Search results for: CRAC motif

Authors: Yi-Hui Lai, Chih-Hsiang Yang, Li-Chung Hsu

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The inflammasome is a protein complex that modulates caspase-1 activity, resulting in proteolytic cleavage of proinflammatory cytokines such as IL-1β and IL-18, into their bioactive forms. It has been shown that the inflammasomes play a crucial role in the clearance of pathogenic infection and tissue repair. However, dysregulated inflammasome activation contributes to a wide range of human diseases such as cancers and auto-inflammatory diseases. Yet, regulation of NLRP3 inflammasome activation remains largely unknown. We discovered a novel DEAD box protein, whose biological function has not been reported, not only negatively regulates NLRP3 inflammasome activation by interfering NLRP3 inflammasome assembly and cellular localization but also mitigate pyroptosis upon pathogen evasion. The DEAD-box protein is the first DEAD-box protein gets involved in modulation of the inflammasome activation. In our study, we found that caspase-1 activation and mature IL-1β production were largely enhanced upon LPS challenge in the DEAD box-containing protein- deleted THP-1 macrophages and bone marrow-derived macrophages (BMDMs). In addition, this DEAD box-containing protein migrates from the nucleus to the cytoplasm upon LPS stimulation, which is required for its inhibitory role in NLRP3 inflammasome activation. The DEAD box-containing protein specifically interacted with the LRR motif of NLRP3 via its DEAD domain. Furthermore, due to the crucial role of the NLRP3 LRR domain in the recruitment of NLRP3 to mitochondria and binding to its adaptor ASC, we found that the interaction of NLRP3 and ASC was downregulated in the presence of the DEAD box-containing protein. In addition to the mechanical study, we also found that this DEAD box protein protects host cells from inflammasome-triggered cell death in response to broad-ranging pathogens such as Candida albicans, Streptococcus pneumoniae, etc., involved in nosocomial infections and severe fever shock. Collectively, our results suggest that this novel DEAD box molecule might be a key therapeutic strategy for various infectious diseases.

Keywords: inflammasome, inflammation, innate immunity, pyroptosis

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Authors: Manju Kanu, Subrata Sinha, Surabhi Johari

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Viral proteins of Ebola viruses belong to one of the best studied viruses; however no effective prevention against EBOV has been developed. Epitope-based vaccines provide a new strategy for prophylactic and therapeutic application of pathogen-specific immunity. A critical requirement of this strategy is the identification and selection of T-cell epitopes that act as vaccine targets. This study describes current methodologies for the selection process, with Ebola virus as a model system. Hence great challenge in the field of ebola virus research is to design universal vaccine. A combination of publicly available bioinformatics algorithms and computational tools are used to screen and select antigen sequences as potential T-cell epitopes of supertypes Human Leukocyte Antigen (HLA) alleles. MUSCLE and MOTIF tools were used to find out most conserved peptide sequences of viral proteins. Immunoinformatics tools were used for prediction of immunogenic peptides of viral proteins in zaire strains of Ebola virus. Putative epitopes for viral proteins (VP) were predicted from conserved peptide sequences of VP. Three tools NetCTL 1.2, BIMAS and Syfpeithi were used to predict the Class I putative epitopes while three tools, ProPred, IEDB-SMM-align and NetMHCII 2.2 were used to predict the Class II putative epitopes. B cell epitopes were predicted by BCPREDS 1.0. Immunogenic peptides were identified and selected manually by putative epitopes predicted from online tools individually for both MHC classes. Finally sequences of predicted peptides for both MHC classes were looked for common region which was selected as common immunogenic peptide. The immunogenic peptides were found for viral proteins of Ebola virus: epitopes FLESGAVKY, SSLAKHGEY. These predicted peptides could be promising candidates to be used as target for vaccine design.

Keywords: epitope, b cell, immunogenicity, ebola

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Authors: Zahra Dehghani, Hoda Dehghani, Elham Zarenezhad

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1,2,3-Triazole derivatives are important compounds in medicinal chemistry owing to their wide applications in drug discovery. They can readily associate with biologically targets through the hydrogen bonding and dipole interactions. The 1,2,3-triazole core is a key structural motif in many bioactive compounds, exhibiting a broad spectrum of biological activities, such as antiviral, anticancer, anti-HIV, antibiotic, antibacterial, and antimicrobial. Additionally, they have found significant industrial applications as dyes, agrochemicals, corrosion inhibitors, photo stabilizers, and photographic materials. we disclose the synthesis and characterization of 1-azido-3-(aryl-2-yloxy)propan-2-ol drivatives. The chemistry works well with various ß-azido alcohols involving aryloxy, alkoxy and alkyl residues, and also tolerates a wide spectrum of electron-donating and electron-withdrawing functional groups in both alkyne and azide molecules. Most of ß-azidoalcohols used in these experiments were pre-synthesized by the regioselective ring opening reaction of corresponded epoxides with sodium azide, whereas the majority of terminal alkynes were prepared via SN2-type reaction of propargyl bromide and corresponded nucleophiles. To evaluate the bioactivity of title compounds, the in vitro antifungal activity of all compound was investigated against several pathogenic fungi including Candida albicans, Candida krusei, Aspergillus niger, and Trichophyton rubrum , clotrimazole and fluconazole was used as standard antifungal drugs, also To understand the antibacterial activity of synthesized compounds, they were in vitro screened against E. coli and S. aureus as Gram-negative and Gram-positive bacteria, respectively. The in vitro tests have shown the promising antifungal but marginal antibacterial activity against tested fungi and bacteria.

Keywords: biological activities, antibacterial, antifungal, 1, 2, 3-Triazole

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Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy

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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.

Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide

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Authors: Lynette Sheridan Burns, Benjamin J. Matthews

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The context of post-industrial journalism is one in which the material circumstances of mechanical publication have been displaced by digital technologies, increasing the distance between the orthodoxy of the newsroom and the culture of journalistic writing. Content is, with growing frequency, created for delivery via the internet, publication on web-based ‘platforms’ and consumption on screen media. In this environment, the question is not ‘who is a journalist?’ but ‘what is journalism?’ today. The changes bring into sharp relief new distinctions between journalistic work and journalistic labor, providing a key insight into the current transition between the industrial journalism of the 20^th century, and the post-industrial journalism of the present. In the 20^th century, the work of journalists and journalistic labor went hand-in-hand as most journalists were employees of news organizations, whilst in the 21^st century evidence of a decoupling of ‘acts of journalism’ (work) and journalistic employment (labor) is beginning to appear. This 'decoupling' of the work and labor that underpins journalism practice is far reaching in its implications, not least for institutional structures. Under these conditions we are witnessing the emergence of expanded ‘entrepreneurial’ journalism, based on smaller, more independent and agile - if less stable - enterprise constructs that are a feature of creative industries. Entrepreneurial journalism is realized in a range of organizational forms from social enterprise, through to profit driven start-ups and hybrids of the two. In all instances, however, the primary motif of the organization is an ideological definition of journalism. An example is the Scoop Foundation for Public Interest Journalism in New Zealand, which owns and operates Scoop Publishing Limited, a not for profit company and social enterprise that publishes an independent news site that claims to have over 500,000 monthly users. Our paper demonstrates that this journalistic work meets the ideological definition of journalism; conducted within the creative industries using an innovative organizational structure that offers a new, viable post-industrial future for journalism.

Keywords: creative industries, digital communication, journalism, post industrial

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Authors: Jiangang Shen

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Olfactory dysfunction is a common symptom companied by anxiety- and depressive-like behaviors in depressive patients. Chronic stress triggers hormone responses and inhibits the proliferation and differentiation of neural stem cells (NSCs) in the hippocampus and subventricular zone (SVZ)-olfactory bulb (OB), contributing to depressive behaviors and olfactory dysfunction. However, the cellular signaling molecules to regulate chronic stress mediated olfactory dysfunction are largely unclear. Adaptor proteins containing the pleckstrin homology domain, phosphotyrosine binding domain, and leucine zipper motif (APPLs) are multifunctional adaptor proteins. Herein, we tested the hypothesis that APPL2 could inhibit hippocampal neurogenesis by affecting glucocorticoid receptor (GR) signaling, subsequently contributing to depressive and anxiety behaviors as well as olfactory dysfunctions. The major discoveries are included: (1) APPL2 Tg mice had enhanced GR phosphorylation under basic conditions but had no different plasma corticosterone (CORT) level and GR phosphorylation under stress stimulation. (2) APPL2 Tg mice had impaired hippocampal neurogenesis and revealed depressive and anxiety behaviors. (3) GR antagonist RU486 reversed the impaired hippocampal neurogenesis in the APPL2 Tg mice. (4) APPL2 Tg mice displayed higher GR activity and less capacity for neurogenesis at the olfactory system with lesser olfactory sensitivity than WT mice. (5) APPL2 negatively regulates olfactory functions by switching fate commitments of NSCs in adult olfactory bulbs via interaction with Notch1 signaling. Furthermore, baicalin, a natural medicinal compound, was found to be a promising agent targeting APPL2/GR signaling and promoting adult neurogenesis in APPL2 Tg mice and chronic corticosterone-induced depression mouse models. Behavioral tests revealed that baicalin had antidepressant and olfactory-improving effects. Taken together, APPL2 is a critical therapeutic target for antidepressant treatment.

Keywords: APPL2, hippocampal neurogenesis, depressive behaviors and olfactory dysfunction, stress

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Authors: Ivana Prochazkova

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Cognitive linguistics offers biblical scholarship a specific methodological tool for analysis and interpretation of metaphorical expressions. Its methodology makes it possible to study processes involved in constructing the meaning of individual metaphorical expressions and whole conceptual metaphors; to analyze their function in the text; to follow the semantic development of concepts and conceptual domains, and to trace semantic changes and their motivation. The legal language in the Hebrew canon is extremely specific and formalized. Especially in the preambles to the collections of laws in the Pentateuch, more general considerations of the motif of keeping and breaking the law are encountered. This is also true in the psalms and wisdom literature. Legal theory and the philosophy of law deal with these motifs today. Metaphors play an important role in texts that reflect on more general issues. The purpose of this conference contribution is to write all over the central metaphorical concept, conceptual metaphor ךרד תורה (TORAH/LAW IS A JOURNEY), its function in the Torah and principal trends of the further development in the Prophets and the Writings. The conceptual metaphor תורה ךרד (TORAH/LAW IS A JOURNEY) constitutes a coherent system in conjunction with other metaphors that include e.g., conceptual metaphors נחה תורה (TORAH/LAW LEADS); its variant רעה תורה (TORAH IS A SHEPHERD/GUIDE); מקור תורה (TORAH/LAW IS A FOUNTAIN/A SOURCE OF LIFE). Some conceptual metaphors are well known, and their using are conventional (עשׁר תורה TORAH/LAW IS RICHES, שׂשׂון תורה TORAH/LAW IS DELIGHT, דבשׁ תורה TORAH/LAW IS HONEY, שׁמשׁ תורה TORAH/LAW IS SUN ). But some conceptual metaphors are by its occurrence innovative and unique (e.g., שׁריון תורה TORAH /LAW IS BODY ARMOR, כובע תורה TORAH /LAW IS A HELMET, בגד תורה TORAH/LAW IS A GARMENT, etc.). There will be given examples. Conceptual metaphors will be described by means of some 'metaphorical vehicles,' which are Hebrew expressions in the source domain that are repeatedly used in metaphorical conceptualizations of the target domain(s). Conceptual metaphors will be further described by means of 'generic narrative structures,' which are the particular aspects of a conceptual metaphor that emerge during the metaphorical structuring of concepts. They are the units of the metaphorical vehicles – the Hebrew expressions in the source domain – that structure concepts in much the same way that the conceptual metaphor in the target domain does. And finally, they will be described by means of the network of correspondences that exist between metaphorical vehicles – or generic metaphorical structures – and the Hebrew expressions in the target domain.

Keywords: cognitive theology, conceptual metaphor in the Old Testament, conceptual metaphors of the Torah, conceptual domain of law, righteousness, and justice

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Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

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A huge revolution has emerged in genome engineering by the discovery of CRISPR (clustered regularly interspaced palindromic repeats) and CRISPR-associated system genes (Cas) in bacteria. The function of type II Streptococcus pyogenes (Sp) CRISPR/Cas9 system has been confirmed in various species. Other S. thermophilus (St) CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been also reported for preventing phage infection. The CRISPR1-Cas system interferes by cleaving foreign dsDNA entering the cell in a length-specific and orientation-dependant manner. The S. thermophilus CRISPR3-Cas system also acts by cleaving phage dsDNA genomes at the same specific position inside the targeted protospacer as observed in the CRISPR1-Cas system. It is worth mentioning, for the effective DNA cleavage activity, RNA-guided Cas9 orthologs require their own specific PAM (protospacer adjacent motif) sequences. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of target site for the three orthogonals of Cas9 protein, a well-organized procedure will be required for high-throughput and accurate mining of possible target sites in a large genomic dataset. Consequently, we created a reliable procedure to explore potential gRNA sequences for type I (Streptococcus thermophiles), II (Streptococcus pyogenes), and III (Streptococcus thermophiles) CRISPR/Cas systems. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows: i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. The output of such procedure highlights the power of comparative genome mining for different CRISPR/Cas systems. This could yield a repertoire of Cas9 variants with expanded capabilities of gRNA design, and will pave the way for further advance genome and epigenome engineering.

Keywords: CRISPR/Cas systems, gRNA mining, Streptococcus pyogenes, Streptococcus thermophiles

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Authors: Reshu Saxena, R. K. Tripathi

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HIV-1 transmission and spread involves significant host-virus interaction. Potential targets for prevention of HIV-1 lies at the site of mucosal barriers. Thus a better understanding of how HIV-1 infects target cells at such sites and lead their invasion is required, with prime focus on the host determinants regulating HIV-1 spread. HIV-1 Nef is important for viral infectivity and pathogenicity. It promotes HIV-1 replication, facilitating immune evasion by interacting with various host factors and altering cellular pathways via multiple protein-protein interactions. In this study nef was sequenced from HIV-1 patients, and showed specific mutations revealing sequence variability in nef. To explore the difference in Nef functionality based on sequence variability we have studied the effects of HIV-1 Nef in human SupT1 T cell line and (THP-1) monocyte-macrophage cell lines through proteomics approach. 2D-Gel Electrophoresis in control and Nef-transfected SupT1 cells demonstrated several differentially expressed proteins with significant modulation of alpha-enolase. Through further studies, effects of Nef on alpha-enolase regulation were found to be cell lineage-specific, being stimulatory in macrophages/monocytes, inhibitory in T cells and without effect in HEK-293 cells. Cell migration and invasion studies were employed to determine biological function affected by Nef mediated regulation of alpha-enolase. Cell invasion was enhanced in THP-1 cells but was inhibited in SupT1 cells by wildtype nef. In addition, the modulation of enolase and cell invasion remained unaffected by a unique nef variant. These results indicated that regulation of alpha-enolase expression and invasive property of host cells by Nef is sequence specific, suggesting involvement of a particular motif of Nef. To precisely determine this site, we designed a heptapeptide including the suggested alpha-enolase regulating sequence of nef and a nef mutant with deletion of this site. Macrophages/monocytes being the major cells affected by HIV-1 at mucosal barriers, were particularly investigated by the nef mutant and peptide. Both the nef mutant and heptapeptide led to inhibition of enhanced enolase expression and increased invasiveness in THP-1 cells. Together, these findings suggest a possible mechanism of host invasion by HIV-1 through Nef mediated regulation of alpha-enolase and identifies a potential therapeutic target for HIV-1 entry at mucosal barriers.

Keywords: HIV-1 Nef, nef variants, host-virus interaction, tissue invasion

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Authors: Ratna Sharma, Kaveri Dutta

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Assam (India), the gate way to the Northeast India is mainly known for its exquisite silks, the art and craft. The state has a rich collection of traditional jewellery which is unique and exclusive to the state. These jewelleries hold a special place in the heart of the Assamese women. Similarly handloom industry of Assam is basically silk oriented. Among the wild silk, Eri silk fabric has remained as “the poor man’s silk” but it is closely attached to the assamese society, dress for it's warm quality. In view of the changing market trends, fashion and consumer demands, Silk is emerging as a fashion fabric both in India and abroad. In case of Eri silk fabric it has limited use in clothing and accessories. Hence the restructured and redesigned traditional jewellery motifs of Assam (India) over Eri silk products will have greater potential in reviving the decline of art, generate revenue, self employment towards craftsmen and also recognition of the art. The information incorporated in the paper is primary and the data have been collected by purposive sampling method. This work of art was expressed on Eri silk fabric in the form of traditional hand embroidery as it is closely connected with the era of the individual in history of mankind and reflects the personal expression of an entity. For this study selected traditional motifs of Assamese ornaments was used. Some of the popular traditional Assamese jewellery include earrings with exquisite Lokaparo, Keru, Thuriya, Jangphai, etc. An array of necklaces including Golpata, Satsori, Jon biri, Bena, Gejera, Dhol biri, Doog doogi, Biri Moni, Mukuta Moni, Poalmoni, Silikha Moni and Magardana and diversified rings including Senpata, Horinsakua, Jethinejia, bakharpata and others. Selected two motifs each from necklace, earring and finger ring designs. Selected motifs were further developed into 3 categories- the border, the main motif and all over butta followed by placement of developed patterns on products. Products developed were stoles, scarf’s, purses, brooch pins, skirts for women and ties, handkerchief, jackets for men. The developed products were surveyed by selected respondents. From the present study it can be observed that the embellished traditional jewellery motifs resulted in fresh and colourful pattern on developed Eri silk products. Moreover the motifs which were gradually fading among the community itself showed a very good recognition towards art. The embroidered Eri silk fabric also created a huge change in a positive way among craftsman.

Keywords: Art and craft of Assam, eri silk, hand embroidery, traditional Assamese jewellery motifs

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Authors: Maria Ostroukhova, Zhanneta Zalutskaya, Elena Ermilova

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The alternative oxidase (AOX) mediates cyanide-resistant respiration, which bypasses proton-pumping complexes III and IV of the cytochrome pathway to directly transfer electrons from reduced ubiquinone to molecular oxygen. In Chlamydomonas reinhardtii, AOX is a monomeric protein that is encoded by two genes of discrete subfamilies, AOX1 and AOX2. Although AOX has been proposed to play essential roles in stress tolerance of organisms, the role of subfamily AOX2 is largely unknown. In C. reinhardtii, AOX2 was initially identified as one of constitutively low expressed genes. Like other photosynthetic organisms C. reinhardtii cells frequently experience periods of hypoxia. To examine AOX2 transcriptional regulation and role of AOX2 in hypoxia adaptation, real-time PCR analysis and artificial microRNA method were employed. Two experimental approaches have been used to induce the anoxic conditions: dark-anaerobic and light-anaerobic conditions. C. reinhardtii cells exposed to the oxygen deprivation have shown increased AOX2 mRNA levels. By contrast, AOX1 was not an anoxia-responsive gene. In C. reinhardtii, a subset of genes is regulated by transcription factor CRR1 in anaerobic conditions. Notable, the AOX2 promoter region contains the potential motif for CRR1 binding. Therefore, the role of CRR1 in the control of AOX2 transcription was tested. The CRR1-underexpressing strains, that were generated and characterized in this work, exhibited low levels of AOX2 transcripts under anoxic conditions. However, the transformants still slightly induced AOX2 gene expression in the darkness. These confirmed our suggestions that darkness is a regulatory stimulus for AOX genes in C. reinhardtii. Thus, other factors must contribute to AOX2 promoter activity under dark-anoxic conditions. Moreover, knock-down of CRR1 caused a complete reduction of AOX2 expression under light-anoxic conditions. These results indicate that (1) CRR1 is required for AOX2 expression during hypoxia, and (2) AOX2 gene is regulated by CRR1 together with yet-unknown regulatory factor(s). In addition, the AOX2-underexpressing strains were generated. The analysis of amiRNA-AOX2 strains suggested a role of this alternative oxidase in hypoxia adaptation of the alga. In conclusion, the results reported here show that C. reinhardtii AOX2 gene is stress inducible. CRR1 transcriptional factor is involved in the regulation of the AOX2 gene expression in the absence of oxygen. Moreover, AOX2 but not AOX1 functions under oxygen deprivation. This work was supported by Russian Science Foundation (research grant № 16-14-10004).

Keywords: alternative oxidase 2, artificial microRNA approach, chlamydomonas reinhardtii, hypoxia

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Authors: Norhayati Ayob, Shamsu Mohamad

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This paper aims to provide an initial background of the process of making traditional ceramic pottery, focusing on the materials and the influence of culture heritage. Ceramic pottery is one of the hallmarks of Sabah’s heirloom, not only use as cooking and storage containers but also closely linked with folk cultures and heritage. The Bajau Laut ethnic community of Semporna or better known as the Sea Gypsies, mostly are boat dwellers and work as fishermen in the coast. This ethnic community is famous for their own artistic traditional heirloom, especially the traditional hand-made clay stove called Lapohan. It is found that in the daily life of Bajau Laut community, Lapohan (clay stove) is used to prepare the meal and as a food warmer while they are at the sea. Besides, Lapohan pottery conveys symbolic meaning of natural objects, which portrays the identity, and values of Bajau Laut community. It is acknowledged that the basic process of making potterywares was much the same for people all across the world, nevertheless, it is crucial to consider that different ethnic groups may have their own styles and choices of raw materials. Furthermore, it is still unknown why and how the Bajau Laut ethnic of Semporna get started making their own pottery and to survive until today by heavily depending on the raw materials available in Semporna. In addition, the emergent problem faced by the pottery maker in Sabah is the absence of young successor to continue the heirloom legacy. Therefore, this research aims to explore the traditional pottery making in Sabah, by investigating the background history of Lapohan pottery and to propose the classification of Lapohan based on design and motifs of traditional pottery that will be recognised throughout the study. It is postulated that different techniques and forms of making traditional pottery may produce different types of pottery in terms of surface decoration, shape, and size that portrays different cultures. This study will be conducted at Selakan Island, Semporna, which is the only location that still has Lapohan making. This study is also based on the chronological process of making pottery and taboos of the process of preparing the clay, forming, decoration technique, motif application and firing techniques. The relevant information for the study will be gathered from field study, including observation, in-depth interview and video recording. In-depth interviews will be conducted with several potters and the conversation and pottery making process will be recorded in order to understand the actual process of making Lapohan. The findings hope to provide several types of Lapohan based on different designs and cultures, for example, the one with flat-shape design or has round-shape on the top of clay stove will be labeled with suitable name based on their culture. In conclusion, it is hoped that this study will contribute to conservation for traditional pottery making in Sabah as well as to preserve their culture and heirloom for future generations.

Keywords: Bajau Laut, culture, Lapohan, traditional pottery

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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: Jelena Kovačević Vorgučin, Sibila Petenji Arbutina

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The postmodern age brings a rapid development of technology which inevitably leads to man's need to adapt to modern lifestyle. On the one hand, technological achievements have made human life easier, but there are numerous risks involved. Moreover, man's awareness and perception is changing and adapting unconsciously to the world we live in, while communication in the 21st century is predominantly based on the consumption of images. This paper presents sociological aspects of a community which is confined due to turbulent political-economic circumstances and its impact on the development of media literacy in Serbia. Previous researches led to the conclusion that the media culture is on an extremely low level, and that it can have a strong influence on the general development of the society, starting from the youngest segment of the population. Our aim is to use the conceptual authorial photographs inspired by the obtained research results to emphasize the importance that the impact of visual art has in delivering the message, its role in education and in raising awareness of universal social problems. The paper presents a number of stages involved in the conceptual project which is designed to last over a longer period of time in order to facilitate dissemination of information. First, a survey was carried out in several preschool institutions. This resulted in obtaining the necessary information on the habitual use of the medium of television in children and their carers-parents. The second stage focused on the relationship between the parent and the child in TV consumption. Further, an overview of the visual part of the project was made, which consisted of photographs in various dimensions, ranging from miniature to large dimensions, and following various exhibition principles in both gallery and alternative spaces. This stage of the project placed particular emphasis on the non-standard exhibiting formats and alternative exhibition principles which are increasingly present in all kinds of visual art aimed at achieving a higher level of information noticing and memorizing. The motif on the authorial photographs is children's portraits taken while they are watching different television contents, with emphasis on their emotional response. The importance of the medium of TV is particularly emphasized due to the fact that its consumption is the highest, even though there are newer and more advanced information-technological achievements. The already realized part of the project was used for an analysis of the results in the last stage of the project, which led to the conclusion that the response to the entire visual expression campaign was extremely positive, and action as such very useful indeed. The results obtained speak in favour of widening and continuation of the project, both on a greater number of sites locally as well as in other communities in Serbia with the aim of guiding people towards meaningful consumption of the television medium.

Keywords: alternative space exhibiting, children and TV, conceptual portrait photography, media literacy

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Authors: Kaveri Dutta, Ratna Sharma

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Indian traditional textile is a synthesis of various cultures. Art and crafts of a country showcases the rich cultural and artistic history of that nation. Prehistorically Indian handicrafts were basically made for day to day use; the yearning for aesthetic application soon saw the development of flooding designs and motifs. Similarly, Bagh print a traditional hand block Print with natural colours an Indian handicraft practiced in Bagh, Madhya Pradesh(India). Bagh print has its roots in Sindh, which is now a part of Pakistan. The present form of Bagh printing actually started in 1962 when the craftsmen migrated from Manavar to the neighboring town of Bagh situated in Madhya Pradesh and hence Bagh has always been associated with this printing style. Bagh printing basically involved blocks that are carved onto motifs that represent flora such as Jasmine, Mushroom leheriya and so on. There are some prints that were inspired by the jaali work that embellished the Taj Mahal and various other forts. Inspiration is also drawn from the landscapes and geometrical figures. The motifs evoke various moods in the serenity of the prints and that is the catchy element of Bagh prints. The development in this traditional textile is as essential as in another field. Nowadays fashion trends are fragile and innovative changes over existing fashion field in the short span is the demand of times. We must make efforts to preserve this cultural heritage of arts and crafts and this is done either by documenting the various ancient traditions or by making a blend of it. Since this craft is well known over the world, but the need is to document the original motif, fabric, technology and colors used in contemporary fashion. Hence keeping above points in mind this study on bagh print textiles of Madhya Pradesh work has been formulated. The information incorporated in the paper was based on secondary data taken from relevant books, journals, museum visit and articles. Besides for the demographic details and working profile of the artisans dealt with printing, an interview schedule was carried out in three regions of Madhya Pradesh. This work of art was expressed in Cotton fabric. For this study selected traditional motifs for Bang printing was used. Some of the popular traditional Bagh motifs are Jasmine, Mushroom leheriya, geometrical figures and jaali work. The Bagh printed cotton fabrics were developed into a range of men’s ethic wear in combination with embroideries from Rajasthan. Products developed were bandhgala jackets, kurtas, serwani and dupattas. From the present study, it can be observed that the embellished traditional Bang printed range of ethnic men’s wear resulted in the fresh and colourful pattern. The embroidered Bagh printed cotton fabric also created a huge change in a positive way among artisans of the three regions.

Keywords: art and craft of Madhya Pradesh, evolution of printing in India, history of Bagh printing, sources of inspiration

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Authors: Marcel Verwey, Anna M. Joubert, Elsie M. Nolte, Wolfgang Dohle, Barry V. L. Potter, Anne E. Theron

Abstract:

A tetrahydroisoquinolinone (THIQ) core can be used to mimic the A,B-ring of colchicine site-binding microtubule disruptors such as 2-methoxyestradiol in the design of anti-cancer agents. Steroidomimeric microtubule disruptors were synthesized by introducing C'2 and C'3 of the steroidal A-ring to C'6 and C'7 of the THIQ core and by introducing a decorated hydrogen bond acceptor motif projecting from the steroidal D-ring to N'2. For this in vitro study, four non-steroidal THIQ-based analogues were investigated and comparative studies were done between the non-sulphamoylated compound STX 3450 and the sulphamoylated compounds STX 2895, STX 3329 and STX 3451. The objective of this study was to investigate the modes of cell death induced by these four THIQ-based analogues in A549 lung carcinoma epithelial cells and metastatic breast adenocarcinoma MDA-MB-231 cells. Cytotoxicity studies to determine the half maximal growth inhibitory concentrations were done using spectrophotometric quantification via crystal violet staining and lactate dehydrogenase (LDH) assays. Microtubule integrity and morphologic changes of exposed cells were investigated using polarization-optical transmitted light differential interference contrast microscopy, transmission electron microscopy and confocal microscopy. Flow cytometric quantification was used to determine apoptosis induction and the effect that THIQ-based analogues have on cell cycle progression. Signal transduction pathways were elucidated by quantification of the mitochondrial membrane integrity, cytochrome c release and caspase 3, -6 and -8 activation. Induction of autophagic cell death by the THIQ-based analogues was investigated by morphological assessment of fluorescent monodansylcadaverine (MDC) staining of acidic vacuoles and by quantifying aggresome formation via flow cytometry. Results revealed that these non-steroidal microtubule disrupting analogues inhibited 50% of cell growth at nanomolar concentrations. Immunofluorescence microscopy indicated microtubule depolarization and the resultant mitotic arrest was further confirmed through cell cycle analysis. Apoptosis induction via the intrinsic pathway was observed due to depolarization of the mitochondrial membrane, induction of cytochrome c release as well as, caspase 3 activation. Potential involvement of programmed cell death type II was observed due to the presence of acidic vacuoles and aggresome formation. Necrotic cell death did not contribute significantly, indicated by stable LDH levels. This in vitro study revealed the induction of the intrinsic apoptotic pathway as well as possible involvement of autophagy after exposure to these THIQ-based analogues in both MDA-MB-231- and A549 cells. Further investigation of this series of anticancer drugs still needs to be conducted to elucidate the temporal, mechanistic and functional crosstalk mechanisms between the two observed programmed cell deaths pathways.

Keywords: apoptosis, autophagy, cancer, microtubule disruptor

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Authors: Ram Sharma, Esha Chatterjee, Santosh Kumar Guru, Kunal Nepali

Abstract:

A tractable anti-lung cancer agent was identified via the installation of a Ring C expanded synthetic analogue of the alkaloid vasicinone [7,8,9,10-tetrahydroazepino[2,1-b] quinazolin-12(6H)-one (TAZQ)] as a surface recognition part in the HDAC inhibitory three-component model. Noteworthy to mention that the candidature of TAZQ was deemed suitable for accommodation in HDAC inhibitory pharmacophore as per the results of the fragment recruitment process conducted by our laboratory. TAZQ was pinpointed through the fragment screening program as a synthetically flexible fragment endowed with some moderate cell growth inhibitory activity against the lung cancer cell lines, and it was anticipated that the use of the aforementioned fragment to generate hydroxamic acid functionality (zinc-binding motif) bearing HDAC inhibitors would boost the antitumor efficacy of TAZQ. Consistent with our aim of applying epigenetic targets to the treatment of lung cancer, a strikingly potent anti-lung cancer scaffold (compound 6) was pinpointed through a series of in-vitro experiments. Notably, the compounds manifested a magnificent activity profile against KRAS and EGFR mutant lung cancer cell lines (IC50 = 0.80 - 0.96 µM), and the effects were found to be mediated through preferential HDAC6 inhibition (IC50 = 12.9 nM). In addition to HDAC6 inhibition, the compounds also elicited HDAC1 and HDAC3 inhibitory activity with an IC50 value of 49.9 nM and 68.5 nM, respectively. The HDAC inhibitory ability of compound 6 was also confirmed from the results of the western blot experiment that revealed its potential to decrease the expression levels of HDAC isoforms (HDAC1, HDAC3, and HDAC6). Noteworthy to mention that complete downregulation of the HDAC6 isoform was exerted by compound 6 at 0.5 and 1 µM. Moreover, in another western blot experiment, treatment with hydroxamic acid 6 led to upregulation of H3 acK9 and α-Tubulin acK40 levels, ascertaining its inhibitory activity toward both the class I HDACs and Class II B HDACs. The results of other assays were also encouraging as treatment with compound 6 led to the suppression of the colony formation ability of A549 cells, induction of apoptosis, and increase in autophagic flux. In silico studies led us to rationalize the results of the experimental assay, and some key interactions of compound 6 with the amino acid residues of HDAC isoforms were identified. In light of the impressive activity spectrum of compound 6, a pH-responsive nanocarrier (hyaluronic acid-compound 6 nanoparticles) was prepared. The dialysis bag approach was used for the assessment of the nanoparticles under both normal and acidic circumstances, and the pH-sensitive nature of hyaluronic acid-compound 6 nanoparticles was confirmed. Delightfully, the nanoformulation was devoid of cytotoxicity against the L929 mouse fibroblast cells (normal settings) and exhibited selective cytotoxicity towards the A549 lung cancer cell lines. In a nutshell, compound 6 appears to be a promising adduct, and a detailed investigation of this compound might yield a therapeutic for the treatment of lung cancer.

Keywords: HDAC inhibitors, lung cancer, scaffold, hyaluronic acid, nanoparticles

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Authors: Michelle Maurer, Antonia Last, Mark S. Gresnigt, Bernhard Hube, Alexander S. Mosig

Abstract:

The intestinal epithelium forms an essential barrier to prevent translocation of microorganisms, toxins or other potentially harmful molecules into the bloodstream. In particular, dendritic cells of the intestinal epithelium orchestrate an adapted response of immune tolerance to commensals and immune defense against invading pathogens. Systemic inflammation is typically associated with a dysregulation of this adapted immune response and is accompanied by a disruption of the epithelial and endothelial gut barrier which enables dissemination of pathogens within the human body. To understand the pathophysiological mechanisms underlying the inflammation-associated gut barrier breakdown, it is crucial to elucidate the complex interplay of the host and the intestinal microbiome. A microfluidically perfused three-dimensional intestine-on-chip model was established to emulate these processes in the presence of immune cells, commensal bacteria, and facultative pathogens. Multi-organ tissue flow (MOTiF) biochips made from polystyrene were used for microfluidic perfusion of the intestinal tissue model. The biochips are composed of two chambers separated by a microporous membrane. Each chamber is connected to inlet and outlet channels allowing independent perfusion of the individual channels and application of microfluidic shear stress. Human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages and intestinal epithelial cells (Caco-2) were assembled on the biochip membrane. Following 7 – 14 days of growth in the presence of physiological flow conditions, the epithelium was colonized with the commensal bacterium Lactobacillus rhamnosus, while the endothelium was perfused with peripheral blood mononuclear cells (PBMCs). Additionally, L. rhamnosus was co-cultivated with the opportunistic fungal pathogen Candida albicans. Within one week of perfusion, the epithelial cells formed self-organized and well-polarized villus- and crypt-like structures that resemble essential morphological characteristics of the human intestine. Dendritic cells were differentiated in the epithelial tissue that specifically responds to bacterial lipopolysaccharide (LPS) challenge. LPS is well-tolerated at the luminal epithelial side of the intestinal model without signs of tissue damage or induction of an inflammatory response, even in the presence of circulating PBMC at the endothelial lining. In contrast, LPS stimulation at the endothelial side of the intestinal model triggered the release of pro-inflammatory cytokines such as TNF, IL-1β, IL-6, and IL-8 via activation of macrophages residing in the endothelium. Perfusion of the endothelium with PBMCs led to an enhanced cytokine release. L. rhamnosus colonization of the model was tolerated in the immune competent tissue model and was demonstrated to reduce damage induced by C. albicans infection. A microfluidic intestine-on-chip model was developed to mimic a systemic infection with a dysregulated immune response under physiological conditions. The model facilitates the colonization of commensal bacteria and co-cultivation with facultative pathogenic microorganisms. Both, commensal bacteria alone and facultative pathogens controlled by commensals, are tolerated by the host and contribute to cell signaling. The human intestine-on-chip model represents a promising tool to mimic microphysiological conditions of the human intestine and paves the way for more detailed in vitro studies of host-microbiota interactions under physiologically relevant conditions.

Keywords: host-microbiota interaction, immune tolerance, microfluidics, organ-on-chip

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