Search results for: gene expression profiles

Authors: Hassan Sabo

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Language is a system of conventionally spoken, manual and written symbols by human beings that members of a certain social group and participants in its culture express themselves. Communication, expression of identity and imaginative expression are among the functions of language. Dialect is a form of language, or a regional variety of language that is spoken in a particular geographical setting by a particular group of people. Hausa is one of the major languages in Africa, in terms of large number of people for whom it is the first language. Hausa is one of the western Chadic groups of languages. It constitutes one of the five or six branches of Afro-Asiatic family. The predominant Hausa speakers are in Nigeria and they live in different geographical locations which resulted to variety of dialects within the Hausa language apart of the standard Hausa language, the Hausa language has a variety of dialect that distinguish from one another by such features as phonology, grammar and vocabulary. This study intends to examine such features that serve as means of identification among Hausa speakers who are set off from others, geographically or socially.

Keywords: dialect, features, geographical location, Hausa language

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Authors: I. Boroňová, J.Bernasovská, J. Kľoc, Z. Tomková, E. Petrejčíková, D. Gabriková, S. Mačeková

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Osteoporosis is a complex health disease characterized by low bone mineral density, which is determined by an interaction of genetics with metabolic and environmental factors. Current research in genetics of osteoporosis is focused on identification of responsible genes and polymorphisms. TNFRSF11B gene plays a key role in bone remodeling. The aim of this study was to investigate the genotype and allele distribution of A163G (rs3102735) osteoprotegerin gene promoter and G1181C (rs2073618) osteoprotegerin first exon polymorphisms in the group of 180 unrelated postmenopausal women with diagnosed osteoporosis and 180 normal controls. Genomic DNA was isolated from peripheral blood leukocytes using standard methodology. Genotyping for presence of different polymorphisms was performed using the Custom Taqman®SNP Genotyping assays. Hardy-Weinberg equilibrium was tested for each SNP in the groups of participants using the chi-square (χ2) test. The distribution of investigated genotypes in the group of patients with osteoporosis were as follows: AA (66.7%), AG (32.2%), GG (1.1%) for A163G polymorphism; GG (19.4%), CG (44.4%), CC (36.1%) for G1181C polymorphism. The distribution of genotypes in normal controls were follows: AA (71.1%), AG (26.1%), GG (2.8%) for A163G polymorphism; GG (22.2%), CG (48.9%), CC (28.9%) for G1181C polymorphism. In A163G polymorphism the variant G allele was more common among patients with osteoporosis: 17.2% versus 15.8% in normal controls. Also, in G1181C polymorphism the phenomenon of more frequent occurrence of C allele in the group of patients with osteoporosis was observed (58.3% versus 53.3%). Genotype and allele distributions showed no significant differences (A163G: χ2=0.270, p=0.605; χ2=0.250, p=0.616; G1181C: χ2= 1.730, p=0.188; χ2=1.820, p=0.177). Our results represents an initial study, further studies of more numerous file and associations studies will be carried out. Knowing the distribution of genotypes is important for assessing the impact of these polymorphisms on various parameters associated with osteoporosis. Screening for identification of “at-risk” women likely to develop osteoporosis and initiating subsequent early intervention appears to be most effective strategy to substantially reduce the risks of osteoporosis.

Keywords: osteoporosis, real-time PCR method, SNP polymorphisms

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Authors: Van-Dycke Sarpong Asare, Vincent Adongo

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Field measurement of subsurface seismic p-wave velocities was undertaken through seismic refraction tomography. The aim of this work is to obtain a model of the shallow subsurface material elastic properties relevant for geotechnical site characterization. The survey area is at Pwalugu in Northern Ghana, where a multipurpose dam, for electricity generation, irrigation, and potable water delivery, is being planned. A 24-channel seismograph and 24, 10 Hz electromagnetic geophones, deployed 5 m apart constituted the acquisition hardware. Eleven (2-D) seismic refraction profiles, nine of which ran almost perpendicular and two parallel to the White Volta at Pwalugu, were acquired. The refraction tomograms of the thirteen profiles revealed a subsurface model consisting of one minor and one major acoustic impedance boundaries – the top dry/loose sand and the variably weathered sandstone contact, and the overburden-sandstones bedrock contact respectively. The p-wave velocities and by inference, with a priori values of poison ratios, the s-wave velocities, assisted in characterizing the geotechnical conditions of the proposed site and also in evaluating the dynamic properties such as the maximum shear modulus, the bulk modulus, and the Young modulus.

Keywords: tomography, characterization, consolidated, Pwalugu and seismograph

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Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

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Authors: M. Caruffo, P. Navarrete, C. G. Feijoo, L. Sáenz

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Disease prevention through the use of vaccines has been crucial to achieve the current level of production in the salmon industry. However, vaccines have been developed based largely on inactivated bacterial formulations, using the whole pathogen. These formulations have demonstrated excellent efficacy against extracellular bacterial pathogens. However diseases with the greatest economic impacts correspond to intracellular bacterial and viral pathogens, vaccines based on these types of agents have shown a discrete effectiveness. It is for these reasons that the development of subunit vaccines based on defined antigens offers a promising solution. The main problem is that subunit vaccines offer a low immunogenicity, since they lack immunostimulatory elements, so that the development of new adjuvants platforms becomes an important challenge for this type of formulations. We evaluate the effect of a formulation based on proteoliposomes of Vibrio anguillarum administered by immersion as a new adjuvant strategy, allowing efficient stimulation of the innate immune system. Proteoliposomes physicochemical properties were evaluated in its ability to produce an inflammatory process. Using zebrafish (Danio rerio) larvae as a model species and the transgenic line (Tg(mpx: GFP)i114) allowed us to track the neutrophil migration in real time. Additionally we evaluated the gene expression of some molecular markers involved in the development of the innate immune response characterizing the adjuvant capacity of the formulation.

Keywords: adjuvants, vaccine development, zebrafish, innate immunity

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Authors: András Farkas, István Molnár, Jan Vrána, Veronika Burešová, Petr Cápal, András Cseh, Márta Molnár-Láng, Jaroslav Doležel

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Species of Aegilops played a central role in the evolution of wheat and are sources of traits related to yield quality and tolerance against biotic and abiotic stresses. These wild genes and alleles are desirable to use in crop improvement programs via introgressive hybridization. However, the success of chromosome mediated gene transfer to wheat are hampered by the pour knowledge on the genome structure of Aegilops relative to wheat and by the low number of cost-effective molecular markers specific for Aegilops chromosomes. The COS markers specific for genes conserved throughout evolution in both sequence and copy number between Triticeae/Aegilops taxa and define orthologous regions, thus enabling the comparison of regions on the chromosomes of related species. The present study compared individual chromosomes of Aegilops umbellulata (UU), Ae. comosa (MM), Ae. speltoides (SS) and Ae. caudata (CC) purified by flourescent labelling with oligonucleotid SSR repeats and biparametric flow cytometry with wheat by identifying orthologous chromosomal regions by COS markers. The linear order of bin-mapped COS markers along the wheat D chromosomes was identified by the use of chromosome-specific sequence data and virtual gene order. Syntenic regions of wheat identifying genome rearrangements differentiating the U, M, S or C genomes from the D genome of wheat were detected. The conserved orthologous set markers assigned to Aegilops chromosomes promise to accelerate gene introgression by facilitating the identification of alien chromatin. The syntenic relationships between the Aegilops species and wheat will facilitate the targeted development of new markers specific for U, M, S and C genomic regions and will contribute to the understanding of molecular processes related to the evolution of Aegilops.

Keywords: Aegilops, cos-markers, flow-sorting, wheat

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Authors: Hend K. Moosa, Eman A. Rashwan, Ezzat M. Hassan, Amany A. Ghazy, Amel G. Sheredy

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The stability of microRNA (miR) in the circulation can show a great progress toward the discovery of non-invasive diagnostic and prognostic biomarkers in many diseases. In the present study, circulatory miR-122 and miR-130a were analysed in chronic hepatitis C Egyptian patients in predicting the clinical outcome of interferon treatment. In addition, their expression levels were correlated to viral RNA levels, necro-inflammatory markers (AST, ALT) and to each other. This study was conducted on 51 subjects where 36 were chronic HCV patients in which they were divided into naive and interferon treated HCV patients (responders and non-responders) and 15 matched healthy controls. Serum quantification of miR-122 and miR-130a were performed by quantitative Real-time Polymerase Chain Reaction (qRT-PCR). The results showed a significant upregulation of miR-122 in non-responder patients (P=0.049). By receiver operating characteristic analysis curve, miR-122 revealed 65% sensitivity and 92.3% specificity in predicting non-responsiveness of patients to IFN treatment, while miR-130a showed a sensitivity of 100% and specificity of 53.85%. Remarkably, there was a significant positive correlation between miR-122 and miR-130a in naive HCV patients (r=0.714, p=0.003). However, there was no significant correlation between serum miR-122, miR-130a expression levels and necro-inflammatory markers (AST, ALT). To conclude, miR-122 and miR-130a have a significant association with viral RNA levels and accordingly, they may have a synergistic power in promoting viral replication. Interestingly, miR-122 and miR-130a have a predictive power in predicting clinical outcome of IFN treatment which can be further studied in currently used drugs in order to reduce the socio-economic burden of potentially non-responders.

Keywords: hepatitis C, microRNA, miR-122, miR-130a

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Authors: Ali Rashash Alzahrani, Elizabeth Stojanovski

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Data on various aspects of education are collected at the institutional and government level regularly. In Australia, for example, students at various levels of schooling undertake examinations in numeracy and literacy as part of NAPLAN testing, enabling longitudinal assessment of such data as well as comparisons between schools and states within Australia. Another source of educational data collected internationally is via the PISA study which collects data from several countries when students are approximately 15 years of age and enables comparisons in the performance of science, mathematics and English between countries as well as ranking of countries based on performance in these standardised tests. As well as student and school outcomes based on the tests taken as part of the PISA study, there is a wealth of other data collected in the study including parental demographics data and data related to teaching strategies used by educators. Overall, an abundance of educational data is available which has the potential to be used to help improve educational attainment and teaching of content in order to improve learning outcomes. A multivariate assessment of such data enables multiple variables to be considered simultaneously and will be used in the present study to help develop profiles of students based on performance in mathematics using data obtained from the PISA study.

Keywords: cluster analysis, education, mathematics, profiles

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Authors: Ge Zheng, Jun Peng

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Unbalanced power output from individual cells of Proton Exchange Membrane Fuel Cell (PEMFC) has direct effects on PEMFC stack performance, in particular under transient operation. In the paper, a multi-layer ANN (Artificial Neural Network) model Radial Basis Functions (RBF) has been developed for predicting cells' output profiles by applying gas supply parameters, cooling conditions, temperature measurement of individual cells, etc. The feed-forward ANN model was validated with experimental data. Influence of relevant parameters of RBF on the network accuracy was investigated. After adequate model training, the modelling results show good correspondence between actual measurements and reconstructed output profiles. Finally, after the model was used to optimize the stack output performance under steady-state and transient operating conditions, it suggested that the developed ANN control model can help PEMFC stack to have obvious improvement on power output under fast acceleration process.

Keywords: proton exchange membrane fuel cell, PEMFC, artificial neural network, ANN, cell output profile, transient

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Authors: Ksheeraj Sai Vepuri, Nada Attar

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We as humans use words with accompanying visual and facial cues to communicate effectively. Classifying facial emotion using computer vision methodologies has been an active research area in the computer vision field. In this paper, we propose a simple method for facial expression recognition that enhances accuracy. We tested our method on the FER-2013 dataset that contains static images. Instead of using Histogram equalization to preprocess the dataset, we used Unsharp Mask to emphasize texture and details and sharpened the edges. We also used ImageDataGenerator from Keras library for data augmentation. Then we used Convolutional Neural Networks (CNN) model to classify the images into 7 different facial expressions, yielding an accuracy of 69.46% on the test set. Our results show that using image preprocessing such as the sharpening technique for a CNN model can improve the performance, even when the CNN model is relatively simple.

Keywords: facial expression recognittion, image preprocessing, deep learning, CNN

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Authors: H. G. C. P. Dinesh, G. Tharshini, M. P. B. Ekanayake, G. M. R. I. Godaliyadda

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This paper presents a new spatial feature extraction method based on principle component analysis (PCA) and Fisher Discernment Analysis (FDA) for facial expression recognition. It not only extracts reliable features for classification, but also reduces the feature space dimensions of pattern samples. In this method, first each gray scale image is considered in its entirety as the measurement matrix. Then, principle components (PCs) of row vectors of this matrix and variance of these row vectors along PCs are estimated. Therefore, this method would ensure the preservation of spatial information of the facial image. Afterwards, by incorporating the spectral information of the eigen-filters derived from the PCs, a feature vector was constructed, for a given image. Finally, FDA was used to define a set of basis in a reduced dimension subspace such that the optimal clustering is achieved. The method of FDA defines an inter-class scatter matrix and intra-class scatter matrix to enhance the compactness of each cluster while maximizing the distance between cluster marginal points. In order to matching the test image with the training set, a cosine similarity based Bayesian classification was used. The proposed method was tested on the Cohn-Kanade database and JAFFE database. It was observed that the proposed method which incorporates spatial information to construct an optimal feature space outperforms the standard PCA and FDA based methods.

Keywords: facial expression recognition, principle component analysis (PCA), fisher discernment analysis (FDA), eigen-filter, cosine similarity, bayesian classifier, f-measure

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Authors: Kung-Jen Tu, Danny Vernatha

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To assist individual departments within universities in their energy management tasks, this study explores the application of Building Information Modeling in establishing the ‘BIM based Energy Management Support System’ (BIM-EMSS). The BIM-EMSS consists of six components: (1) sensors installed for each occupant and each equipment, (2) electricity sub-meters (constantly logging lighting, HVAC, and socket electricity consumptions of each room), (3) BIM models of all rooms within individual departments’ facilities, (4) data warehouse (for storing occupancy status and logged electricity consumption data), (5) building energy management system that provides energy managers with various energy management functions, and (6) energy simulation tool (such as eQuest) that generates real time 'standard energy consumptions' data against which 'actual energy consumptions' data are compared and energy efficiency evaluated. Through the building energy management system, the energy manager is able to (a) have 3D visualization (BIM model) of each room, in which the occupancy and equipment status detected by the sensors and the electricity consumptions data logged are displayed constantly; (b) perform real time energy consumption analysis to compare the actual and standard energy consumption profiles of a space; (c) obtain energy consumption anomaly detection warnings on certain rooms so that energy management corrective actions can be further taken (data mining technique is employed to analyze the relation between space occupancy pattern with current space equipment setting to indicate an anomaly, such as when appliances turn on without occupancy); and (d) perform historical energy consumption analysis to review monthly and annually energy consumption profiles and compare them against historical energy profiles. The BIM-EMSS was further implemented in a research lab in the Department of Architecture of NTUST in Taiwan and implementation results presented to illustrate how it can be used to assist individual departments within universities in their energy management tasks.

Keywords: database, electricity sub-meters, energy anomaly detection, sensor

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Authors: Patsy Tan, Dana Baltutis

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This presentation demonstrates the effectiveness of music therapy in co-treatment with speech pathology and occupational therapy as an innovative way when working with children and adolescents with complex individual differences to facilitate communication, emotional, motor and social skills development. Each child with special needs and their carer has an individual profile which encompasses their visual-spatial, auditory, language, learning, mental health, family dynamic, sensory-motor, motor planning and sequencing profiles. The most common issues among children with special needs, especially those diagnosed with Autism Spectrum Disorder, are in the areas of regulation, communication, and social-emotional development. The ability of children living with challenges to communicate and use language and understand verbal and non-verbal information, as well as move their bodies to explore and interact with their environments in social situations, depends on the children being regulated both internally and externally and trusting their communication partners and understanding what is happening in the moment. For carers, it is about understanding the tempo, rhythm, pacing, and timing of their own individual profile, as well as the profile of the child they are interacting with, and how these can sync together. In this study, music therapy is used in co-treatment sessions with a speech pathologist and/or an occupational therapist using the DIRFloortime approach to facilitate the regulation, attention, engagement, reciprocity and social-emotional capacities of children presenting with complex individual differences. Documented changes in 10 domains of children’s development over a 12-month period using the Individual Music Therapy Assessment Profile (IMTAP) were observed. Children were assessed biannually, and results show significant improvements in the social-emotional, musicality and receptive language domains indicating that co-treatment with a music therapist using the DIRFloortime framework is highly effective. This presentation will highlight strategies that facilitate regulation, social-emotional and communication development for children and adolescents with complex individual profiles.

Keywords: communication, shared attention, regulation, social emotional

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Authors: Sahar Javadi-Novashnagh, Mohammad Moradi-Shahrbabak, Mostafa Sadeghi, Katarzyna Ropka-Molik, Hossein Moradi-Shahrbabak, Maria Consuelo Mura

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The objective of this study was to investigate two single nucleotide polymorphisms located in exon 2 (g.939A > G) and intron 3 (g.4349A > G) of fatty acid binding protein 3 (FABP3) gene in two Iranian sheep breeds, Lori-Bakhtiari and Zel, using polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) approach. The association of the polymorphisms with growth traits and blood parameters was also examined. Results revealed a g.939A > G SNP (single nucleotide polymorphism) in the exon 2 exhibiting three genotypes: AA, AG, and GG. Statistical analysis indicated that this polymorphism significantly influenced blood triglyceride (P < 0.05) and cholesterol (P < 0.08) levels as well as weaning weight (P < 0.05). Animals with AG genotype had the highest blood triglyceride level and weaning weight while the highest amount of blood cholesterol was observed in animals with GG genotype. On the other hand, no significant effect was observed on birth and fat-tail weight traits. The intron 3 (g.4349A > G) was monomorphic across the studied samples. Lori-Bakhtiari breed showed significantly higher blood triglyceride and cholesterol levels, as also birth and weaning weight compared to Zel breed (P < 0.01). Considering that the literature is bereft of any report on the association study between FABP3 SNPs and sheep growth traits and blood parameters, our findings suggest that the investigated polymorphism might be one of the main genetic factors affecting growth and physiological traits in sheep.

Keywords: FABP3 gene, fatness, weaning weight, blood triglyceride, cholesterol, Zel, Lori-Bakhtiari

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Authors: Shabnam Abdi, Behzad Toosi

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Background: Melanoma is the most lethal type of malignant skin cancer in humans and dogs since it spreads rapidly throughout the body. Despite significant advances in treatment, cancer at an advanced stage has a poor prognosis. Hence, more effective treatments are needed to enhance outcomes with fewer side effects. Erythropoietin-producing hepatocellular receptors are the largest family of receptor tyrosine kinases and are divided into two subfamilies, EphA and EphB, both of which play a significant role in disease, especially cancer. Due to their association with proliferation and invasion in many aggressive types of cancer, Eph receptor tyrosine kinases (Eph RTKs) are promising cancer therapy molecules. Because these receptors have not been studied in canine melanoma, we investigated how EphA2 influences survival and tumorigenicity of melanoma cells. Methods: Expression of EphA2 protein in canine melanoma cell lines and human melanoma cell line was evaluated by Western blot. Melanoma cells were transduced with lentiviral particles encoding Eph-targeting shRNAs or non-silencing shRNAs (control) for silencing the expression of EphA2 receptor, and silencing was confirmed by Western blotting and immunofluorescence. The effect of siRNA treatment on cellular proliferation, colony formation, tumorsphere assay, invasion was analyzed by Resazurin assay Matrigel invasion assay, respectively. Results: Expression of EphA2 was detected in canine and human melanoma cell lines. Moreover, stably silencing EphA2 by specific shRNAs significantly and consistently decreased the expression of EphA2 protein in both human and canine melanoma cells. Proliferation, colony formation, tumorsphere and invasion of melanoma cells were significantly decreased in EphA2 siRNA-treated cells compared to control. Conclusion: Our data provide the first functional evidence that the EphA2 receptor plays a critical role in the malignant cellular behavior of melanoma in both human and dogs.

Keywords: ephA2, targeting, melanoma, human, canine

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Authors: Amr A. El Hanafy, Muhammad I. Qureshi, Jamal Sabir, Mohamed Mutawakil, Mohamed M. Ahmed, Hassan El Ashmaoui, Hassan Ramadan, Mohamed Abou-Alsoud, Mahmoud Abdel Sadek

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β-Lactoglobulin (β-LG) is the dominant non-casein whey protein found in bovine milk and of most ruminants. The amino acid sequence of β-LG along with its 3-dimensional structure illustrates linkage with the lipocalin superfamily. Preliminary studies in goats indicated that milk yield can be influenced by polymorphism in genes coding for whey proteins. The aim of this study is to identify and evaluate the incidence of functional polymorphisms in the exonic and intronic portions of β-LG gene in native Saudi goat breeds (Ardi, Habsi, and Harri). Blood samples were collected from 300 animals (100 for each breed) and genomic DNA was extracted using QIAamp DNA extraction Kit. A fragment of the β-LG gene from exon 7 to 3’ flanking region was amplified with pairs of specific primers. Subsequent digestion with Sac II restriction endonuclease revealed two alleles (A and B) and three different banding patterns or genotypes i.e. AA, AB and BB. The statistical analysis showed that β-LG AA genotype had higher milk yield than β-LG AB and β-LG BB genotypes. Nucleotide sequencing of the selected β-LG fragments was done and submitted to GenBank NCBI (Accession No. KJ544248, KJ588275, KJ588276, KJ783455, KJ783456 and KJ874959). Two already established SNPs in exon 7 (+4601 and +4603) and one fresh SNP in the 3’ UTR region were detected in the β-LG fragments with designated AA genotype. The polymorphisms in exon 7 did not produce any amino acid change. Phylogenetic analysis on the basis of nucleotide sequences of native Saudi goats indicated evolutional similarity with the GenBank reference sequences of goat, Bubalus bubalis and Bos taurus.

Keywords: β-Lactoglobulin, Saudi goats, PCR-RFLP, functional polymorphism, nucleotide sequencing, phylogenetic analysis

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Authors: Aoxue Yang, Weili Tian, Haikun Liu

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Brain tumor stem cells (BTSCs) are resistant to therapy and give rise to recurrent tumors. These rare and elusive cells are likely to disseminate during cancer progression, and some may enter dormancy, remaining viable but not increasing. The identification of dormant BTSCs is thus necessary to design effective therapies for glioblastoma (GBM) patients. Glucocorticoids (GCs) are used to treat GBM-associated edema. However, glucocorticoids participate in the physiological response to psychosocial stress, linked to poor cancer prognosis. This raises concern that glucocorticoids affect the tumor and BTSCs. Identifying markers specifically expressed by brain tumor stem cells (BTSCs) may enable specific therapies that spare their regular tissue-resident counterparts. By ribosome profiling analysis, we have identified that glycerol-3-phosphate dehydrogenase 1 (GPD1) is expressed by dormant BTSCs but not by NSCs. Through different stress-induced experiments in vitro, we found that only dexamethasone (DEXA) can significantly increase the expression of GPD1 in NSCs. Adversely, mifepristone (MIFE) which is classified as glucocorticoid receptors antagonists, could decrease GPD1 protein level and weaken the proliferation and stemness in BTSCs. Furthermore, DEXA can induce GPD1 expression in tumor-bearing mice brains and shorten animal survival, whereas MIFE has a distinct adverse effect that prolonged mice lifespan. Knocking out GR in NSC can block the upregulation of GPD1 inducing by DEXA, and we find the specific sequences on GPD1 promotor combined with GR, thus improving the efficiency of GPD1 transcription from CHIP-Seq. Moreover, GR and GPD1 are highly co-stained on GBM sections obtained from patients and mice. All these findings confirmed that GR could regulate GPD1 and loss of GPD1 Impairs Multiple Pathways Important for BTSCs Maintenance GPD1 is also a critical enzyme regulating glycolysis and lipid synthesis. We observed that DEXA and MIFE could change the metabolic profiles of BTSCs by regulating GPD1 to shift the transition of cell dormancy. Our transcriptome and lipidomics analysis demonstrated that cell cycle signaling and phosphoglycerides synthesis pathways contributed a lot to the inhibition of GPD1 caused by MIFE. In conclusion, our findings raise concern that treatment of GBM with GCs may compromise the efficacy of chemotherapy and contribute to BTSC dormancy. Inhibition of GR can dramatically reduce GPD1 and extend the survival duration of GBM-bearing mice. The molecular link between GPD1 and GR may give us an attractive therapeutic target for glioblastoma.

Keywords: cancer stem cell, dormancy, glioblastoma, glycerol-3-phosphate dehydrogenase 1, glucocorticoid receptor, dexamethasone, RNA-sequencing, phosphoglycerides

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Authors: Christopher Benedikt Jakob

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Human beings are intrigued by luxury watches for decades. It is fascinating that customers pay an enormous amount of money for specific wristwatch models. It is fascinating that customers of the luxury watch industry accept a yearly price increase. This behavior increases their desirability even more. Luxury watches are perceived as status symbols, but they are additionally accepted as a currency without the disadvantage of currency fluctuations. It is obvious that the symbolic value is more important as the functional value with reference to the buying-reasons as regards luxury watches. Nowadays human beings do not need a wristwatch to read the time. Tablets, notebooks, smartphones, the watch in the car and watches on public places are used to inform people about the current time. This is one of the reasons why there is a trend that people do not wear wristwatches anymore. Due to these facts, this study has the intention to give answers to the question why people invest an enormous amount of money on the consumption of luxury watches and why those watches are seen as a status symbol. The study examines why the luxury watch industry records significant growth rates. The self-expression model is used as an appropriate methodology to find reasons why human beings purchase specific luxury watches. This evaluative approach further discusses if human beings are aware of their current self and their ideal self and how they express them. Furthermore, the research critically evaluates the people’s social self and their ideal social self. One of the goals is to identify if customers know why they like specific luxury watches and dislike others although they have the same quality and cost comparable prices.

Keywords: luxury watch, brand awareness, buying-behaviour, consumer, self-expression

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Authors: Gyaltsen Dakpa, K. J. Senthil Kumar, Nai-Wen Tsao, Sheng-Yang Wang

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Context: Coronavirus disease 2019 (COVID-19) is a severe respiratory illness caused by the SARS-CoV-2 virus. One of the hallmarks of COVID-19 is a change in metabolism, which can lead to increased severity and mortality. The mechanism of SARS-CoV-2-mediated perturbations of metabolic pathways has yet to be fully understood. Research Aim: This study aimed to investigate the metabolic alteration caused by SARS-CoV-2 spike protein in Phorbol 12-myristate 13-acetate (PMA)-induced human monocytes (THP-1) and to examine the regulatory effect of natural compounds like Antcins A on SARS-CoV-2 spike protein-induced metabolic alteration. Methodology: The study used a combination of proton nuclear magnetic resonance (1H-NMR) and MetaboAnalyst 5.0 software. THP-1 cells were treated with SARS-CoV-2 spike protein or control, and the metabolomic profiles of the cells were compared. Antcin A was also added to the cells to assess its regulatory effect on SARS-CoV-2 spike protein-induced metabolic alteration. Findings: The study results showed that treatment with SARS-CoV-2 spike protein significantly altered the metabolomic profiles of THP-1 cells. Eight metabolites, including glycerol-phosphocholine, glycine, canadine, sarcosine, phosphoenolpyruvic acid, glutamine, glutamate, and N, N-dimethylglycine, were significantly different between control and spike-protein treatment groups. Antcin A significantly reversed the changes in these metabolites. In addition, treatment with antacid A significantly inhibited SARS-CoV-2 spike protein-mediated up-regulation of TLR-4 and ACE2 receptors. Theoretical Importance The findings of this study suggest that SARS-CoV-2 spike protein can cause significant metabolic alterations in THP-1 cells. Antcin A, a natural compound, has the potential to reverse these metabolic alterations and may be a potential candidate for developing preventive or therapeutic agents for COVID-19. Data Collection: The data for this study was collected from THP-1 cells that were treated with SARS-CoV-2 spike protein or a control. The metabolomic profiles of the cells were then compared using 1H-NMR and MetaboAnalyst 5.0 software. Analysis Procedures: The metabolomic profiles of the THP-1 cells were analyzed using 1H-NMR and MetaboAnalyst 5.0 software. The software was used to identify and quantify the cells' metabolites and compare the control and spike-protein treatment groups. Questions Addressed: The question addressed by this study was whether SARS-CoV-2 spike protein could cause metabolic alterations in THP-1 cells and whether Antcin A can reverse these alterations. Conclusion: The findings of this study suggest that SARS-CoV-2 spike protein can cause significant metabolic alterations in THP-1 cells. Antcin A, a natural compound, has the potential to reverse these metabolic alterations and may be a potential candidate for developing preventive or therapeutic agents for COVID-19.

Keywords: SARS-CoV-2-spike, ¹H-NMR, metabolomics, antcin-A, taiwanofungus camphoratus

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Authors: Hyun Ji Hyun, Hyo Sun Suh, Min Kook Kim, Yong Chan Kwon, Byung-Mu Lee

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Scope: This study is focused on the effect of myristic acid on LPS-induced inflammation in RAW 264.7 macrophage cells. Methods and results: For the experiment, RAW 264.7 mouse macrophage cell line was used. Results showed that treatment with myristic acid can attenuate LPS-induced inflammation. Moreover, myristic acid significantly suppressed expression of inflammatory mediators and down-regulating UVB-induced intracellular ROS generation. Furthermore, myristic acid reduced the expression of NF-κB by inhibiting degradation of IκB-α and ERK, JNK, and p38 pathways by inhibiting phosphorylation in RAW 264.7 macrophage cells. Conclusion: Overall, these data suggest that the myristic acid could reduce LPS-induced inflammation. Acknowledgment: This research was supported by the Ministry of Trade, Industry & Energy(MOTIE), Korea Institute for Advancement of Technology(KIAT) through the Encouragement Program for The Industries of Economic Cooperation Region

Keywords: anti-inflammation, myristic acid, ROS, ultraviolet light

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Authors: Mazo Kone, Rachida Salah, Harir Noria

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Background and objectives: c-Cbl and Cbl-b are two members of the Cbl family proteins, with a crucial role of downregulation of tyrosine kinase receptors. They act as E3 ubiquitin ligases and are multivalent adaptor proteins, making them important in maintaining homeostasis in the body. This study investigated the expression level in thymus and testis in normal conditions. Methods: The expression level was assessed by immunochemistry of tissue microarrays of normal thymus and testis biopsies. Results: Cbl-b and c-Cbl proteins were found to be highly expressed in normal testis and thymus, indicated as yellowish brown granules in the cytomembrane and cytoplasm compared to controls. The c-Cbl appears to be more highly expressed than the Cbl-b in the thymus, while c-Cbl appears slightly stronger than Cbl-b in the testis. The thymus was found with a higher grade compared to the testis. Conclusion: In this work we concluded, that in normal condition, thymus tissue expresses more Cbl family proteins(c-Cbl and Cbl-b) than the testis tissue in humans.

Keywords: Human C-Cbl proteins, Human Cbl-b protein, Testis, Thymus

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Authors: Anna Perrone, Federico Martinelli

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Olive fruit is well-known for the high content in secondary metabolites with high interest at nutritional, nutraceutical, antioxidant, and healthy levels. The content of secondary metabolites in olive at harvest may be affected by different water regimes, with significant effects on olive oil composition and quality and, consequently, on its healthy and nutritional features. In this work, a summary of several research studies dealing with the biosynthesis of healthy and nutraceutical metabolites of the secondary metabolism in olive fruit will be reported. The phytochemical findings have been correlated with the expression of key genes involved in polyphenol, terpenoid, and carotenoid biosynthesis and metabolism in response to different development stages and water regimes. Flavonoids were highest in immature fruits, while anthocyanins increased at ripening. In epicarp tissue, this was clearly associated with an up-regulation of the UFGT gene. Olive fruits cultivated under different water regimes were analyzed by metabolomics. This method identified several hundred metabolites in the ripe mesocarp. Among them, 46 were differentially accumulated in the comparison between rain-fed and irrigated conditions. Well-known healthy metabolites were more abundant at a higher level of water regimes. Increased content of polyphenols was observed in the rain-fed fruit; particularly, anthocyanin concentration was higher at ripening. Several secondary metabolites were differentially accumulated between different irrigation conditions. These results showed that these metabolic approaches could be efficiently used to determine the effects of agronomic treatments on olive fruit physiology and, consequently, on nutritional and healthy properties of the obtained extra-virgin olive oil.

Keywords: olea europea, anthocyanins, polyphenols, water regimes

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Authors: Nusrat Masood, Vijaya Dubey, Suaib Luqman

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Caspase 3, a member of cysteine-aspartic acid protease family, is an imperative indicator for cell death particularly when substantiating apoptosis. Thus, caspase 3 is an interesting target for the discovery and development of anticancer agent. We adopted a four level assessment of both terpenoids and flavonoids and thus experimentally performed the enzymatic assay in cell free system as well as in cancer cell line which was validated through real time expression and molecular interaction studies. A significant difference was observed with both the class of natural products indicating terpenoids as better activators of caspase 3 compared to flavonoids both in the cell free system as well as in cell lines. The expression analysis, activation constant and binding energy also correlate well with the enzyme activity. Overall, terpenoids had an unswerving effect on caspase 3 in all the tested system while flavonoids indirectly affect enzyme activity.

Keywords: Caspase 3, terpenoids, flavonoids, activation constant, binding energy

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Authors: Ghulam Muhammad Ali

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Unveiling of genes involved in drought and root architecture using transcriptomic analyses remained fragmented for further improvement of wheat through genome editing. The purpose of this research endeavor was to unveil the variations in different genes implicated in drought tolerance and root architecture in wheat through RNA-seq data analysis. In this study seedlings of 8 days old, 6 cultivars of wheat namely, Batis, Blue Silver, Local White, UZ888, Chakwal 50 and Synthetic wheat S22 were subjected to transcriptomic analysis for root and shoot genes. Total of 12 RNA samples was sequenced by Illumina. Using updated wheat transcripts from Ensembl and IWGC references with 54,175 gene models, we found that 49,621 out of 54,175 (91.5%) genes are expressed at an RPKM of 0.1 or more (in at least 1 sample). The number of genes expressed was higher in Local White than Batis. Differentially expressed genes (DEG) were higher in Chakwal 50. Expression-based clustering indicated conserved function of DRO1and RPK1 between Arabidopsis and wheat. Dendrogram showed that Local White is sister to Chakwal 50 while Batis is closely related to Blue Silver. This study flaunts transcriptomic sequence variations in different cultivars that showed mutations in genes associated with drought that may directly contribute to drought tolerance. DRO1 and RPK1 genes were fetched/isolated for genome editing. These genes are being edited in wheat through CRISPR-Cas9 for yield enhancement.

Keywords: transcriptomic, wheat, genome editing, drought, CRISPR-Cas9, yield enhancement

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Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou

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Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.

Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate

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Authors: Nafiseh Noormohammadi, Ahmad Sobhani Najafabadi

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Hypericins (hypericin and pseudohypericin) are natural napthodianthrone derivatives produced by Hypericum perforatum (St. John’s Wort), which have many medicinal properties such as antitumor, antineoplastic, antiviral, and antidepressant activities. Production and accumulation of hypericin in the plant are influenced by both genetic and environmental conditions. Despite the existence of different high-throughput data on the plant, genetic dimensions of hypericin biosynthesis have not yet been completely understood. In this research, 21 high-quality RNA-seq data on different parts of the plant were integrated into metabolic data to reconstruct a coexpression network. Results showed that a cluster of 30 transcripts was correlated with total hypericin. The identified transcripts were divided into three main groups based on their functions, including hypericin biosynthesis genes, transporters, detoxification genes, and transcription factors (TFs). In the biosynthetic group, different isoforms of polyketide synthase (PKSs) and phenolic oxidative coupling proteins (POCPs) were identified. Phylogenetic analysis of protein sequences integrated into gene expression analysis showed that some of the POCPs seem to be very important in the biosynthetic pathway of hypericin. In the TFs group, six TFs were correlated with total hypericin. qPCR analysis of these six TFs confirmed that three of them were highly correlated. The identified genes in this research are a rich resource for further studies on the molecular breeding of H. perforatum in order to obtain varieties with high hypericin production.

Keywords: hypericin, St. John’s Wort, data mining, transcription factors, secondary metabolites

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Authors: Atul C. Mahajan, A. K. Chakravarty, V. Jamuna, C. S. Patil, Neeraj Kashyap, Bharti Deshmukh, Vijay Kumar

Abstract:

The selection of Sahiwal bulls on the basis of dams best lactation milk yield under breeding programme in herd of the country neglecting fertility traits leads to deterioration in their performances and economy. The goal of this study was to explore polymorphism of CRISP2 gene and their association with semen traits (Post Thaw Motility, Hypo-osmotic Swelling Test, Acrosome Integrity, DNA Fragmentation and capacitation status), scrotal circumference, expected predicted difference (EPD) for milk yield and fertility. Sahiwal bulls included in present study were 60 bulls used in breeding programme as well as 50 young bulls yet to be included in breeding programme. All the Sahiwal bulls were found to be polymorphic for CRISP2 gene (AA, AG and GG) present within exon 7 to the position 589 of CRISP2 mRNA by using PCR-SSCP and Sequencing. Semen analysis were done on 60 breeding bulls frozen semen doses pertaining to four season (winter, summer, rainy and autumn). The scrotal circumference was measured from existing Sahiwal breeding bulls in the herd (n=47). The effect of non-genetic factors on reproduction traits were studied by least-squares technique and the significant difference of means between subclasses of season, period, parity and age group were tested. The data were adjusted for the significant non-genetic factors to remove the differential environmental effects. The adjusted data were used to generate traits like Waiting Period (WP), Pregnancy Rate (PR), Expected Predicted Difference (EPD) of fertility, respectively. Genetic and phenotypic parameters of reproduction traits were estimated. The overall least-squares means of Age at First Calving (AFC), Service Period (SP) and WP were estimated as 36.69 ± 0.18 months, 120.47 ± 8.98 days and 79.78 ± 3.09 days respectively. Season and period of birth had significant effect (p < 0.01) on AFC. AFC was highest during autumn season of birth followed by summer, winter and rainy. Season and period of calving had significant effect (p < 0.01) on SP and WP of sahiwal cows. The WP for Sahiwal cows was standardized based on four developed predicted model for pregnancy rate 42, 63, 84 and 105 days using all lactation records. The WP for Sahiwal cows were standardized as 42 days. A selection criterion was developed for Sahiwal breeding bulls and young Sahiwal bulls on the basis of EPD of fertility. The genotype has significant effect on expected predicted difference of fertility and some semen parameters like post thaw motility and HOST. AA Genotype of CRISP2 gene revealed better EPD for fertility than EPD of milk yield. AA genotype of CRISP2 gene has higher scrotal circumference than other genotype. For young Sahiwal bulls only AA genotypes were present with similar patterns. So on the basis of association of genotype with seminal traits, EPD of milk yield and EPD for fertility status, AA and AG genotype of CRISP2 gene was better for higher fertility in Sahiwal bulls.

Keywords: expected predicted difference, fertility, sahiwal, waiting period

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Authors: Zhang Lei, Zhao Qingrong, Wang Chen, Zhang Sufang, Zhang Hanguo

Abstract:

Larch is the main afforestation and timber tree species in Northeast China, and drought is one of the main factors limiting the growth of Larch and other organisms in Northeast China. In order to further explore the mechanism of Larch drought resistance, PEG was used to simulate drought stress. The full-length sequencing of Larch embryogenic callus under PEG simulated drought stress was carried out by combining Illumina-Hiseq and SMRT-seq. A total of 20.3Gb clean reads and 786492 CCS reads were obtained from the second and third generation sequencing. The de-redundant transcript sequences were predicted by lncRNA, 2083 lncRNAs were obtained, and the target genes were predicted, and a total of 2712 target genes were obtained. The de-redundant transcripts were further screened, and 1654 differentially expressed genes (DEGs )were obtained. Among them, different DEGs respond to drought stress in different ways, such as oxidation-reduction process, starch and sucrose metabolism, plant hormone pathway, carbon metabolism, lignin catabolic/biosynthetic process and so on. This study provides basic full-length sequencing data for the study of Larch drought resistance, and excavates a large number of DEGs in response to drought stress, which helps us to further understand the function of Larch drought resistance genes and provides a reference for in-depth analysis of the molecular mechanism of Larch drought resistance.

Keywords: larch, drought stress, full-length transcriptome sequencing, differentially expressed genes

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Authors: Cengiz Demir, Recep Keşli, Gülşah Aşık

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Objective: This study was carried out with the purpose of defining by using the E test method and determining the antibiotic resistance profiles of Gram-negative anaerobic bacilli isolated from various clinical specimens obtained from patients with suspected anaerobic infections and referred to Medical Microbiology Laboratory of Afyon Kocatepe University, ANS Application and Research Hospital. Methods: Two hundred and seventy eight clinical specimens were examined for isolation of the anaerobic bacteria in Medical Microbiology Laboratory between the 1st November 2014 and 30th October 2015. Specimens were cultivated by using Scheadler agar that 5% defibrinated sheep blood added, and Scheadler broth. The isolated anaerobic Gram-negative bacilli were identified conventional methods and Vitek 2 (ANC ID Card, bioMerieux, France) cards. Antibiotic resistance rates against to penicillin G, clindamycin, cefoxitin, metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem were determined with E-test method for each isolate. Results: Of the isolated twenty-eight anaerobic gram negative bacilli fourteen were identified as the B. fragilis group, 9 were Prevotella group, and 5 were Fusobacterium group. The highest resistance rate was found against penicillin (78.5%) and resistance rates against clindamycin and cefoxitin were found as 17.8% and 21.4%, respectively. Against to the; metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem, no resistance was found. Conclusion: Since high rate resistance has been detected against to penicillin in the study penicillin should not be preferred in empirical treatment. Cefoxitin can be preferred in empirical treatment; however, carrying out the antibiotic sensitivity testing will be more proper and beneficial. No resistance was observed against carbapenem group antibiotics and metronidazole; so that reason, these antibiotics should be reserved for treatment of infectious caused by resistant strains in the future.

Keywords: anaerobic gram-negative bacilli, anaerobe, antibiotics and resistance profiles, e-test method

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Authors: Edison Bicudo

Abstract:

Cell and gene therapies (CGTs) result from the intense manipulation of cells or the use of techniques such as gene editing. They have been increasingly used to tackle rare diseases or conditions of genetic origin, such as cancer. One might expect such a complex scientific field to be dominated by scientific findings and evidence-based explanations. However, people engaged in scientific argumentation also mobilize a range of cognitive operations of which they are not fully aware, in addition to drawing on widely available oral traditions. This paper analyses how experts discussing the potentialities and challenges of CGTs have recourse to a particular kind of prototypical myth. This sociology study, conducted at the University of Sussex (UK), involved interviews with scientists, regulators, and entrepreneurs involved in the development or governance of CGTs. It was observed that these professionals, when voicing their views, sometimes have recourse to narratives where CGTs appear as promising tools for alleviating or curing diseases. This is said to involve much personal, scientific, and financial sacrifice. In his study of traditional narratives, Hogan identified three prototypes: the romantic narrative, moved by the ideal of romantic union; the heroic narrative, moved by the desire for political power; and the sacrificial narrative, where the ideal is plenty, well-being, and health. It is argued here that discourses around CGTs often involve some narratives – or myths – that have a sacrificial nature. In this sense, the development of innovative therapies is depicted as a huge sacrificial endeavor involving biomedical scientists, biotech and pharma companies, and decision-makers. These sacrificial accounts draw on oral traditions and benefit from an emotional intensification that can be easily achieved in stories of serious diseases and physical suffering. Furthermore, these accounts draw on metaphorical understandings where diseases and vectors of diseases are considered enemies or invaders while therapies are framed as shields or protections. In this way, this paper aims to unravel the cognitive underpinnings of contemporary science – and, more specifically, biomedicine – revealing how myths, prototypes, and metaphors are highly operative even when complex reasoning is at stake. At the same time, this paper demonstrates how such hidden cognitive operations underpin the construction of powerful ideological discourses aimed at defending certain ways of developing, disseminating, and governing technologies and therapies.

Keywords: cell and gene therapies, myths, prototypes, metaphors

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