at socio-pragmatic competence level. Given that the linguistic deviations were not taken into account, we can conclude that the faults are of cultural origin. It is natural that in the presence of orthogonal cultures, such as Chinese and Portuguese, there are failures of this type, barely solved in the four years of the undergraduate program. The target population, native speakers of Cantonese or Mandarin, make their first contact with the English language before joining the Bachelor of Portuguese Language. An analysis of the socio - pragmatic failures in the respondents’ answers suggests the conclusion that many of them are due to the lack of cultural knowledge. They try to compensate for this either using their native culture or resorting to a Western culture that they consider close to the Portuguese, that is the English or US culture, previously studied, and also widely present in the media and on the internet. This phenomenon, known as 'pragmatic transfer', can result in a linguistic behavior that may be considered inauthentic or pragmatically awkward. The resulting speech act is grammatically correct but is not pragmatically feasible, since it is not suitable to the culture of the target language, either because it does not exist or because the conditions of its use are in fact different. Analysis of the responses also supports the conclusion that these students present large deviations from the expected and stereotyped behavior of Chinese students. We can speculate while this linguistic behavior is the consequence of the Macao globalization that culturally casts the students, makes them more open, and distinguishes them from the typical Chinese students.Keywords: Portuguese foreign language, pragmatic failures, pragmatic transfer, pragmatic competence
Procedia PDF Downloads 2122377 Multi-Channel Information Fusion in C-OTDR Monitoring Systems: Various Approaches to Classify of Targeted Events
Authors: Andrey V. Timofeev
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The paper presents new results concerning selection of optimal information fusion formula for ensembles of C-OTDR channels. The goal of information fusion is to create an integral classificator designed for effective classification of seismoacoustic target events. The LPBoost (LP-β and LP-B variants), the Multiple Kernel Learning, and Weighing of Inversely as Lipschitz Constants (WILC) approaches were compared. The WILC is a brand new approach to optimal fusion of Lipschitz Classifiers Ensembles. Results of practical usage are presented.Keywords: Lipschitz Classifier, classifiers ensembles, LPBoost, C-OTDR systems
Procedia PDF Downloads 4612376 Fabrication of Carbon Nanoparticles and Graphene Using Pulsed Laser Ablation
Authors: Davoud Dorranian, Hajar Sadeghi, Elmira Solati
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Carbon nanostructures in various forms were synthesized using pulsed laser ablation of a graphite target in different liquid environment. The beam of a Q-switched Nd:YAG laser of 1064-nm wavelength at 7-ns pulse width is employed to irradiate the solid target in water, acetone, alcohol, and cetyltrimethylammonium bromide (CTAB). Then the effect of the liquid environment on the characteristic of carbon nanostructures produced by laser ablation was investigated. The optical properties of the carbon nanostructures were examined at room temperature by UV–Vis-NIR spectrophotometer. The crystalline structure of the carbon nanostructures was analyzed by X-ray diffraction (XRD). The morphology of samples was investigated by field emission scanning electron microscope (FE-SEM). Transmission electron microscope (TEM) was employed to investigate the form of carbon nanostructures. Raman spectroscopy was used to determine the quality of carbon nanostructures. Results show that different carbon nanostructures such as nanoparticles and few-layer graphene were formed in various liquid environments. The UV-Vis-NIR absorption spectra of samples reveal that the intensity of absorption peak of nanoparticles in alcohol is higher than the other liquid environments due to the larger number of nanoparticles in this environment. The red shift of the absorption peak of the sample in acetone confirms that produced carbon nanoparticles in this liquid are averagely larger than the other medium. The difference in the intensity and shape of the absorption peak indicated the effect of the liquid environment in producing the nanoparticles. The XRD pattern of the sample in water indicates an amorphous structure due to existence the graphene sheets. X-ray diffraction pattern shows that the degree of crystallinity of sample produced in CTAB is higher than the other liquid environments. Transmission electron microscopy images reveal that the generated carbon materials in water are graphene sheet and in the other liquid environments are graphene sheet and spherical nanostructures. According to the TEM images, we have the larger amount of carbon nanoparticles in the alcohol environment. FE-SEM micrographs indicate that in this liquids sheet like structures are formed however in acetone, produced sheets are adhered and these layers overlap with each other. According to the FE-SEM micrographs, the surface morphology of the sample in CTAB was coarser than that without surfactant. From Raman spectra, it can be concluded the distinct shape, width, and position of the graphene peaks and corresponding graphite source.Keywords: carbon nanostructures, graphene, pulsed laser ablation, graphite
Procedia PDF Downloads 3162375 Partial Knowledge Transfer Between the Source Problem and the Target Problem in Genetic Algorithms
Authors: Terence Soule, Tami Al Ghamdi
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To study how the partial knowledge transfer may affect the Genetic Algorithm (GA) performance, we model the Transfer Learning (TL) process using GA as the model solver. The objective of the TL is to transfer the knowledge from one problem to another related problem. This process imitates how humans think in their daily life. In this paper, we proposed to study a case where the knowledge transferred from the S problem has less information than what the T problem needs. We sampled the transferred population using different strategies of TL. The results showed transfer part of the knowledge is helpful and speeds the GA process of finding a solution to the problem.Keywords: transfer learning, partial transfer, evolutionary computation, genetic algorithm
Procedia PDF Downloads 1322374 Analyzing the Oil and Gas Exploration Opportunities in Poland: Five Prospective Areas Selected and Dedicated to the Tender
Authors: Krystian Wójcik, Sara Wróblewska, Marcin Łojek, Katarzyna Sobień
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Polish Geological Survey selected five of the most prospective areas for oil and gas exploration in Poland. They are dedicated to the 6th international tender round for hydrocarbon concessions, planned in 2022. The main exploration target of these areas is related to conventional and unconventional accumulations of gas and oil in the Carpathian basement, Carpathian Foredeep and Outer Carpathians (Block 413 – 414), as well as in the Carboniferous, Rotliegend, Main Dolomite (Block 208, Cybinka – Torzym, Zielona Góra West), and in the Mesozoic of the Polish Lowlands (Koło).Keywords: concession policy, international tender, oil and gas exploration horizons, prospective areas
Procedia PDF Downloads 2122373 Median Versus Ulnar Medial Thenar Motor Recording in Diagnosis Of Carpal Tunnel Syndrome
Authors: Emmanuel Kamal Aziz Saba
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Aim of the work: This study proposed to assess the role of the median versus ulnar medial thenar motor (MTM) recording in supporting the diagnosis of carpal tunnel syndrome (CTS). Patients and methods: The present study included 130 hands (70 CTS and 60 controls). Clinical examination was done for all patients. The following tests were done (using surface electrodes recording) for patients and control: (1) sensory nerve conduction studies: median nerve, ulnar nerve and median versus ulnar digit four sensory study; (2) motor nerve conduction studies: median nerve, ulnar nerve, median (second lumbrical) versus ulnar (interosseous) (2-LINT) motor study and median versus ulnar (MTM) study. Results: The tests with higher sensitivity in diagnosing CTS were median versus ulnar (2-LINT) motor latency difference (87.1%), median versus ulnar (MTM) motor latency difference (80%) and median versus ulnar digit four sensory latency differences (91.4%). There was no statistically significant difference between median versus ulnar (MTM) motor latency difference with both median versus ulnar (2-LINT) motor latency difference and median versus ulnar digit four sensory latency difference (P > 0.05) as regards the confirmation of CTS. Conclusions: Median versus ulnar (MTM) motor latency difference has high sensitivity and specificity for the diagnosis of CTS as for both median versus ulnar (2-LINT) motor latency difference and median versus ulnar digit four sensory latency differences. It can be considered a useful neurophysiological test to be used in combination with another median versus ulnar comparative tests for confirming the diagnosis of CTS beside other well-known electrophysiological tests.Keywords: carpal tunnel syndrome, medial thenar motor, median nerve, ulnar nerve
Procedia PDF Downloads 4462372 Sensitivity to Misusing Verb Inflections in Both Finite and Non-Finite Clauses in Native and Non-Native Russian: A Self-Paced Reading Investigation
Authors: Yang Cao
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Analyzing the oral production of Chinese-speaking learners of English as a second language (L2), we can find a large variety of verb inflections – Why does it seem so hard for them to use consistent correct past morphologies in obligatory past contexts? Failed Functional Features Hypothesis (FFFH) attributes the rather non-target-like performance to the absence of [±past] feature in their L1 Chinese, arguing that for post puberty learners, new features in L2 are no more accessible. By contrast, Missing Surface Inflection Hypothesis (MSIH) tends to believe that all features are actually acquirable for late L2 learners, while due to the mapping difficulties from features to forms, it is hard for them to realize the consistent past morphologies on the surface. However, most of the studies are limited to the verb morphologies in finite clauses and few studies have ever attempted to figure out these learners’ performance in non-finite clauses. Additionally, it has been discussed that Chinese learners may be able to tell the finite/infinite distinction (i.e. the [±finite] feature might be selected in Chinese, even though the existence of [±past] is denied). Therefore, adopting a self-paced reading task (SPR), the current study aims to analyze the processing patterns of Chinese-speaking learners of L2 Russian, in order to find out if they are sensitive to misuse of tense morphologies in both finite and non-finite clauses and whether they are sensitive to the finite/infinite distinction presented in Russian. The study targets L2 Russian due to its systematic morphologies in both present and past tenses. A native Russian group, as well as a group of English-speaking learners of Russian, whose L1 has definitely selected both [±finite] and [±past] features, will also be involved. By comparing and contrasting performance of the three language groups, the study is going to further examine and discuss the two theories, FFFH and MSIH. Preliminary hypotheses are: a) Russian native speakers are expected to spend longer time reading the verb forms which violate the grammar; b) it is expected that Chinese participants are, at least, sensitive to the misuse of inflected verbs in non-finite clauses, although no sensitivity to the misuse of infinitives in finite clauses might be found. Therefore, an interaction of finite and grammaticality is expected to be found, which indicate that these learners are able to tell the finite/infinite distinction; and c) having selected [±finite] and [±past], English-speaking learners of Russian are expected to behave target-likely, supporting L1 transfer.Keywords: features, finite clauses, morphosyntax, non-finite clauses, past morphologies, present morphologies, Second Language Acquisition, self-paced reading task, verb inflections
Procedia PDF Downloads 1102371 IL6/PI3K/mTOR/GFAP Molecular Pathway Role in COVID-19-Induced Neurodegenerative Autophagy, Impacts and Relatives
Authors: Mohammadjavad Sotoudeheian
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COVID-19, which began in December 2019, uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter and spread through the cells. ACE2 mRNA is present in almost every organ, including nasopharynx, lung, as well as the brain. Ports of entry of SARS-CoV-2 into the central nervous system (CNS) may include arterial circulation, while viremia is remarkable. However, it is imperious to develop neurological symptoms evaluation CSF analysis in patients with COVID-19, but theoretically, ACE2 receptors are expressed in cerebellar cells and may be a target for SARS-CoV-2 infection in the brain. Recent evidence agrees that SARS-CoV-2 can impact the brain through direct and indirect injury. Two biomarkers for CNS injury, glial fibrillary acidic protein (GFAP) and neurofilament light chain (NFL) detected in the plasma of patients with COVID-19. NFL, an axonal protein expressed in neurons, is related to axonal neurodegeneration, and GFAP is over-expressed in CNS inflammation. GFAP cytoplasmic accumulation causes Schwan cells to misfunction, so affects myelin generation, reduces neuroskeletal support over NfLs during CNS inflammation, and leads to axonal degeneration. Interleukin-6 (IL-6), which extensively over-express due to interleukin storm during COVID-19 inflammation, regulates gene expression, as well as GFAP through STAT molecular pathway. IL-6 also impresses the phosphoinositide 3-kinase (PI3K)/STAT/smads pathway. The PI3K/ protein kinase B (Akt) pathway is the main modulator upstream of the mammalian target of rapamycin (mTOR), and alterations in this pathway are common in neurodegenerative diseases. Most neurodegenerative diseases show a disruption of autophagic function and display an abnormal increase in protein aggregation that promotes cellular death. Therefore, induction of autophagy has been recommended as a rational approach to help neurons clear abnormal protein aggregates and survive. The mTOR is a major regulator of the autophagic process and is regulated by cellular stressors. The mTORC1 pathway and mTORC2, as complementary and important elements in mTORC1 signaling, have become relevant in the regulation of the autophagic process and cellular survival through the extracellular signal-regulated kinase (ERK) pathway.Keywords: mTORC1, COVID-19, PI3K, autophagy, neurodegeneration
Procedia PDF Downloads 862370 PYTHEIA: A Scale for Assessing Rehabilitation and Assistive Robotics
Authors: Yiannis Koumpouros, Effie Papageorgiou, Alexandra Karavasili, Foteini Koureta
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The objective of the present study was to develop a scale called PYTHEIA. The PYTHEIA is a self-reported measure for the assessment of rehabilitation and assistive robotics and other assistive technology devices. The development of PYTHEIA faced the absence of a valid instrument that can be used to evaluate the assistive robotic devices both as a whole, as well as any of their individual components or functionalities implemented. According to the results presented, PYTHEIA is a valid and reliable scale able to be applied to different target groups for the subjective evaluation of various assistive technology devices.Keywords: rehabilitation, assistive technology, assistive robots, rehabilitation robots, scale, psychometric test, assessment, validation, user satisfaction
Procedia PDF Downloads 3122369 Sheep Pox Virus Recombinant Proteins To Develop Subunit Vaccines
Authors: Olga V. Chervyakova, Elmira T. Tailakova, Vitaliy M. Strochkov, Kulyaisan T. Sultankulova, Nurlan T. Sandybayev, Lev G. Nemchinov, Rosemarie W. Hammond
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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 2422368 Enzyme Immobilization: A Strategy to Overcome Enzyme Limitations and Expand Their Applications
Authors: Charline Monnier, Rudolf Andrys, Irene Castellino, Lucie Zemanova
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Due to their inherent sustainability and compatibility with green chemistry principles, enzymes are attracting increasing attention for various applications like bioremediation or biocatalysis. These natural catalysts boast remarkable substrate specificity and operate under mild biological conditions. However, their intrinsic limitations, such as instability at high temperatures or in organic solvents, impede their wider applicability. Enzyme immobilization on supportive matrices emerges as a promising strategy to address these challenges. This approach not only facilitates enzyme reusability but also offers the potential to modulate their stability, activity, and selectivity. The present study investigates the immobilization and application of two distinct groups of hydrolases on supportive matrices: PETases, naturally capable of PolyEthylene Terephthalate (PET) degradation, and cholinesterases (ChEs), key enzymes in neurotransmitter regulation. All tested enzymes will be immobilized on porous and non-porous particles using both covalent and non-covalent methods. Additionally, the stability of PETases and cholinesterases will be explored, followed by exposure to denaturing conditions to assess their resilience under harsh conditions. Furthermore, due to the exceptional catalytic efficiency and selectivity, their biocatalytic efficiency will be tested using xenobiotic substrates, aiming to establish them as replacements for conventional chemical catalysts in environmentally friendly processes. By exploiting the power of enzyme immobilization, this research strives to unlock the full potential of these biocatalysts for sustainable and efficient technological advancements.Keywords: biocatalysis, bioremediation, enzyme efficiency, enzyme immobilization, green chemistry
Procedia PDF Downloads 582367 Improving the Bioprocess Phenotype of Chinese Hamster Ovary Cells Using CRISPR/Cas9 and Sponge Decoy Mediated MiRNA Knockdowns
Authors: Kevin Kellner, Nga Lao, Orla Coleman, Paula Meleady, Niall Barron
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Chinese Hamster Ovary (CHO) cells are the prominent cell line used in biopharmaceutical production. To improve yields and find beneficial bioprocess phenotypes genetic engineering plays an essential role in recent research. The miR-23 cluster, specifically miR-24 and miR-27, was first identified as differentially expressed during hypothermic conditions suggesting a role in proliferation and productivity in CHO cells. In this study, we used sponge decoy technology to stably deplete the miRNA expression of the cluster. Furthermore, we implemented the CRISPR/Cas9 system to knockdown miRNA expression. Sponge constructs were designed for an imperfect binding of the miRNA target, protecting from RISC mediated cleavage. GuideRNAs for the CRISPR/Cas9 system were designed to target the seed region of the miRNA. The expression of mature miRNA and precursor were confirmed using RT-qPCR. For both approaches stable expressing mixed populations were generated and characterised in batch cultures. It was shown, that CRISPR/Cas9 can be implemented in CHO cells with achieving high knockdown efficacy of every single member of the cluster. Targeting of one miRNA member showed that its genomic paralog is successfully targeted as well. The stable depletion of miR-24 using CRISPR/Cas9 showed increased growth and specific productivity in a CHO-K1 mAb expressing cell line. This phenotype was further characterized using quantitative label-free LC-MS/MS showing 186 proteins differently expressed with 19 involved in proliferation and 26 involved in protein folding/translation. Targeting miR-27 in the same cell line showed increased viability in late stages of the culture compared to the control. To evaluate the phenotype in an industry relevant cell line; the miR-23 cluster, miR-24 and miR-27 were stably depleted in a Fc fusion CHO-S cell line which showed increased batch titers up to 1.5-fold. In this work, we highlighted that the stable depletion of the miR-23 cluster and its members can improve the bioprocess phenotype concerning growth and productivity in two different cell lines. Furthermore, we showed that using CRISPR/Cas9 is comparable to the traditional sponge decoy technology.Keywords: Chinese Hamster ovary cells, CRISPR/Cas9, microRNAs, sponge decoy technology
Procedia PDF Downloads 2002366 Exploring the Effects of Transcendental Mindfulness Meditation on Anxiety Symptoms in Young Females
Authors: Claudia Cedeno Nadal, Mei-Ling Villafana, Griela Rodriguez, Jessica Martin, Jennifer Martin, Megan Patel
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This study systematically examines the impact of Transcendental Mindfulness Meditation on anxiety symptoms in young females aged 18-25. Through a comprehensive literature review, we found consistent evidence supporting the positive influence of Transcendental Mindfulness Meditation on reducing anxiety, enhancing overall well-being, and decreasing perceived stress levels within this demographic. The mechanisms underlying these effects include heightened self-awareness, improved emotional regulation, and the development of effective stress-coping strategies. These findings have significant implications for mental health interventions targeting young females. However, the reviewed studies had some limitations, such as small sample sizes and reliance on self-report measures. To advance this field, future research should focus on larger sample sizes and utilize a broader range of measurement methods, including neuroscience assessments. Additionally, investigating the temporal relationships between Transcendental Mindfulness Meditation, proposed mediators, and anxiety symptoms will help establish causal specificity and a deeper understanding of the precise mechanisms of action. The development of integrative models based on these mechanisms can further enhance the effectiveness of Transcendental Mindfulness Meditation as an intervention for anxiety in this demographic. This study contributes to the current knowledge on the potential benefits of Transcendental Mindfulness Meditation for reducing anxiety in young females, paving the way for more targeted and effective mental health interventions in this population.Keywords: mindfulness, meditation, anxiety, transcendental mindfulness
Procedia PDF Downloads 712365 The Acquisition of /r/ By Setswana-Learning Children
Authors: Keneilwe Matlhaku
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Crosslinguistic studies (theoretical and clinical) have shown delays and significant misarticulation in the acquisition of the rhotics. This article provides a detailed analysis of the early development of the rhotic phoneme, an apical trill /r/, by monolingual Setswana (Tswana S30) children of age ranges between 1 and 4 years. The data display the following trends: (1) late acquisition of /r/; (2) a wide range of substitution patterns involving this phoneme (i.e., gliding, coronal stopping, affrication, deletion, lateralization, as well as, substitution to a dental and uvular fricative). The primary focus of the article is on the potential origins of these variations of /r/, even within the same language. Our data comprises naturalistic longitudinal audio recordings of 6 children (2 males and 4 females) whose speech was recorded in their homes over a period of 4 months with no or only minimal disruptions in their daily environments. Phon software (Rose et al. 2013; Rose & MacWhinney 2014) was used to carry out the orthographic and phonetic transcriptions of the children’s data. Phon also enabled the generation of the children’s phonological inventories for comparison with adult target IPA forms. We explain the children’s patterns through current models of phonological emergence (MacWhinney 2015) as well as McAllister Byun, Inkelas & Rose (2016); Rose et al., (2022), which highlight the perceptual and articulatory factors influencing the development of sounds and sound classes. We highlight how the substitution patterns observed in the data can be captured through a consideration of the auditory properties of the target speech sounds, combined with an understanding of the types of articulatory gestures involved in the production of these sounds. These considerations, in turn, highlight some of the most central aspects of the challenges faced by the child toward learning these auditory-articulatory mappings. We provide a cross-linguistic survey of the acquisition of rhotic consonants in a sample of related and unrelated languages in which we show that the variability and volatility in the substitution patterns of /r/ is also brought about by the properties of the children’s ambient languages. Beyond theoretical issues, this article sets an initial foundation for developing speech-language pathology materials and services for Setswana learning children, an emerging area of public service in Botswana.Keywords: rhotic, apical trill, Phon, phonological emergence, auditory, articulatory, mapping
Procedia PDF Downloads 402364 A 7 Dimensional-Quantitative Structure-Activity Relationship Approach Combining Quantum Mechanics Based Grid and Solvation Models to Predict Hotspots and Kinetic Properties of Mutated Enzymes: An Enzyme Engineering Perspective
Authors: R. Pravin Kumar, L. Roopa
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Enzymes are molecular machines used in various industries such as pharmaceuticals, cosmetics, food and animal feed, paper and leather processing, biofuel, and etc. Nevertheless, this has been possible only by the breath-taking efforts of the chemists and biologists to evolve/engineer these mysterious biomolecules to work the needful. Main agenda of this enzyme engineering project is to derive screening and selection tools to obtain focused libraries of enzyme variants with desired qualities. The methodologies for this research include the well-established directed evolution, rational redesign and relatively less established yet much faster and accurate insilico methods. This concept was initiated as a Receptor Rependent-4Dimensional Quantitative Structure Activity Relationship (RD-4D-QSAR) to predict kinetic properties of enzymes and extended here to study transaminase by a 7D QSAR approach. Induced-fit scenarios were explored using Quantum Mechanics/Molecular Mechanics (QM/MM) simulations which were then placed in a grid that stores interactions energies derived from QM parameters (QMgrid). In this study, the mutated enzymes were immersed completely inside the QMgrid and this was combined with solvation models to predict descriptors. After statistical screening of descriptors, QSAR models showed > 90% specificity and > 85% sensitivity towards the experimental activity. Mapping descriptors on the enzyme structure revealed hotspots important to enhance the enantioselectivity of the enzyme.Keywords: QMgrid, QM/MM simulations, RD-4D-QSAR, transaminase
Procedia PDF Downloads 1372363 Socializing Young Females towards Sports
Authors: Mohinder Kumar
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Sports are considered as the very prominent social institution in almost every society because it reflects the mores, values, and general culture of a society. Sports activity tend to pave the foundation for learning acceptable values and beliefs and for acquiring desirable character traits such as self-discipline, sportsmanship, and an appreciation for hard work, fairness, self-respect, leadership, followership, justice, perseverance, competition, and goal attainment. The present study focuses on ideal ways of socializing youngsters into sports. Influences of some socializing agents (e.g. family, school, community) are reviewed and suggestions made as to how these socializing agents can be oriented and made effective in carrying out functional processes toward target ends.Keywords: sports, socializing, family, community, society
Procedia PDF Downloads 4612362 Effect on Haemolymph Cellular Parameters of Periplaneta Americana Following Challenge with Agrobacterium Tumefaciens: A Possible Microbial Control Agent
Authors: Fouzia Qamar, Shahida Hasnain
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The present study is primarily concerned with the alteration in haemocyte profile of adult male Periplaneta americana with emphasis on the effect of bacterial inoculations on the haemogram i.e., total haemocyte count (THC) and differential haemocyte count (DHC) of different haemocyte types of the target insect. Haemolymph cellular profile showed considerable alterations under the effect of nine strains of Agrobacterium tumefaciens after 8, 16 and 24 hrs of treatment thereby signifying the potential role of Agrobacterium tumefaciens as a possible biocontrol agent against the house hold pests.Keywords: Agrobacterium tumefaciens, Periplaneta americana, Haemolymph, cellular parametes
Procedia PDF Downloads 3862361 Expression of miRNA 335 in Gall Bladder Cancer: A Correlative Study
Authors: Naseem Fatima, A. N. Srivastava, Tasleem Raza, Vijay Kumar
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Introduction: Carcinoma gallbladder is third most common gastrointestinal lethal disease with the highest incidence and mortality rate among women in Northern India. Scientists have found several risk factors that make a person more likely to develop gallbladder cancer; among these risk factors, deregulation of miRNAs has been demonstrated to be one of the most crucial factors. The changes in the expression of specific miRNA genes result in the control of inflammation, cell cycle regulation, stress response, proliferation, differentiation, apoptosis and invasion thus mediate the process in tumorgenesis. The aim of this study was to investigate the role of MiRNA-335 and may as a molecular marker in early detection of gallbladder cancer in suspected cases. Material and Methods: A total of 20 consecutive patients with gallbladder cancer aged between 30-75 years were registered for the study. Total RNA was extracted from tissue by using the mirVANA MiRNA isolation Kit according to the manufacturer’s protocol. The MiRNA- 335 and U6 snRNA-specific cDNA were reverse-transcribed from total RNA using Taqman microRNA reverse-transcription kit according to the manufacturer’s protocol. TaqMan MiRNA probes hsa-miR-335 and Taqman Master Mix without AmpEase UNG, Individual real-time PCR assays were performed in a 20 μL reaction volume on a Real-Time PCR system (Applied Biosystems StepOnePlus™) to detect MiRNA-335 expression in tissue. Relative quantification of target MiRNA expression was evaluated using the comparative cycle threshold (CT) method. The correlation was done in between cycle threshold (CT Value) of target MiRNA in gallbladder cancer with respect to non-cancerous Cholelithiasis gallbladder. Each sample was examined in triplicate. The Newman-Keuls Multiple Comparison Test was used to determine the expression of miR-335. Results: MiRNA335 was found to be significantly downregulated in the gallbladder cancer tissue (P<0.001), when compared with non-cancerous Cholelithiasis gallbladder cases. Out of 20 cases, 75% showed reduced expression of MiRNA335, were at last stage of disease with low overall survival rate and remaining 25% were showed up-regulated expression of MiRNA335 with high survival rate. Conclusion: The present study showed that reduced expression of MiRNA335 is associated with the advancement of the disease, and its deregulation may provide important clues to understanding it as a prognostic marker and opportunities for future research.Keywords: carcinoma gallbladder, downregulation, MiRNA-335, RT-PCR assay
Procedia PDF Downloads 3612360 Use of a Chagas Urine Nanoparticle Test (Chunap) to Correlate with Parasitemia Levels in T. cruzi/HIV Co-Infected Patients
Authors: Yagahira E. Castro-Sesquen, Robert H. Gilman, Carolina Mejia, Daniel E. Clark, Jeong Choi, Melissa J. Reimer-Mcatee, Rocio Castro, Jorge Flores, Edward Valencia-Ayala, Faustino Torrico, Ricardo Castillo-Neyra, Lance Liotta, Caryn Bern, Alessandra Luchini
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Early diagnosis of reactivation of Chagas disease in HIV patients could be lifesaving; however, in Latin American the diagnosis is performed by detection of parasitemia by microscopy which lacks sensitivity. To evaluate if levels of T. cruzi antigens in urine determined by Chunap (Chagas urine nanoparticle test) are correlated with parasitemia levels in T. cruzi/HIV co-infected patients. T. cruzi antigens in urine of HIV patients (N=55: 31 T. cruzi infected and 24 T. cruzi serology negative) were concentrated using hydrogel particles and quantified by Western Blot and a calibration curve. The percentage of Chagas positive patients determined by Chunap compared to blood microscopy, qPCR, and ELISA was 100% (6/6), 95% (18/19) and 74% (23/31), respectively. Chunap specificity was 91.7%. Linear regression analysis demonstrated a direct relationship between parasitemia levels (determined by qPCR) and urine T. cruzi antigen concentrations (p<0.001). A cut-off of > 105 pg was chosen to determine patients with reactivation of Chagas disease (6/6). Urine antigen concentration was significantly higher among patients with CD4+ lymphocyte counts below 200/mL (p=0.045). Chunap shows potential for early detection of reactivation and with appropriate adaptation can be used for monitoring Chagas disease status in T. cruzi/HIV co-infected patients.Keywords: antigenuria, Chagas disease, Chunap, nanoparticles, parasitemia, poly N-isopropylacrylamide (NIPAm)/trypan blue particles (polyNIPAm/TB), reactivation of Chagas disease.
Procedia PDF Downloads 3782359 Efficacy of Agrobacterium Tumefaciens as a Possible Entomopathogenic Agent
Authors: Fouzia Qamar, Shahida Hasnain
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The objective of the present study was to evaluate the possible role of Agrobacterium tumefaciens as a possible insect biocontrol agent. Pests selected for the present challenge were adult males of Periplaneta americana and last instar larvae of Pieris brassicae and Spodoptera litura. Different ranges of bacterial doses were selected and tested to score the mortalities of the insects after 24 hours, for the lethal dose estimation studies. Mode of application for the inoculation of the bacteria, was the microinjection technique. The evaluation of the possible entomopathogenic carrying attribute of bacterial Ti plasmid, led to the conclusion that the loss of plasmid was associated with the loss of virulence against target insects.Keywords: agrobacterium tumefaciens, toxicity assessment, biopesticidal attribute, entomopathogenic agent
Procedia PDF Downloads 3802358 Comparison of Mini-BESTest versus Berg Balance Scale to Evaluate Balance Disorders in Parkinson's Disease
Authors: R. Harihara Prakash, Shweta R. Parikh, Sangna S. Sheth
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The purpose of this study was to explore the usefulness of the Mini-BESTest compared to the Berg Balance Scale in evaluating balance in people with Parkinson's Disease (PD) of varying severity. Evaluation were done to obtain (1) the distribution of patients scores to look for ceiling effects, (2) concurrent validity with severity of disease, and (3) the sensitivity & specificity of separating people with or without postural response deficits. Methods and Material: Seventy-seven(77) people with Parkinson's Disease were tested for balance deficits using the Berg Balance Scale, Mini-BESTest. Unified Parkinson’s Disease Rating Scale (UPDRS) III and the Hoehn & Yahr (H&Y) disease severity scales were used for classification. Materials used in this study were case record sheet, chair without arm rests or wheels, Incline ramp, stopwatch, a box, 3 meter distance measured out and marked on the floor with tape [from chair]. Statistical analysis used: Multiple Linear regression was carried out of UPDRS jointly on the two scores for the Berg and Mini-BESTest. Receiver operating characteristic curves for classifying people into two groups based on a threshold for the H&Y score, to discriminate between mild PD versus more severe PD.Correlation co-efficient to find relativeness between the two variables. Results: The Mini-BESTest is highly correlated with the Berg (r = 0.732,P < 0.001), but avoids the ceiling compression effect of the Berg for mild PD (skewness −0.714 Berg, −0.512 Mini-BESTest). Consequently, the Mini-BESTest is more effective than the Berg for predicting UPDRS Motor score (P < 0.001 Mini-BESTest versus P = 0.72 Berg), and for discriminating between those with and without postural response deficits as measured by the H&Y (ROC).Keywords: balance, berg balance scale, MINI BESTest, parkinson's disease
Procedia PDF Downloads 3962357 Obstetric Outcome after Hysteroscopic Septum Resection in Patients with Uterine Septa of Various Sizes
Authors: Nilanchali Singh, Alka Kriplani, Reeta Mahey, Garima Kachhawa
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Objective: Resection of larger uterine septa does improve obstetric performance but whether smaller septa need resection and their impact on obstetric outcome is not clear. We wanted to evaluate the role of septal resection of septa of various sizes in obstetric performance. Methods: This retrospective cohort study comprised of 107 patients with uterine septum. The patients were categorized on the basis of extent of uterine septum into four groups: a) Subsepta (< 1/3rd), b) Septum > 1/3 to ½, c) Septum>1/2 to whole uterine cervix, d) Septum traversing whole of uterine cavity and cervix. Out of these 107 patients, 74 could be contacted telephonically and outcomes recorded. Sensitivity and specificity of investigative modalities were calculated. Results: Infertility was seen in maximum number of cases in complete septa (100%), whereas abortions were seen more commonly, in subsepta (18%). MRI had maximum sensitivity and positive predictive value, followed by hysteron-salpingography. Tubal block, fibroid, endometriosis, pelvic adhesions, ovarian pathologies were seen in some but no definite association of these pathologies was seen with any subgroup of septa. Almost five-year follow-up was recorded in all the subgroups. Significant reduction in infertility was seen in all septal subgroup (p=0.046, 0.032 & 0.05) patients except in subsepta (< 1/3rd uterine cavity) after septum resection. Abortions were significantly reduced (p=0.048) in third subgroup (i.e. septum > ½ to upto internal os) after hysteroscopic septum resection. Take home baby rate was 33% in subsepta and around 50% in the remaining subgroups of septa. Conclusions: Septal resection improves obstetric performance in patients with uterine septa of various sizes. Whether septal resection improves obstetric performance in patients with subsepta or very small septa, is controversial. Larger studies addressing this issue need to be planned.Keywords: septal resection, obstetric outcome, infertility, septum size
Procedia PDF Downloads 3192356 Comprehensive Validation of High-Performance Liquid Chromatography-Diode Array Detection (HPLC-DAD) for Quantitative Assessment of Caffeic Acid in Phenolic Extracts from Olive Mill Wastewater
Authors: Layla El Gaini, Majdouline Belaqziz, Meriem Outaki, Mariam Minhaj
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In this study, it introduce and validate a high-performance liquid chromatography method with diode-array detection (HPLC-DAD) specifically designed for the accurate quantification of caffeic acid in phenolic extracts obtained from olive mill wastewater. The separation process of caffeic acid was effectively achieved through the use of an Acclaim Polar Advantage column (5µm, 250x4.6mm). A meticulous multi-step gradient mobile phase was employed, comprising water acidified with phosphoric acid (pH 2.3) and acetonitrile, to ensure optimal separation. The diode-array detection was adeptly conducted within the UV–VIS spectrum, spanning a range of 200–800 nm, which facilitated precise analytical results. The method underwent comprehensive validation, addressing several essential analytical parameters, including specificity, repeatability, linearity, as well as the limits of detection and quantification, alongside measurement uncertainty. The generated linear standard curves displayed high correlation coefficients, underscoring the method's efficacy and consistency. This validated approach is not only robust but also demonstrates exceptional reliability for the focused analysis of caffeic acid within the intricate matrices of wastewater, thus offering significant potential for applications in environmental and analytical chemistry.Keywords: high-performance liquid chromatography (HPLC-DAD), caffeic acid analysis, olive mill wastewater phenolics, analytical method validation
Procedia PDF Downloads 722355 Structured-Ness and Contextual Retrieval Underlie Language Comprehension
Authors: Yao-Ying Lai, Maria Pinango, Ashwini Deo
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While grammatical devices are essential to language processing, how comprehension utilizes cognitive mechanisms is less emphasized. This study addresses this issue by probing the complement coercion phenomenon: an entity-denoting complement following verbs like begin and finish receives an eventive interpretation. For example, (1) “The queen began the book” receives an agentive reading like (2) “The queen began [reading/writing/etc.…] the book.” Such sentences engender additional processing cost in real-time comprehension. The traditional account attributes this cost to an operation that coerces the entity-denoting complement to an event, assuming that these verbs require eventive complements. However, in closer examination, examples like “Chapter 1 began the book” undermine this assumption. An alternative, Structured Individual (SI) hypothesis, proposes that the complement following aspectual verbs (AspV; e.g. begin, finish) is conceptualized as a structured individual, construed as an axis along various dimensions (e.g. spatial, eventive, temporal, informational). The composition of an animate subject and an AspV such as (1) engenders an ambiguity between an agentive reading along the eventive dimension like (2), and a constitutive reading along the informational/spatial dimension like (3) “[The story of the queen] began the book,” in which the subject is interpreted as a subpart of the complement denotation. Comprehenders need to resolve the ambiguity by searching contextual information, resulting in additional cost. To evaluate the SI hypothesis, a questionnaire was employed. Method: Target AspV sentences such as “Shakespeare began the volume.” were preceded by one of the following types of context sentence: (A) Agentive-biasing, in which an event was mentioned (…writers often read…), (C) Constitutive-biasing, in which a constitutive meaning was hinted (Larry owns collections of Renaissance literature.), (N) Neutral context, which allowed both interpretations. Thirty-nine native speakers of English were asked to (i) rate each context-target sentence pair from a 1~5 scale (5=fully understandable), and (ii) choose possible interpretations for the target sentence given the context. The SI hypothesis predicts that comprehension is harder for the Neutral condition, as compared to the biasing conditions because no contextual information is provided to resolve an ambiguity. Also, comprehenders should obtain the specific interpretation corresponding to the context type. Results: (A) Agentive-biasing and (C) Constitutive-biasing were rated higher than (N) Neutral conditions (p< .001), while all conditions were within the acceptable range (> 3.5 on the 1~5 scale). This suggests that when lacking relevant contextual information, semantic ambiguity decreases comprehensibility. The interpretation task shows that the participants selected the biased agentive/constitutive reading for condition (A) and (C) respectively. For the Neutral condition, the agentive and constitutive readings were chosen equally often. Conclusion: These findings support the SI hypothesis: the meaning of AspV sentences is conceptualized as a parthood relation involving structured individuals. We argue that semantic representation makes reference to spatial structured-ness (abstracted axis). To obtain an appropriate interpretation, comprehenders utilize contextual information to enrich the conceptual representation of the sentence in question. This study connects semantic structure to human’s conceptual structure, and provides a processing model that incorporates contextual retrieval.Keywords: ambiguity resolution, contextual retrieval, spatial structured-ness, structured individual
Procedia PDF Downloads 3332354 RussiAnglicized© Slang and Translation: A Clockwork Orange Tick-Tock
Authors: Mahnaz Movahedi
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Slang argot plays a fundamental role in Burgess’ teenage special sociolect in his novel A Clockwork Orange, offered a wide variety of instances to be analyzed. Consequently, translation of the notions and keeping the effect would be of great importance. Burgess named his interesting RussiAnglicized©-slang word as Nadsat, stands for –teen, mostly derived from Russian and Cockney rhyming. The paper discusses the lexical origin and Persian translation of his weird slang words illustrating a teenage-gang argot. The product depicts creativity but mistranslation that leads to the loss of slang meaning load and atmosphere in the target text.Keywords: argot, mistranslation, slang, sociolect
Procedia PDF Downloads 2532353 The Influence of Screen Translation on Creative Audiovisual Writing: A Corpus-Based Approach
Authors: John D. Sanderson
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The popularity of American cinema worldwide has contributed to the development of sociolects related to specific film genres in other cultural contexts by means of screen translation, in many cases eluding norms of usage in the target language, a process whose result has come to be known as 'dubbese'. A consequence for the reception in countries where local audiovisual fiction consumption is far lower than American imported productions is that this linguistic construct is preferred, even though it differs from common everyday speech. The iconography of film genres such as science-fiction, western or sword-and-sandal films, for instance, generates linguistic expectations in international audiences who will accept more easily the sociolects assimilated by the continuous reception of American productions, even if the themes, locations, characters, etc., portrayed on screen may belong in origin to other cultures. And the non-normative language (e.g., calques, semantic loans) used in the preferred mode of linguistic transfer, whether it is translation for dubbing or subtitling, has diachronically evolved in many cases into a status of canonized sociolect, not only accepted but also required, by foreign audiences of American films. However, a remarkable step forward is taken when this typology of artificial linguistic constructs starts being used creatively by nationals of these target cultural contexts. In the case of Spain, the success of American sitcoms such as Friends in the 1990s led Spanish television scriptwriters to include in national productions lexical and syntactical indirect borrowings (Anglicisms not formally identifiable as such because they include elements from their own language) in order to target audiences of the former. However, this commercial strategy had already taken place decades earlier when Spain became a favored location for the shooting of foreign films in the early 1960s. The international popularity of the then newly developed sub-genre known as Spaghetti-Western encouraged Spanish investors to produce their own movies, and local scriptwriters made use of the dubbese developed nationally since the advent of sound in film instead of using normative language. As a result, direct Anglicisms, as well as lexical and syntactical borrowings made up the creative writing of these Spanish productions, which also became commercially successful. Interestingly enough, some of these films were even marketed in English-speaking countries as original westerns (some of the names of actors and directors were anglified to that purpose) dubbed into English. The analysis of these 'back translations' will also foreground some semantic distortions that arose in the process. In order to perform the research on these issues, a wide corpus of American films has been used, which chronologically range from Stagecoach (John Ford, 1939) to Django Unchained (Quentin Tarantino, 2012), together with a shorter corpus of Spanish films produced during the golden age of Spaghetti Westerns, from una tumba para el sheriff (Mario Caiano; in English lone and angry man, William Hawkins) to tu fosa será la exacta, amigo (Juan Bosch, 1972; in English my horse, my gun, your widow, John Wood). The methodology of analysis and the conclusions reached could be applied to other genres and other cultural contexts.Keywords: dubbing, film genre, screen translation, sociolect
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