Search results for: text similarity

Authors: Tapiwa Brine Mutsauri

Abstract:

The MEROPS system is an information resource for proteases that classifies them into clans according to their catalytic type. Within the Plant kingdom, cysteine ​​proteases are one of the best known, as they are the catalytic type on which the first studies on plant proteases were focused. Plant cysteine ​​proteases have a similar mechanism of action to serine proteases, and some are known to have activity on factors of the blood coagulation cascade, such as a potent antithrombotic effect, and also cause increased fibrinolysis. Of a few plant cysteine ​​proteases, the three-dimensional structure is known, so a method of interest to be able to predict their potential activity on the factors of the coagulation cascade would be to know their structure. Phylogenetics is the study of the evolutionary relationships between biological entities, often species, individuals, or genes (which can be called taxa). It is essential to identify the evolutionary position and the possible distribution of these enzymes in the plant kingdom, particularly those that act on coagulation factors. Bioinformatic tools, such as Clustal Omega / Jalview and Mega6, can be used to create phylogenetic trees. From the results of the alignment, it can be seen that although there is a certain degree of conservation (Conservation) and consensus (Consensus) among the eleven sequences, the functionally important motifs (those corresponding to the active site), the degree of conservation and consensus is very low. We could then infer that although activity on coagulation is reported for these enzymes, linked to their structural and mechanistic similarity with serine proteases, this activity may not have a direct or primary relationship with the proteolytic activity associated with their common, poorly conserved active site in this case. This ultimately could be related to modifications in the reaction mechanism of several of the enzymes studied, which would require more detailed study. Also, below, we will deal with factors that may have a greater influence on this result. The results of this work enrich the understanding of how species (and molecular sequences in general) evolve. Through phylogenetics, we learn not only how sequences came to be the way they are today but also the general principles that allow us to predict how they will change in the future. For pharmaceutical sciences, phylogenetic selection of biologically related species can help identify closely related members of a species with compounds of pharmacological importance, such as plant cysteine ​​proteases, in addition to identifying structural features that may influence their pharmacological activity and which can be valuable for drug design.

Keywords: computational simulation, proteases, coagulation, bioinformatics

Procedia PDF Downloads 16

Authors: Zdzislaw Kaminski, Zbigniew Czyz

Abstract:

The paper discusses the results of the research into a wind turbine with a vertical axis of rotation which was performed with the open return wind tunnel, Gunt HM 170, at the laboratory of the Department of Thermodynamics, Fluid Mechanics and Propulsion Aviation Systems of Lublin University of Technology. Wind tunnel experiments are a necessary step to construct any new type of wind turbine, to validate design assumptions and numerical results. This research focused on the rotor with the blades capable of modifying their working surfaces, i.e. absorbing wind kinetic energy. The operation of this rotor is based on adjusting angular aperture α of the top and bottom parts of the blades mounted on an axis. If this angle α increases, the working surface which absorbs wind kinetic energy also increases. The study was performed on scaled and geometrically similar models with the criteria of similarity relevant for the type of research preserved. The rotors with varied angular apertures of their blades were printed for the research with a powder 3D printer, ZPrinter® 450. This paper presents the research results for the selected flow speed of 6.5 m/s for the three angular apertures of the rotor blades, i.e. 30°, 60°, 90° at varied speeds. The test stand enables the turbine rotor to be braked to achieve the required speed and airflow speed and torque to be recorded. Accordingly, the torque and power as a function of airflow were plotted. The rotor with its adjustable blades enables turbine power to be adjusted within a wide range of wind speeds. A variable angular aperture of blade working surfaces α in a wind turbine enables us to control the speed of the turbine and consequently its output power. Reducing the angular aperture of working surfaces results in reduced speed, and if a special current generator applied, electrical output power is reduced, too. Speed adjusted by changing angle α enables the maximum load acting on rotor blades to be controlled. The solution under study is a kind of safety against a damage of a turbine due to possible high wind speed.

Keywords: drive torque, renewable energy, power, wind turbine, wind tunnel

Procedia PDF Downloads 258

Authors: Patarasuda Chaisupa, R. Clay Wright

Abstract:

The selection of appropriate biomolecular targets is a crucial aspect of biopharmaceutical development. The Auxin-Inducible Degron Degradation (AID) technology has demonstrated remarkable potential in efficiently and rapidly degrading target proteins, thereby enabling the identification and acquisition of drug targets. The AID system also offers a viable method to deplete specific proteins, particularly in cases where the degradation pathway has not been exploited or when the adaptation of proteins, including the cell environment, occurs to compensate for the mutation or gene knockout. In this study, we have engineered an improved AID system tailored to deplete proteins of interest. This AID construct combines the auxin-responsive E3 ubiquitin ligase binding domain, AFB2, and the substrate degron, IAA17, fused to the target genes. Essential genes of fungi with the lowest percent amino acid similarity to human and plant orthologs, according to the Basic Local Alignment Search Tool (BLAST), were cloned into the AID construct in S. cerevisiae (AID-tagged strains) using a modular yeast cloning toolkit for multipart assembly and direct genetic modification. Each E3 ubiquitin ligase and IAA17 degron was fused to a fluorescence protein, allowing for real-time monitoring of protein levels in response to different auxin doses via cytometry. Our AID system exhibited high sensitivity, with an EC50 value of 0.040 µM (SE = 0.016) for AFB2, enabling the specific promotion of IAA17::target protein degradation. Furthermore, we demonstrate how this improved AID system enhances quantitative functional studies of various proteins in fungi. The advancements made in auxin-inducible protein degradation in this study offer a powerful approach to investigating critical target protein viability in fungi, screening protein targets for drugs, and regulating intracellular protein abundance, thus revolutionizing the study of protein function underlying a diverse range of biological processes.

Keywords: synthetic biology, bioengineering, molecular biology, biotechnology

Procedia PDF Downloads 92

Authors: Narayan Moger, Divya B., Preethi Jambagi, Krishnaveni C. K., Apsana M. R., B. R. Patil, Basvaraj Bagewadi

Abstract:

Salinity is one of the major threats to world food security. Considering the requirement for salt tolerant crop plants in the present study was undertaken to clone and transferred the salt tolerant ectA gene from marine ecosystem into agriculture crop system to impart salinity tolerance. Ectoine is the compatible solute which accumulates in the cell membrane, is known to be involved in salt tolerance activity in most of the Halophiles. The present situation is insisting to development of salt tolerant transgenic lines to combat abiotic stress. In this background, the investigation was conducted to develop transgenic tomato lines by cloning and transferring of ectA gene is an ectoine derivative capable of enzymatic action for the production of acetyl-diaminobutyric acid. The gene ectA is involved in maintaining the osmotic balance of plants. The PCR amplified ectA gene (579bp) was cloned into T/A cloning vector (pTZ57R/T). The construct pDBJ26 containing ectA gene was sequenced by using gene specific forward and reverse primers. Sequence was analyzed using BLAST algorithm to check similarity of ectA gene with other isolates. Highest homology of 99.66 per cent was found with ectA gene sequences of isolates Halomonas elongata with the available sequence information in NCBI database. The ectA gene was further sub cloned into pRI101-AN plant expression vector and transferred into E. coli DH5α for its maintenance. Further pDNM27 was mobilized into A. tumefaciens LBA4404 through tri-parental mating system. The recombinant Agrobacterium containing pDNM27 was transferred into tomato plants through In planta plant transformation method. Out of 300 seedlings, co-cultivated only twenty-seven plants were able to well establish under the greenhouse condition. Among twenty-seven transformants only twelve plants showed amplification with gene specific primers. Further work must be extended to evaluate the transformants at T1 and T2 generations for ectoine accumulation, salinity tolerance, plant growth and development and yield.

Keywords: salinity, computable solutes, ectA, transgenic, in planta transformation

Procedia PDF Downloads 81

Authors: Arpit Kumar Shrivastava, Subrat Kumar, Rajani Kanta Mohapatra, Priyadarshi Soumyaranjan Sahu

Abstract:

Computational approaches to predict structure, function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are not effective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical epitopic protein in C. hominis genome through BLASTP analysis. A 3D model of the hypothetical protein was generated using I-Tasser server through threading methodology. The quality of the model was validated through Ramachandran plot by PROCHECK server. The functional annotation of the hypothetical protein through DALI server revealed structural similarity with human Transportin 3. Phylogenetic analysis for this hypothetical protein also showed C. hominis hypothetical protein (CUV04613) was the closely related to human transportin 3 protein. The 3D protein model is further subjected to virtual screening study with inhibitors from the Zinc Database by using Dock Blaster software. Docking study reported N-(3-chlorobenzyl) ethane-1,2-diamine as the best inhibitor in terms of docking score. Docking analysis elucidated that Leu 525, Ile 526, Glu 528, Glu 529 are critical residues for ligand–receptor interactions. The molecular dynamic simulation was done to access the reliability of the binding pose of inhibitor and protein complex using GROMACS software at 10ns time point. Trajectories were analyzed at each 2.5 ns time interval, among which, H-bond with LEU-525 and GLY- 530 are significantly present in MD trajectories. Furthermore, antigenic determinants of the protein were determined with the help of DNA Star software. Our study findings showed a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for control as well as prevention of cryptosporidiosis among humans and animals.

Keywords: cryptosporidium hominis, hypothetical protein, molecular docking, molecular dynamics simulation

Procedia PDF Downloads 365

Authors: O. Yeret

Abstract:

Visual resources are all around us, especially in today's media-driven world, which gravitates, more and more, towards the visual. As a result, authentic resources, such as television advertisements, become testaments – authentic cultural materials – that reflect the landscape of certain groups and communities during a specific point in time. Engaging language students with popular advertisements can provide a great opportunity for developing cultural awareness, a component that is sometimes overlooked in the foreign language classroom. This paper will showcase practical examples of using Israeli Television Ads in various Modern Hebrew language courses. Several approaches for combining the study of language and culture, through the use of advertisements, will be included; for example, targeted assignments based on students' proficiency levels, such as: asking to recognize vocabulary words and answer basic information questions, as opposed to commenting on the significance of an ad and analyzing its particular cultural elements. The use of visual resources in the language classroom does not only enable students to learn more about the culture of the target language, but also to combine their language skills. Most often, interacting with an ad requires close listening and some reading (through captions or other data). As students analyze the ad, they employ their writing and speaking skills by answering questions in text or audio form. Hence, these interactions are able to elicit complex language use across the four domains: listening, speaking, writing, and reading. This paper will include examples of practical assignments that were developed for several Modern Hebrew language courses, together with the specific advertisements and questions related to them. Conclusions from the process and recent feedback notes received from students regarding the use of visual resources will be mentioned as well.

Keywords: authentic materials, cultural awareness, second language acquisition, visual resources

Procedia PDF Downloads 108

Authors: Ahmedou Moulaye Idriss, Tfeil Yahya, Tamas Ungi, Gabor Fichtinger

Abstract:

Image-guided therapy (IGT) plays a crucial role in minimally invasive procedures for liver interventions. Accurate segmentation of the liver and precise needle placement is essential for successful interventions such as needle aspiration. In this study, we propose a deep learning-based liver 3D slicer designed to enhance segmentation accuracy and facilitate needle aspiration procedures. The developed 3D slicer leverages state-of-the-art convolutional neural networks (CNNs) for automatic liver segmentation in medical images. The CNN model is trained on a diverse dataset of liver images obtained from various imaging modalities, including computed tomography (CT) and magnetic resonance imaging (MRI). The trained model demonstrates robust performance in accurately delineating liver boundaries, even in cases with anatomical variations and pathological conditions. Furthermore, the 3D slicer integrates advanced image registration techniques to ensure accurate alignment of preoperative images with real-time interventional imaging. This alignment enhances the precision of needle placement during aspiration procedures, minimizing the risk of complications and improving overall intervention outcomes. To validate the efficacy of the proposed deep learning-based 3D slicer, a comprehensive evaluation is conducted using a dataset of clinical cases. Quantitative metrics, including the Dice similarity coefficient and Hausdorff distance, are employed to assess the accuracy of liver segmentation. Additionally, the performance of the 3D slicer in guiding needle aspiration procedures is evaluated through simulated and clinical interventions. Preliminary results demonstrate the effectiveness of the developed 3D slicer in achieving accurate liver segmentation and guiding needle aspiration procedures with high precision. The integration of deep learning techniques into the IGT workflow shows great promise for enhancing the efficiency and safety of liver interventions, ultimately contributing to improved patient outcomes.

Keywords: deep learning, liver segmentation, 3D slicer, image guided therapy, needle aspiration

Procedia PDF Downloads 48

Authors: Song Hung Chi, Lee Chun Benn

Abstract:

Follow by the “A Research on Types of Settlement Housing Alterations of Chinese New Village in Malaysia- A Study in Ampang New Village, Selangor” preliminary informed that the main factors for expansion and enlargement suitably due to the needs of user's life and restoration purpose. The alterations behavior generally derived at the rear position of main house with different types of derivatives, the averages expansion area are not exceeding of 100㎡, while building materials used were wooden, wooden structure, and zinc which are non-permanent building materials. Therefore, a subsequent studies taken in this paper, further to analyze the drawing with summarize method, to explore the derived forms and the constituted rules of housing alterations in Ampang Village, as a more complete presentation of housing alterations in New Village. Firstly, classified the existing housing alterations into three types by using summarize method, which are Type 1, Additional of Prototype House; Type 2, Expansion of Prototype House; and Type 3, Diffusion of Additional. The results shows that the derivative mode of alterations can be divided into the use of "continuous wall" or "non-continuous wall," this will affects the structural systems and roof styles of alterations, and formed the different layers of interior space with "stages" and "continuity". On the aspects of spatial distribution, sacrificial area as a prescriptive function of space, it was mostly remains in the original location which in the center of living area after alterations. It is an important characteristic in a New Village house, reflecting the traditional Ethics of Hakka Chinese communities in the settlement. In addition, wooden as the main building materials of constituted rules for the prototype house, although there were appeared other building materials, such as cement, brick, glass, metal and zinc after alterations, but still mostly as "wooden house" pattern. Result show because of the economy of village does not significantly improve, and also forming the similarity types in alterations and constructions of the additional building with the existing. It did not significantly improve on the quality of living, but only increased the area of usage space.

Keywords: Ampang new village, derived forms, constituted rules, alterations

Procedia PDF Downloads 320

Authors: Nor Hafizah Anuar, M. Gul Akdeniz

Abstract:

This paper attempted on emphasize on the station buildings façade elements. Station buildings were essential part of the transportation that reflected the technology. Comparative analysis on architectural styles will also be made between the railway station buildings of Malaysia and any railway station buildings which have similarities. The Malay Peninsula which is strategically situated between the Straits of Malacca and the South China Sea makes it an ideal location for trade. Malacca became an important trading port whereby merchants from around the world stopover to exchange various products. The Portuguese ruled Malacca for 130 years (1511–1641) and for the next century and a half (1641–1824), the Dutch endeavoured to maintain an economic monopoly along the coasts of Malaya. Malacca came permanently under British rule under the Anglo-Dutch Treaty, 1824. Up to Malaysian independence in 1957, Malaya saw a great influx of Chinese and Indian migrants as workers to support its growing industrial needs facilitated by the British. The growing tin ore mining and rubber industry resulted as the reason of the development of the railways as urgency to transport it from one place to another. The existence of railway transportation becomes more significant when the city started to bloom and the British started to build grandeur buildings that have different functions; administrative buildings, town and city halls, railway stations, public works department, courts, and post offices.

Keywords: Malaysia, station building, architectural styles, facade elements

Procedia PDF Downloads 166

Authors: Lai Wenfang

Abstract:

Tthis study will describe how to use artificial intelligence (AI) technology to build the user-oriented platform for integrated archival service. The platform will be launched in 2020 by the National Archives Administration (NAA) in Taiwan. With the progression of information communication technology (ICT) the NAA has built many systems to provide archival service. In order to cope with new challenges, such as new ICT, artificial intelligence or blockchain etc. the NAA will try to use the natural language processing (NLP) and machine learning (ML) skill to build a training model and propose suggestions based on the data sent to the platform. NAA expects the platform not only can automatically inform the sending agencies’ staffs which records catalogues are against the transfer or destroy rules, but also can use the model to find the details hidden in the catalogues and suggest NAA’s staff whether the records should be or not to be, to shorten the auditing time. The platform keeps all the users’ browse trails; so that the platform can predict what kinds of archives user could be interested and recommend the search terms by visualization, moreover, inform them the new coming archives. In addition, according to the Archives Act, the NAA’s staff must spend a lot of time to mark or remove the personal data, classified data, etc. before archives provided. To upgrade the archives access service process, the platform will use some text recognition pattern to black out automatically, the staff only need to adjust the error and upload the correct one, when the platform has learned the accuracy will be getting higher. In short, the purpose of the platform is to deduct the government digital transformation and implement the vision of a service-oriented smart government.

Keywords: artificial intelligence, natural language processing, machine learning, visualization

Procedia PDF Downloads 174

Authors: Jakub Młoźniak, Adam Szymański, Gabriela Stondzik, Dagny Krankowska, Tomasz Mikuła

Abstract:

Nontuberculous mycobacteria (NTM) are bacterial species that cause diversely manifesting diseases mainly in immunocompromised patients. In people with HIV, NTM infection is an AIDS-defining disease and usually appears when the lymphocyte T CD4 count is below 50 cells/μl. The usage of antiretroviral therapy has decreased the prevalence of NTM among people with HIV, but the disease can still be observed especially among patients with late HIV diagnosis. Common presence in environment, human colonization, clinical similarity with tuberculosis and slow growth on culture makes NTM especially hard to diagnose. The study aimed to analyze the epidemiology and clinical course of NTM among patients with HIV. This study included patients with NTM and HIV admitted to our department between 2017 and 2023. Medical records of patients were analyzed and data on age, sex, median time from HIV diagnosis to identification of NTM infection, median CD4 count at NTM diagnosis, methods of determining NTM infection, type of species of mycobacteria identified, clinical symptoms and treatment course were gathered. Twenty-four patients (20 men, 4 women) with identified NTM were included in this study. Among them, 20 were HIV late presenters. The patients' median age was 40. The main symptoms which patients presented were fever, weight loss and cough. Pulmonary disease confirmed with positive cultures from sputum/bronchoalveolar lavage was present in 18 patients. M. avium was the most common species identified. M. marinum caused disseminated skin lesions in 1 patient. Out of all, 5 people were not treated for NTM caused by lack of symptoms and suspicion of colonization with mycobacterium. Concomitant tuberculosis was present in 6 patients. The median diagnostic time from HIV to NTM infections was 3.5 months. The median CD4 count at NTM identification was 69.5 cells/μl. Median NTM treatment time was 16 months but 7 patients haven’t finished their treatment yet. The most commonly used medications were ethambutol and clarithromycin. Among analyzed patients, 4 of them have died. NTM infections are still an important disease among patients who are HIV late presenters. This disease should be taken into consideration during the differential diagnosis of fever, weight loss and cough in people with HIV with lymphocyte T CD4 count <100 cells/μl. Presence of tuberculosis does not exclude nontuberculous mycobacterium coinfection.

Keywords: mycobacteriosis, HIV, late presenter, epidemiology

Procedia PDF Downloads 41

Authors: Hiripurage Mallika Sandamali Dissanayaka

Abstract:

Butterflies are found everywhere on Earth, helping flowering plants reproduce through pollination. Wetlands perform many valuable functions such as providing wildlife habitat. Diyasaru Wetland Park was chosen as the study site. It is located in a highly urbanized area of Sri Jayawardenepura Kotte, Sri Lanka. A distribution map was prepared to increase butterfly habitat in the urbanized area, and research was conducted to determine the most suitable sections for using it. As this wetland has footpaths for walking, line transect surveys were used to mark species within the sampling area, and directly observed species were recorded. All data collection was done from 0900 to 1200 hours and 1300 to 1600 hours and fieldwork was done from 11 February 2020 to 20 January 2021. ED binoculars (10.5x45), DSLR cameras (Canon EOS/EFS5 mm 3.5-5.6), and Garmin GPS (Etrex 10) were used to observe butterfly species, identify locations, and take photographs as evidence. Analyzing their habitats using GIS (ArcGIS Pro) to identify their distribution within the park premises, the distribution density of the known size of the population was calculated for each point by kernel density, and local similarity values were calculated for each pair of corresponding features through hotspot analysis, and cell values were determined by inverse distance weighting (IDW) using a linearly weighted combination of a set of sample points. According to the maps prepared to predict the distribution of butterflies in this park, the high level of distribution or favorable areas were near flower gardens and meadows, but some individual species prefer habitats that are more suitable for their life activities, so they live in other areas. Sixty-six (66) species belonging to six (6) families have been recorded in the premises. Sixty (60) species of least concern (LC), two (2) near threatened (NT), and four (4) vulnerable (VU) species have been recorded, and several new species, such as Plum Judy (Abisara echerius), were reported. The outcome of the study will form the basis for decision-making by the Sri Lanka Land Development (SLLD) Corporation for the future development and maintenance of the park.

Keywords: wetland, Lepidoptera, habitat, urban, west

Procedia PDF Downloads 49

Authors: Tatsiana Haiden

Abstract:

The role of the translator in the publishing process has been underestimated for centuries. Any translation is produced in a certain socio-political context by agents with different background, interests, and opinions, i.e., no translation is neutral. Any translation goes beyond the text; it is not only an interlingual transfer of signs but a social phenomenon. The case study shows how Jewish social networks influence publishing translations and aims to explain the unexpected success of the Jewish publishing house in pre-Anschluss Austria. The research shows that translators play a central role (‘Translator’s visibility’ - Pym, ‘Activist turn’ - Wolf, ‘Translator studies’ - Chesterman) in choosing what has to be translated and establishing communication between the author and the publisher. The concept of Translationskultur of Prunc is being historized and applied to the publishing house for the first time by analyzing business correspondence between the main actors of translation (publisher-translator-author). The translation studies project has become interdisciplinary –it encompasses sociology (concepts of Bourdieu’s ‘Field theory’ are used) and history. The historical research method Histoire croiseé is being used to avoid subjectivity and to introduce a new ‘translator-oriented’ vision in translation studies instead of the author-oriented one. In the course of the archival research, it was established that Jewish background plays an essential role in the destiny of the translators and the publishing house, so the Jewish studies have been added to the project. The study goes beyond the Austrian translational culture; it can be used as an example of dealing with publishing houses policies, publishing translations, and translator studies.

Keywords: history of translation, Jewish studies, publishing translations, translation sociology, translator studies, translators as actors

Procedia PDF Downloads 157

Authors: Jane Larkin

Abstract:

Combining elements of philosophy, science, and creative writing, this investigation explores how a philosophically structured science-fiction novel can challenge the theory of linearity and singularity of time through the ‘many worlds’ theory. This concept is addressed through the creation of a research exegesis and accompanying creative artefact, designed to be read in conjunction with each other in an explorative, interwoven manner. Research undertaken into scientific concepts, such as the ‘many worlds’ interpretation of quantum mechanics and diverse philosophers and their ideologies on time, is embodied in an original science-fiction narrative titled, It Goes On. The five frames that make up the creative artefact are enhanced not only by five leading philosophers and their philosophies on time but by an appreciation of the research, which comes first in the paper. Research into traditional approaches to storytelling is creatively and innovatively inverted in several ways, thus challenging the singularity and linearity of time. Further nonconventional approaches to literary techniques include an abstract narrator, embodied by time, a concept, and a figure in the text, whose voice and vantage point in relation to death furthers the unreliability of the notion of time. These further challenge individuals’ understanding of complex scientific and philosophical views in a variety of ways. The science-fiction genre is essential when considering the speculative nature of It Goes On, which deals with parallel realities and is a fantastical exploration of human ingenuity in plausible futures. Therefore, this paper documents the research-led methodology used to create It Goes On, the application of the ‘many worlds’ theory within a framed narrative, and the many innovative techniques used to contribute new knowledge in a variety of fields.

Keywords: time, many-worlds theory, Heideggerian philosophy, framed narrative

Procedia PDF Downloads 83

Authors: Vidyarini Dwita, Rebecca Fanany

Abstract:

This study will discuss the growth of the market for smartphone technology in Indonesia. This country, with the world’s fourth largest population, has a reputation as the social media capital of the world, and this reputation is largely justified. The penetration of social media is high in Indonesia which has one of the largest global markets. Most Indonesian users of Facebook, Twitter and other social media platforms access the sites from their mobile phones. Indonesia is expected to continue to be a major market for digital mobile devices, such as smartphone and tablets that can access the internet. The aim of this study to describe the way responses of Indonesian consumers to smartphone advertising using English and Indonesian will impact on their perceptions of the prestige and product image of the advertised items and thus influence consumer intention to purchase the item. Advertising for smartphones and similar products is intense and dynamic and often draws on the social attitudes of Indonesians with respect to linguistic and cultural content and especially appeals to their desire to be part of global mainstream culture. The study uses a qualitative method based on in-depth interviews with 30 participants. Content analysis is employed to analyse the responses of Indonesian consumers to smartphone advertising that uses English and Indonesian text. Its findings indicate that consumers’ impressions of English and Indonesian slogans influence their attitudes toward smartphones, suggesting that linguistic context plays a role in influencing consumer purchases.

Keywords: consumer purchases, marketing communication, product image, smartphone advertising, sociolinguistic

Procedia PDF Downloads 224

Authors: Jordana Lenhardt

Abstract:

This paperwork aims to promote a reflection on the critical literacy and multiliteracies concepts in the English language teaching, under an emancipatory perspective, in the English language classroom at the Federal Institute of Mato Grosso (IFMT), Rondonópolis Campus. Some Authors place the relationship between the world conscience and the self-conscience in a direct reason, compromising one to the other, and others defend that emancipatory teaching practice must be connected in all the spheres of the social context; with this paperwork, we intend to analyze students’ interactions with the English language, in order to verify if they demonstrate critical conscience about language and the world around them. The study is still at a preliminary level and is grounded in discourse critical analysis and systemic-functional linguistics. We understand that text is irremediable, linked to a context, and that the linguistic selection made by the speaker builds social representations. This research foresees the analysis of some students’ speeches in an interview about their classes at the Federal Institute in the city of Rondonópolis and the methodology being used on them. Discourse critical analysis explains that, through the awareness of the language uses, learners can become more conscious of the coercions in their own language practices, the possibilities of risks, and the costs of the individual or collective challenges, to engage themselves in emancipatory linguistic practice. The critical language conscience contributes, on the other hand, to make students more aware of the practices in which they are involved, as producers and consumers of texts, of the social forces, ideologies, and power relations, their effects on the identities and social relations, as well as the discourse role in the social and cultural processes.

Keywords: multiliteracies, critical literacy, emancipation, social transformation

Procedia PDF Downloads 101

Authors: Robertgie L. Pianar

Abstract:

Post-colonial theory, post-colonialism, or Poco is a recently established literary theory. Consequently, not many literary works, local and international, have been subjected to its criticism. To help intellectualize local literary texts, in particular, through post-colonial discourse, this qualitative inquiry unfolded. Textual analysis was employed to describe, analyse, and interpret Doms Pagliawan’s Fire Extinguisher, a regional work of literature, grounded on the postcolonial concepts of Edward Said’s Otherness, Homi Bhabha’s Unhomeliness or Paralysis, and Frantz Fanon’s Cultural Resistance. The in-depth reading affirmed that the story contains those postcolonial attributes, revealing the following; (A) the presence of the colonizer, who successfully established colonial control over the colonized, the other, was found; (B) through power superimposition, the colonized character was silenced or paralyzed; and, (C) forms of cultural resistance from the colonized character were shown but no matter how its character avoids ‘postcolonial acts’, the struggle just intensifies, hence inevitable. Pagliawan’s Fire Extinguisher is thus a post-colonial text realizer between two differing cultures, the colonizer and the other. Results of this study may substantiate classroom discussions, both undergraduate and graduate classes, specifically in Philippine and World literature, 21st Century literature, readings in New English literatures, and literary theory and criticism courses, scaffolding learners’ grasp of post-colonialism as a major literary theory drawing classic exemplifications from this regional work.

Keywords: cultural resistance, otherness, post-colonialism, textual analysis, unhomeliness/paralysis

Procedia PDF Downloads 261

Authors: M. Sangeetha, D. Chamundeeswari, C. Saravanababu, C. Rose, V. Gopal

Abstract:

Rheumatoid arthritis (RA) is a chronic, progressive, systemic inflammatory disorder affecting the synovial joints and typically producing symmetrical arthritis that leads to joint destruction, which is responsible for the deformity and disability. Despite improvements in the treatment of RA over the past decade, there still is a need for new therapeutic agents that are efficacious, less expensive, and free of severe adverse reactions. The present study aimed to investigate role of inflammatory markers in arthritic rats treated with ethanolic bark extract of Albizia procera. The protective effect of ethanolic bark extract of Albizia procera against complete Freund’s adjuvant (CFA) induced arthritis in rats. Arthritis was induced by an intradermal injection of 0.1 ml FCA in the foot pad of left hind limb of rats. ETBE (100 and 200 mg/kg b.wt./p.o) and the reference drug diclofenac (25 mg/kg b.wt./p.o) were administered to arthritic rats. Paw volume was measured for all the animals before inducing arthritis and thereafter once in seven days by using plethysmometer for 42 days. Gene expression of inflammatory markers such as IL-1β and IL-10 were investigated in paw tissues. Up regulation of IL-1β and Down regulation IL-10 were observed in CFA injected rats when compared to normal rats. ETBE attenuated these alterations dose dependently when compared to the vehicle treated rats. These results provide insights into the mechanism of anti-arthritic activity, and unravel potential therapeutic use of Albizia procera in arthritis.

Keywords: CFA-Complete Freund’s adjuvant, ETBE – ethanolic bark extract, IL- interleukins, RA-rheumatoid arthritis

Procedia PDF Downloads 285

Authors: Samaneh Khalili

Abstract:

Natural law theory, in moral philosophy, refers to a system of unchanging values held to be mutual to all humans and can be discovered through reason. The natural law theory is commonly associated with western Philosophers. In contrast, discussions on notions of natural law in Islamic intellectual history were relatively rare. This paper aims to show that the moral theory developed by the Mu'tazilite thinkers can be classified in the ideas of natural law. In doing so, this study will demonstrate that the objective and unchanging values, according to Mu'tazilite theologians, provide the guidelines for assessing the Islamic law rules in the field of human coexistence. The focus of the paper lies on ʿAbd al-Ğabbār, who was the most influential thinker in the late epoch of the Muʿtazila. Although ʿAbd al-Ǧabbār did not leave a text with a systematic discussion of natural law, his teaching of nature, human reason, and the moral values of actions are all scattered throughout his work -'al-Muġnī fī abwāb at-tawḥīd wa-l-'adl'. It is necessary to focus on ʿAbd al-Ǧabbār's theories on reason, nature, and ethics since natural law revolves around the basic concepts of nature, reason, and moral value. While analyzing the concept of the Nature, it will attempt to answer how he explains the world's physical structure and God's relationship to natural events. Moreover, from ʿAbd al-Ǧabbār's point of view, is nature a self-determined system that follows its inner principle in every kind of change, or is nature guided by an external power? Does causality govern natural events? About the concept of reason, an attempt is made to examine how human reason, according to ʿAbd al-Ǧabbār, conceives moral attributes. Finally, the Autor will discuss the concepts of objective values and the place of rights and duties derived from Islamic law in ʿAbd al-Ǧabbār's thought.

Keywords: Islamic law, Mu'tazilite theology, natural law in Islamic theology, objective and unchanging values.

Procedia PDF Downloads 96

Authors: Marie Komorna, Katerina Hadkova

Abstract:

One reason why hearing disabilities as compared to other disabilities are considered to be less serious, is the belief that deaf and hard of hearing persons can read and write without problems and can therefore fairly easily compensate for problems related to their limited ability to hear sound. However in reality this is not the case, especially as regards written Czech, deaf persons are often not able to communicate their message clearly to its recipients. Their inability to communicate fully in written language is one of the most severe problems facing a number of deaf persons, a problem which they face and which makes it difficult for them to function in a sound-based environment. Despite this fact, this issue is one which has been given only a minimum of attention in the Czech Republic. That is why we decided to focus our research on this issue, specifically targeting written communication of deaf pupils in primary and secondary schools. The paper summarizes the background and objectives of this research. The written work of deaf respondents was obtained in response to a narrative based on a series of images which depicted a continuous storyline. Based on an analysis of the obtained written work we tried to describe the specifics of the narrative abilities of the deaf authors of these texts. We also analyzed other aspects and specific traits of text written by deaf authors at a phonetic-phonological, lexical-semantic, morphological and syntactic, respectively pragmatic level. Based on the results of the project it will be possible to increase knowledge of the communication abilities of deaf persons in written Czech. The obtained data may be used during future research and for teaching purposes and/or education concepts for teaching Czech to deaf pupils.

Keywords: communication competence, deaf, narrative, written texts

Procedia PDF Downloads 338

Authors: Parthiban Srinivasan

Abstract:

With recent advances in computational power and access to enough data in biosciences, artificial intelligence methods are increasingly being used in drug discovery research. These methods are essentially a series of advanced statistics based exercises that review the past to indicate the likely future. Our goal is to develop a model that accurately predicts biological activity and toxicity parameters for novel compounds. We have compiled a robust library of over 150,000 chemical compounds with different pharmacological properties from literature and public domain databases. The compounds are stored in simplified molecular-input line-entry system (SMILES), a commonly used text encoding for organic molecules. We utilize an automated process to generate an array of numerical descriptors (features) for each molecule. Redundant and irrelevant descriptors are eliminated iteratively. Our prediction engine is based on a portfolio of machine learning algorithms. We found Random Forest algorithm to be a better choice for this analysis. We captured non-linear relationship in the data and formed a prediction model with reasonable accuracy by averaging across a large number of randomized decision trees. Our next step is to apply deep neural network (DNN) algorithm to predict the biological activity and toxicity properties. We expect the DNN algorithm to give better results and improve the accuracy of the prediction. This presentation will review all these prominent machine learning and deep learning methods, our implementation protocols and discuss these techniques for their usefulness in biomedical and health informatics.

Keywords: deep learning, drug discovery, health informatics, machine learning, toxicity prediction

Procedia PDF Downloads 356

Authors: Macky Kato, Takeshi Sato, Mizuki Nakajima

Abstract:

One major problem related to the use of smartphones is texting while simultaneously engaging in other things, resulting in serious road accidents. Apart from texting while driving being one of the most dangerous behaviors, texting while walking is also dangerous because it narrows the pedestrians’ field of vision. However, many of pedestrian text while walking very habitually. Smartphone users often overlook the potential harm associated with this behavior even while crossing roads. The successful texting while walking make them think that they are safe. The purpose of this study is to reveal of the influence of peripheral vision to the stability of walking trajectory with texting while walking. In total, 9 healthy male university students participated in the experiment. Their mean age was 21.4 years, and standard deviation was 0.7 years. They attempted to walk 10 m in three conditions. First one is the control (CTR) condition, with no phone and no restriction. The second one is the texting while walking (TWG) with no restrictions. The third one is restriction condition (PRS), with phone restricted by experimental peripheral goggles. The horizontal distances (HDS) and directions are measured as the scale of horizontal stability. The longitudinal distances (LDS) between the footprints were measured as the scale of the walking rhythm. The results showed that the HDS of the footprints from the straight line increased as the participants walked in the TWG and PRS conditions. In the PRS condition, this tendency was particularly remarkable. In addition, the LDS between the footprints decreased in the order of the CTR, TWG, and PRS conditions. The ANOVA results showed significant differences in the three conditions with respect to HDS. The differences among these conditions showed that the narrowing of the Pedestrian's vision because of smartphone use influences the walking trajectory and rhythm. It can be said that the pedestrians seem to use their peripheral vision marginally on texting while walking. Therefore, we concluded that the texting while walking narrows the peripheral vision so danger to increase the risk of the accidents.

Keywords: peripheral vision, stability, texting while walking, walking trajectory

Procedia PDF Downloads 257

Authors: Xianghong Lu, Yuejun Wang, Chengshi Gan, Xin Qian

Abstract:

The North Sulawesi Arc, located on the east of Indonesia and to the south of the Celebes Sea, is the north part of the K-shape of Sulawesi Island and has a complex tectonic history since the Cenozoic due to the convergence of three plates (Eurasia, India-Australia and Pacific plates). Published rock records contain less precise chronology, mostly using K-Ar dating, and rare geochemistry data, which limit the understanding of the regional tectonic setting. This study presents detailed zircon U-Pb geochronological and Hf-O isotope and whole-rock geochemical analyses for the Na-rich granites from the North Sulawesi Arc. Zircon U-Pb geochronological analyses of three representative samples yield weighted mean ages of 30.4 ± 0.4 Ma, 29.5 ± 0.2 Ma, and 27.3 ± 0.4 Ma, respectively, revealing the Oligocene magmatism in the North Sulawesi Arc. The samples have high Na₂O and low K₂O contents with high Na₂O/K₂O ratios, belonging to Low-K tholeiitic Na-rich granites. The Na-rich granites are characterized by high SiO₂ contents (75.05-79.38 wt.%) and low MgO contents (0.07-0.91 wt.%) and show arc-like trace elemental signatures. They have low (⁸⁷Sr/⁸⁶Sr)i ratios (0.7044-0.7046), high εNd(t) values (from +5.1 to +6.6), high zircon εHf(t) values (from +10.1 to +18.8) and low zircon δ18O values (3.65-5.02). They show an Indian-Ocean affinity of Pb isotopic compositions with ²⁰⁶Pb/²⁰⁴Pb ratio of 18.16-18.37, ²⁰⁷Pb/²⁰⁴Pb ratio of 15.56-15.62, and ²⁰⁸Pb/²⁰⁴Pb ratio of 38.20-38.66. These geochemical signatures suggest that the Oligocene Na-rich granites from the North Sulawesi Arc formed by partial melting of the juvenile oceanic crust with sediment-derived fluid-related metasomatism in a subducting setting and support an intra-oceanic arc origin. Combined with the published study, the emergence of extensive calc-alkaline felsic arc magmatism can be traced back to the Early Oligocene period, subsequent to the Eocene back-arc basalts (BAB) that share similarity with the Celebes Sea basement. Since the opening of the Celebes Sea started from the Eocene (42~47 Ma) and stopped by the Early Oligocene (~32 Ma), the geodynamical mechanism of the formation of the Na-rich granites from the North Sulawesi Arc during the Oligocene might relate to the subduction of the Indian Ocean.

Keywords: North Sulawesi Arc, oligocene, Na-rich granites, in-situ zircon Hf–O analysis, intra-oceanic origin

Procedia PDF Downloads 76

Authors: Yosep Chong, Yejin Kim, Jingyun Choi, Hwanjo Yu, Eun Jung Lee, Chang Suk Kang

Abstract:

For the past decades, immunohistochemistry (IHC) has been playing an important role in the diagnosis of human neoplasms, by helping pathologists to make a clearer decision on differential diagnosis, subtyping, personalized treatment plan, and finally prognosis prediction. However, the IHC performed in various tumors of daily practice often shows conflicting and very challenging results to interpret. Even comprehensive diagnosis synthesizing clinical, histologic and immunohistochemical findings can be helpless in some twisted cases. Another important issue is that the IHC data is increasing exponentially and more and more information have to be taken into account. For this reason, we reached an idea to develop an expert supporting system to help pathologists to make a better decision in diagnosing human neoplasms with IHC results. We gave probabilistic decision tree algorithm and tested the algorithm with real case data of lymphoid neoplasms, in which the IHC profile is more important to make a proper diagnosis than other human neoplasms. We designed probabilistic decision tree based on Bayesian theorem, program computational process using MATLAB (The MathWorks, Inc., USA) and prepared IHC profile database (about 104 disease category and 88 IHC antibodies) based on WHO classification by reviewing the literature. The initial probability of each neoplasm was set with the epidemiologic data of lymphoid neoplasm in Korea. With the IHC results of 131 patients sequentially selected, top three presumptive diagnoses for each case were made and compared with the original diagnoses. After the review of the data, 124 out of 131 were used for final analysis. As a result, the presumptive diagnoses were concordant with the original diagnoses in 118 cases (93.7%). The major reason of discordant cases was that the similarity of the IHC profile between two or three different neoplasms. The expert supporting system algorithm presented in this study is in its elementary stage and need more optimization using more advanced technology such as deep-learning with data of real cases, especially in differentiating T-cell lymphomas. Although it needs more refinement, it may be used to aid pathological decision making in future. A further application to determine IHC antibodies for a certain subset of differential diagnoses might be possible in near future.

Keywords: database, expert supporting system, immunohistochemistry, probabilistic decision tree

Procedia PDF Downloads 224

Authors: Meaghan Bowman

Abstract:

Soldiers are taught about the importance of effective communication with repetition of the phrase, “Communication is key.” They receive training in preparing for, and carrying out, interactions between foreign and domestic leaders to gain crucial information about a mission. These interactions are known as Key Leader Engagements (KLEs). For the training of KLEs, doctrine mandates the skills needed to conduct these “engagements” such as how to: behave appropriately, identify key leaders, and employ effective strategies. Army officers in training learn how to confront leaders, what information to gain, and how to ask questions respectfully. Unfortunately, soldiers rarely learn how to formulate questions optimally. Since less complex questions are easier to understand, we hypothesize that semantic complexity affects content understanding, and that age and education levels may have an effect on one’s ability to form paraphrases and judge their quality. In this study, we looked at paraphrases of queries as well as judgments of both the paraphrases’ naturalness and their semantic similarity to the query. Queries were divided into three complexity categories based on the number of relations (the first number) and the number of knowledge graph edges (the second number). Two crowd-sourced tasks were completed by Amazon volunteer participants, also known as turkers, to answer the research questions: (i) Are more complex queries harder to paraphrase and judge and (ii) Do age and education level affect the ability to understand complex queries. We ran statistical tests as follows: MANOVA for query understanding and two-way ANOVA to understand the relationship between query complexity and education and age. A probe of the number of given-level queries selected for paraphrasing by crowd-sourced workers in seven age ranges yielded promising results. We found significant evidence that age plays a role and marginally significant evidence that education level plays a role. These preliminary tests, with output p-values of 0.0002 and 0.068, respectively, suggest the importance of content understanding in a communication skill set. This basic ability to communicate, which may differ by age and education, permits reproduction and quality assessment and is crucial in training soldiers for effective participation in KLEs.

Keywords: engagement, key leader, paraphrasing, query complexity, understanding

Procedia PDF Downloads 161

Authors: Shahnam Behnam Malekzadeh, Ian Kerr, Tyson Kaempffer, Teague Harper, Andrew Watson

Abstract:

The Site C Hydroelectric dam is currently being constructed in north-eastern British Columbia on sub-horizontal sedimentary strata that dip approximately 15 meters from one bank of the Peace River to the other. More than 615 pressure sensors (Vibrating Wire Piezometers) have been installed on bedding planes (BPs) since construction began, with over 80 more planned before project completion. These pressure measurements are essential to monitor the stability of the rock foundation during and after construction and for dam safety purposes. BPs are identified by their clay gouge infilling, which varies in thickness from less than 1 to 20 mm and can be challenging to identify as the core drilling process often disturbs or washes away the gouge material. Without the use of depth predictions from nearby boreholes, stratigraphic markers, and downhole geophysical data, it is difficult to confidently identify BP targets for the sensors. In this paper, a Case-Based Reasoning (CBR) method was used to develop an empirical model called the Bedding Plane Elevation Prediction (BPEP) to help geologists and geotechnical engineers to predict geological features and bedding planes at new locations in a fast and accurate manner. To develop CBR, a database was developed based on 64 pressure sensors already installed on key bedding planes BP25, BP28, and BP31 on the Right Bank, including bedding plane elevations and coordinates. Thirteen (20%) of the most recent cases were selected to validate and evaluate the accuracy of the developed model, while the similarity was defined as the distance between previous cases and recent cases to predict the depth of significant BPs. The average difference between actual BP elevations and predicted elevations for above BPs was ±55cm, while the actual results showed that 69% of predicted elevations were within ±79 cm of actual BP elevations while 100% of predicted elevations for new cases were within ±99cm range. Eventually, the actual results will be used to develop the database and improve BPEP to perform as a learning machine to predict more accurate BP elevations for future sensor installations.

Keywords: case-based reasoning, geological feature, geology, piezometer, pressure sensor, core logging, dam construction

Procedia PDF Downloads 79

Authors: Mohd Noah A. Rahman, Afzaal H. Seyal, Sharul T. Tajuddin, Hartiny Md Azmi

Abstract:

Taking into consideration the real time situation specifically the higher academic institutions, small, medium to large companies, public to private sectors and the remaining sectors, do experience the inventory or asset shrinkages due to theft, loss or even inventory tracking errors. This happening is due to a zero or poor security systems and measures being taken and implemented in their organizations. Henceforth, implementing the Radio Frequency Identification (RFID) technology into any manual or existing web-based system or web application can simply deter and will eventually solve certain major issues to serve better data retrieval and data access. Having said, this manual or existing system can be enhanced into a mobile-based system or application. In addition to that, the availability of internet connections can aid better services of the system. Such involvement of various technologies resulting various privileges to individuals or organizations in terms of accessibility, availability, mobility, efficiency, effectiveness, real-time information and also security. This paper will look deeper into the integration of mobile devices with RFID technologies with the purpose of asset tracking and control. Next, it is to be followed by the development and utilization of MongoDB as the main database to store data and its association with RFID technology. Finally, the development of a web based system which can be viewed in a mobile based formation with the aid of Hypertext Preprocessor (PHP), MongoDB, Hyper-Text Markup Language 5 (HTML5), Android, JavaScript and AJAX programming language.

Keywords: RFID, asset tracking system, MongoDB, NoSQL

Procedia PDF Downloads 306

Authors: Jude K. Safo

Abstract:

Knowledge Graphs KG) and their relation to Graph Embeddings (GE) represent a unique data structure in the landscape of machine learning (relative to image, text and acoustic data). Unlike the latter, GEs are the only data structure sufficient for representing hierarchically dense, semantic information needed for use-cases like supply chain data and protein folding where the search space exceeds the limits traditional search methods (e.g. page-rank, Dijkstra, etc.). While GEs are effective for compressing low rank tensor data, at scale, they begin to introduce a new problem of ’data retreival’ which we observe in Large Language Models. Notable attempts by transE, TransR and other prominent industry standards have shown a peak performance just north of 57% on WN18 and FB15K benchmarks, insufficient practical industry applications. They’re also limited, in scope, to next node/link predictions. Traditional linear methods like Tucker, CP, PARAFAC and CANDECOMP quickly hit memory limits on tensors exceeding 6.4 million nodes. This paper outlines a topological framework for linear mapping between concepts in KG space and GE space that preserve cardinality. Most importantly we introduce a traceable framework for composing dense linguistic strcutures. We demonstrate performance on WN18 benchmark this model hits. This model does not rely on Large Langauge Models (LLM) though the applications are certainy relevant here as well.

Keywords: representation theory, large language models, graph embeddings, applied algebraic topology, applied knot theory, combinatorics

Procedia PDF Downloads 68

Authors: Asma Rashid, Shazia Iram, Iftikhar Ahmad

Abstract:

Mango sudden death is an emerging problem in Pakistan. As its prevalence is observed in almost all mango growing areas and severity varied from 2-5% in Punjab and 5-10% in Sindh. Symptoms on affected trees include bark splitting, discoloration of the vascular tissue, wilting, gummosis and at the end rapid death. Total of n= 45 isolates were isolated from different mango growing areas of Punjab and Sindh. Pathogenicity of these fungal isolates was tested through artificial inoculation method on different hosts (potato tubers, detached mango leaves, detached mango twigs and mango plants) under controlled conditions and all were proved pathogenic with varying degree of aggressiveness in reference to control. The findings of the present study proved that out of these four methods, potato tubers inoculation method was the most ideal as this fix the inoculums on the target site. Increased fungal growth and spore numbers may be due to soft tissues of potato tubers from which Ceratocystis isolates can easily pass. Lesion area on potato tubers was in the range of 7.09-0.14 cm2 followed by detached mango twigs which were ranged from 0.48-0.09 cm2). All pathological results were proved highly significant at P<0.05 through ANOVA but isolate to isolate showed non-significant behaviour but they have the positive effect on lesion area. Re-isolation of respective fungi was achieved with 100 percent success which results in the verification of Koch’s postulates. DNA of fungal pathogens was successfully extracted through phenol chloroform method. Amplification was done through ITS, b-tubulin gene, and Transcription Elongation Factor (EF1-a) gene primers and the amplified amplicons were sequenced and compared from NCBI which showed 99-100 % similarity with Ceratocystis manginecans. Fungus Ceratocystis manginecans formed one of strongly supported sub-clades through phylogenetic tree. Results obtained through this work would be supportive in establishment of relation of isolates with their region and will give information about pathogenicity level of isolates that would be useful to develop the management policies to reduce the afflictions in orchards caused by mango sudden death.

Keywords: artificial inoculation, mango, Ceratocystis manginecans, phylogenetic, screening

Procedia PDF Downloads 248

Authors: S. Fröhlich, M. Herold, M. Allmer

Abstract:

Early stage quantitative analysis of host cell protein (HCP) variations is challenging yet necessary for comprehensive bioprocess development. High resolution mass spectrometry (HRMS) provides a high-end technology for accurate identification alongside with quantitative information. Hereby we describe a flexible HRMS assay platform to quantify HCPs relevant in microbial expression systems such as E. Coli in both up and downstream development by means of MVDA tools. Cell pellets were lysed and proteins extracted, purified samples not further treated before applying the SMART tryptic digest kit. Peptides separation was optimized using an RP-UHPLC separation platform. HRMS-MSMS analysis was conducted on an Orbitrap Velos Elite applying CID. Quantification was performed label-free taking into account ionization properties and physicochemical peptide similarities. Results were analyzed using SIEVE 2.0 (Thermo Fisher Scientific) and SIMCA (Umetrics AG). The developed HRMS platform was applied to an E. Coli expression set with varying productivity and the corresponding downstream process. Selected HCPs were successfully quantified within the fmol range. Analysing HCP networks based on pattern analysis facilitated low level quantification and enhanced validity. This approach is of high relevance for high-throughput screening experiments during upstream development, e.g. for titer determination, dynamic HCP network analysis or product characterization. Considering the downstream purification process, physicochemical clustering of identified HCPs is of relevance to adjust buffer conditions accordingly. However, the technology provides an innovative approach for label-free MS based quantification relying on statistical pattern analysis and comparison. Absolute quantification based on physicochemical properties and peptide similarity score provides a technological approach without the need of sophisticated sample preparation strategies and is therefore proven to be straightforward, sensitive and highly reproducible in terms of product characterization.

Keywords: process analytical technology, mass spectrometry, process characterization, MVDA, pattern recognition

Procedia PDF Downloads 249