Search results for: host preference

Authors: W. Abu Rayyan, A. Singh, A. M. Al-Jaafreh, W. Abu Dayyih, M. Bustami, S. Salem, N. Seder, K. Schröppel

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Hyphal growth and the transcriptional regulation to the host environment are key issues during the pathogenesis of C. albicans. Tec1p is the C. albicans homolog of a TEA transcription factor family, which share a conserved DNA-binding TEA domain in their N-terminal. In order to define a structure-function relationship of the C. albicans Tec1p protein, we constructed several mutations on the N terminal, C terminal or in the TEA binding domain itself by homologous recombination technology. The modifications in the open reading frame of TEC1 were tested for reconstitution of the morphogenetic development of the tec1/tec1 mutant strain CaAS12. Mutation in the TEA consensus sequence did not confer transition to hyphae whereas the reconstitution of the full-length Tec1p has reconstituted hyphal development. A deletion in one of glutamine-rich regions either in the Tec1p N-terminal or the C-terminal in regions of 53-212 or 637–744 aa, respectively, did not restore morphological development in mutant CaAS12 strain. Whereas, the reconstitution with Tec1p mutants other than the glutamate-rich region has restored the morphogenetic switch. Additionally, the deletion of the glutamine-rich region has attenuated the invasive growth and the heat shock resistance of C. albicans. In conclusion, we show that a glutamine-rich region of Tec1p is essential for the hyphal development and mediating adaptation to the host environment of C. albicans.

Keywords: Candida albicans, morphogenetic development, TEA domain, hyphal formation, TEC1

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Authors: Afees Adebayo Salam

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This paper attempts an in-depth analysis of the economic impact of Ogbomoso migrant community in the Jos metropolis. It discusses the factors that motivated a sizeable number of Ogbomoso people (from southwestern Nigeria) to leave their hometown for a new place/space in Jos (northern Nigeria). It examines the historical antecedent of Ogbomoso migrants in northern Nigeria with emphasis on Jos metropolis. The movement of Ogbomoso migrants to Jos was dictated by the economic and social challenges of colonial and post-colonial periods. The political crisis of the 1960s was a contributory factor to the process of Ogbomoso migration to other parts of Nigeria. In the aftermath, many people migrated from Ogbomoso to different parts of the country and beyond to seek for better economic opportunities. The establishment of Ogbomoso migrant community in Jos was dated back to the colonial era when taxation was introduced by the British. Many people could not pay these taxes from their peasant farming activities, while some embarked on migration to places such as Jos, Kaduna, Kano, Keffi and Bauchi due to the harsh economic situation at home. Their settlement in Jos brought about success in several spheres of human endeavours. Ogbomoso migrants dominated both paid jobs and private business sector such as textile merchants, food stuff sellers, herbalists, printers, transporters, and religious missionaries, as well as clerical officers in the government establishments. Their remittances were invested in different sectors of Ogbomoso economy. The migrants had in one way or the other contributed to the socio-economic development of their host community in Jos as entrepreneurs. Branches of such industries were located in their hometown of Ogbomoso as a clear demonstration of community development. The remittance pattern of the migrants has transformed Ogbomoso to enviable position. Moreover, the economic success of Ogbomoso migrants over the period under review indicates the process of nation building due to peaceful nature of inter-ethnic engagements between Ogbomoso migrants and their host community in Jos. Therefore, the paper makes use of oral, archival and secondary sources to analyse the processes of migration and its economic impact. Oral interviews were conducted in Ogbomoso town with veteran migrants and their family members. Interviews were also conducted in Jos with the indigenous host community as well as other urban residents. Archival materials were obtained from Arewa House Archives and the National Archives, Kaduna and the National Archives, Ibadan.

Keywords: Ogbomoso migrants, Jos metropolis, community development, economic impact

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Authors: Silke Bustamante, Andrea Pelzeter, Andreas Deckmann, Rudi Ehlscheidt, Franziska Freudenberger

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Against the background of demographic changes and the need for motivated and talented employees, companies have a growing interest in being perceived as an attractive employer and gain a competitive edge in the market for talents. Research indicates that corporate social responsibility plays an important role employer choice. This is assumed to be particularly true for the cohorts of the so-called generations y younger applicants born later. Several studies identified cultural, educational and generational differences in CSR preferences. However, until now literature that links preferences for employers’ responsibility with the individual value orientation of the job applicant is scarce. This paper seeks to close this gap and analyzes the preference structure of 500 German university graduates for CSR-related and non-CSR-related employer attributes contingent on their value orientation. In a first step, we identified 17 CSR related attributes in five CSR dimensions (ecology, community, workplace, governance, and market) as well as 11 traditional attributes such as salary, reputation, personal development etc. based on literature review and focus groups. We then used an adapted Conjoint Analysis framework in order to understand the relative preference of each attribute for each participant. Additionally, we surveyed the value orientation of participants based on the European Social Survey (ESS) 21-item human values scale that allow a differentiation of participants into clusters of value orientation. The results allow us not only to operationalize preferences for CSR and other important employer attributes, but – more importantly – to answer the question how different values structures play together with CSR preferences of potential employees. They hereby allow companies to customize employee directed messages to their respective target group and design their employer brand accordingly.

Keywords: corporate social responsibility, employer attractiveness, employer brand, recruiting, values

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Authors: Excellent Hansda

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Over the past years, a lot of research has been on the ethnic enclaves from historical, sociological and economic point of view. However there exist a research gap in the built environment and spatial layout of these areas. When immigrants (People) assimilate in a different place, they struggle to preserve their original identity to maintain their heritage. Then there is the Place, which is the physical manifestation of the heritage, shown through streetscape and architecture. Together 'People and Place' form a relationship with the authenticity of the enclave. As immigrants come in the host country, they try to bring their culture into the place, but at the same time, the culture of the host country also affects the immigrants. This creates conflicts not only in the lifestyle and culture of the immigrants, but also the built characteristics of the place. In the midst of such conflicts, one may easily question the authenticity of an ethnic enclave. In Malaysia, a number of ethnic enclaves emerged due to trade during the medieval times. Little India is one among the other ethnic enclaves present in Chulia Street in Malaysia. The study investigates the factors of 'Place and People', affecting the authenticity of a little India, in the context of an evolving state of Penang in Malaysia. The study is carried through extensive literature review of existing data, followed by observations drawn by visual analysis, discussions and interviews with the stakeholders of the study area. The findings of this research suggest the contribution of 'people and places' in the process of place making in an ethnic enclave. The findings are essential for conservation and further development of ethnic enclaves.

Keywords: conservation, ethnic enclaves, heritage, identity

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Authors: Hanaa M. Abu El Einin, Ahmed T. Sharaf El- Din

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Biomphalaria snails play an integral role in the transmission of Schistosoma mansoni, the causative agent for human schistosomiasis. Two species of Biomphalaria were reported from Egypt, Biomphalaria alexandrina and Biomphalaria glabrata, and later on a hybrid of B. alexandrina and B. glabrata was reported in streams at Nile Delta. All were known to be excellent hosts of S. mansoni. Host-parasite relationship can be viewed in terms of snail susceptibility and parasite infectivity. The objective of this study will highlight the progress that has been made in using molecular approaches to describe the correct identification of snail species that participating in transmission of schistosomiasis, rapid diagnose of infection in addition to susceptibility and resistance type. Snails were identified using of molecular methods involving Randomly Amplified Polymorphic DNA (RAPD), Polymerase Chain Reaction, Restriction Fragment Length Polymorphisms (PCR-RFLP) and Species - specific- PCR. Molecular approaches to diagnose parasite in snails from Egypt: Nested PCR assay and small subunit (SSU) rRNA gene. Also RAPD PCR for study susceptible and resistance phenotype. The results showed that RAPD- PCR, PCR-RFLP and species-specific-PCR techniques were confirmed that: no evidence for the presence of B. glabrata in Egypt, All Biomphalaria snails collected identified as B. alexandrina snail i-e B alexandrinia is a common and no evidence for hybridization with B. glabrata. The adopted specific nested PCR assay revealed much higher sensitivity which enables the detection of S. mansoni infected snails down to 3 days post infection. Nested PCR method for detection of infected snails using S. mansoni fructose -1,6- bisphosphate aldolase (SMALDO) primer, these primers are specific only for S. mansoni and not cross reactive with other schistosomes or molluscan aldolases Nested PCR for such gene is sensitive enough to detect one cercariae. Genetic variations between B. alexandrina strains that are susceptible and resistant to Schistosoma infec¬tion using a RAPD-PCR showed that 39.8% of the examined snails collected from the field were resistant, while 60.2% of these snails showed high infection rates. In conclusion the genetics of the intermediate host plays a more important role in the epidemiological control of schistosomiasis.

Keywords: biomphalaria, molecular differentiation, parasite detection, schistosomiasis

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Authors: Azaz Zaman, Mirazur Rahman

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Capital market is a crucial financial market place where companies and the government can raise long-term funds and, at the same time, investors get the opportunity to invest in the listed companies. Capital markets play a vital role not only in shifting the funds from surplus entity to deficit for investment, but also in the overall economic development of any developing country like Bangladesh. Being the first and biggest capital market of Bangladesh, Dhaka Stock Exchange (DSE) is the prime bourse of the country. The differences in the investment preference— among three broad categories of investors in DSE including individual investors, institutional investors, and government— are easily observed. Authors of this article have used five categories of investors such as sponsors or directors of the company, institutional investors, foreign investors, government, and the general public in order to present a comparative analysis of their investment patterns. Obtaining data on the percentage of investment by these five types of investors in different sectors from the DSE website, this study aims to analyze the sector-wise investment preference of these investors using August 2018 data. The study has found that the sponsors or directors of the company have the highest percentage of investment in the textile industry which is close to 16%. The Bangladesh government, as an investor, has the highest percentage of investment in the fuel & power sector, approximately 32%. It has also found that the mutual funds' sector is mostly financed by institutional investors, nearly 28%. Foreign investors have their most investments in the banking sector, which is close to 22%. It has also revealed that the textile sector is mostly financed by the general public, close to 17%. Nevertheless, general public, surprisingly, has the lowest percentage of investment in the telecommunication sector, which is 0.10%.

Keywords: stock market investment, Dhaka stock exchange, capital market, Bangladesh

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Authors: York Neudel

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The phenomenon of tourism has been considered as tool to overcome cultural frontiers, to comprehend the other and to cope with mutual mistrust and suspicion. Well, that has been a myth, at least when it comes to mass-tourism. Other approaches, like community-based tourism, still are based on the idea of embracing the other in order to help or to understand the cultural difference. In 1997, two American NGOs incentivized a tourism-project in a community in the highlands of Ecuador, in order to protect the cloud forest from destructive exploitation of its own inhabitants. Nineteen years after that, I analyze in this investigation the interactions between the Ecuadorian hosts in the mestizo-community of Yunguilla and the foreign tourist in the quest for “authentic life” in the Ecuadorian cloud forest. As a sort of “contemporary pilgrim” the traveller tries to find authenticity in other times and places far away from their everyday life in Europe or North America. Therefore, tourists are guided by stereotypes and expectations that are produced by the touristic industry. The host, on the other hand, has to negotiate this pre-established imaginary. That generates a kind of theatre-play with front- and backstage in organic gardens, little fabrics and even private housing, since this alternative project offers to share the private space of the host with the tourist in the setting the community-based tourism. In order to protect their privacy, the community creates new hybrid spaces that oscillate between front- and backstages that culminates in a game of hide and seek – a phenomenon that promises interesting frictions for an anthropological case-study.

Keywords: Tourism, Authenticity, Community-based tourism, Ecuador, Yunguilla

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Authors: Andrea Hernandez Hurtado

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Research on social networks (SNs) -- the interactions individuals share with others has shed important light in helping to explain differential use of variable linguistic forms, both in L1s and L2s. Nevertheless, the acquisition of nonstandard L2 Spanish in the Region of Murcia, Spain, and how learners interact with other speakers while sojourning there have received little attention. Murcian Spanish (MuSp) was widely influenced by Panocho, a divergent evolution of Hispanic Latin, and differs from the more standard Peninsular Spanish (StSp) in phonology, morphology, and lexicon. For instance, speakers from this area will most likely palatalize diminutive endings, producing animalico [̩a.ni.ma.ˈli.ko] instead of animalito [̩a.ni.ma.ˈli.to] ‘little animal’. Because L1 speakers of the area produce and prefer salient regional lexicon and morphology (particularly the palatalized diminutive -ico) in their speech, the current research focuses on how international residents in the Region of Murcia use Spanish: (1) whether or not they acquire (perceptively and/or productively) any of the salient regional features of MuSp, and (2) how their SNs explain such acquisition. This study triangulates across three tasks -recognition, production, and preference- addressing both lexicon and morphology, with each task specifically created for the investigation of MuSp features. Among other variables, the effects of L1, residence, and identity are considered. As an ongoing dissertation research, data are currently being gathered through an online questionnaire. So far, 7 participants from multiple nationalities have completed the survey, although a minimum of 25 are expected to be included in the coming months. Preliminary results revealed that MuSp lexicon and morphology were successfully recognized by participants (p<.001). In terms of regional lexicon production (10.0%) and preference (47.5%), although participants showed higher percentages of StSp, results showed that international residents become aware of stigmatized lexicon and may incorporate it into their language use. Similarly, palatalized diminutives (production 14.2%, preference 19.0%) were present in their responses. The Social Network Analysis provided information about participants’ relationships with their interactants, as well as among them. Results indicated that, generally, when residents were more immersed in the culture (i.e., had more Murcian alters) they produced and preferred more regional features. This project contributes to the knowledge of language variation acquisition in L2 speakers, focusing on a stigmatized Spanish dialect and exploring how stigmatized varieties may affect L2 development. Results will show how L2 Spanish speakers’ language is affected by their stay in Murcia. This, in turn, will shed light on the role of SNs in language acquisition, the acquisition of understudied and marginalized varieties, and the role of immersion on language acquisition. As the first systematic account on the acquisition of L2 Spanish lexicon and morphology in the Region of Murcia, it lays important groundwork for further research on the connection between SNs and the acquisition of regional variants, applicable to Murcia and beyond.

Keywords: international residents, L2 Spanish, lexicon, morphology, nonstandard language acquisition, social networks

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Authors: Umer Ayyaz Aslam Sheikh

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Non Apis bee like the bumblebees are important due to their utilization of diverse floral plants and belong to the richest and most conspicuous flower visitors in alpine, temperate and arctic environments for pollination in both natural and managed cropping systems. These bees generally construct underground nests and habitat devastation and crumbling are major causes for their decline in nature. The present study was conducted in the Himalayan range of Azad Jammu, and Kashmir, Pakistan, surveys were conducted during the early spring season to observe maximum Bombus haemorrhoidalis queens (emerged after winter diapauses) searching for a nesting place. Whole study area was grouped into four types of landscape (open field, relatively open , relatively wooded and wooded), five habitat types (field, field boundary, pasture forest boundary and forest) and these habitat further grouped into four different patch types including withered grass, new grass, tussocks and stones and moss. Maximum nest seeking bumblebee queens preferred relatively open field landscape followed by open fields and forest boundaries. Field boundaries were recorded as most proffered habitat along with withered grasses for nesting sites of B. haemorrhoidalis queens. A wooded landscape with stone and moss type of patches were found least preferred nesting sites. This study will be helpful in the future for conservation program this for declining bumblebee species in this region. It will also provide the baseline for the conservation of other bumblebee species of the world.

Keywords: bumblebee, Bombus haemorrhoidalis, habitat, nest seeking preference, Pakistan

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Authors: Mohammad Shafiul Alam, Mineo Saito

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Graphene-based materials have attracted much attention because they are candidates for post silicon materials. Since controlling of impurities is necessary to achieve nano device, we study hydrogen impurity in multi-layer graphene. We perform local spin Density approximation (LSDA) in which the plane wave basis set and pseudopotential are used. Previously hydrogen monomer and dimer in graphene is well theoretically studied. However, hydrogen on multilayer graphene is still not clear. By using first-principles electronic structure calculations based on the LSDA within the density functional theory method, we studied hydrogen monomers and dimers in two-layer graphene. We found that the monomers are spin-polarized and have magnetic moment 1 µB. We also found that most stable dimer is much more stable than monomer. In the most stable structures of the dimers in two-layer graphene, the two hydrogen atoms are bonded to the host carbon atoms which are nearest-neighbors. In this case two hydrogen atoms are located on the opposite sides. Whereas, when the two hydrogen atoms are bonded to the same sublattice of the host materials, magnetic moments of 2 µB appear in two-layer graphene. We found that when the two hydrogen atoms are bonded to third-nearest-neighbor carbon atoms, the electronic structure is nonmagnetic. We also studied hydrogen monomers and dimers in three-layer graphene. The result is same as that of two-layer graphene. These results are very important in the field of carbon nanomaterials as it is experimentally difficult to show the magnetic state of those materials.

Keywords: first-principles calculations, LSDA, multi-layer gra-phene, nanomaterials

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Authors: Hajer Habib, Ghazi Boulila

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The objective of this paper is to present our contribution to the theoretical literature through a simple theoretical model dealing with the effect of transferring funds on the labor market of the countries of origin and on the other hand to test this relationship empirically in the case of Tunisia. The methodology used consists of estimating a panel of the nine main destinations of the Tunisian diaspora in Europe between 1994 and 2014 in order to better value the net effect of these migratory financial flows on unemployment through population growth. The empirical results show that the main factors explaining the decision to emigrate are the economic factors related mainly to the income differential, the demographic factors related to the differential age structure of the origin and host populations, and the cultural factors linked basically to the mastery of the language. Indeed, the stock of migrants is one of the main determinants of the transfer of migratory funds to Tunisia. But there are other variables that do not lack importance such as the economic conditions linked by the host countries. This shows that Tunisian migrants react more to economic conditions in European countries than in Tunisia. The economic situation of European countries dominates the numbers of emigrants as an explanatory factor for the amount of transfers from Tunisian emigrants to their country of origin. Similarly, it is clear that there is an indirect effect of transfers on unemployment in Tunisia. This suggests that the demographic transition conditions the effects of transferring funds on the level of unemployment.

Keywords: demographic changes, international migration, labor market, remittances

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Authors: Bahareh Mostofian

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Foreign Direct Investment (FDI) has long been recognized as a crucial driver of economic growth and development in less-developed countries and their integration into Global Production Networks (GPNs). FDI not only brings capital from the core countries but also technology, innovation, and know-how knowledge that can upgrade the capabilities of host automotive industries. On the other hand, FDI can also have negative impacts on host countries if it leads to significant import dependency. In the case of the Iranian automotive sector, the industry greatly benefited from FDI, with Western carmakers dominating the market. Over time, various types of know-how knowledge, including joint ventures (JVs), trade licenses, and technical assistance, have been provided, helping Iran upgrade its automotive industry. While after the severe geopolitical obstacles imposed by both the EU and the U.S., the industry became over-reliant on the car and spare parts imports, and the lack of emphasis on knowledge transfer further affected the growth and development of the Iranian automotive sector. To address these challenges, current research has adopted a descriptive-analytical methodology to illustrate the gradual changes accrued with foreign suppliers through FDI. The research finding shows that after the two-phase imposed sanctions, the detrimental linkages created by overreliance on the car and spare parts imports without any industrial upgrading negatively affected the growth and development of the national and assembled products of the Iranian automotive sector.

Keywords: less-developed country, FDI, GPNs, automotive industry, Iran

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Authors: Sung-Hee Nam, Kwang-Gill Lee, You-Young Jo, HaeYong Kweon

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Entomopathogenic fungi is a Cordyceps species that is isolated from dead silkworm and cicada. Fungi on cicadas were described in old Chinese medicinal books and From ancient times, vegetable wasps and plant worms were widely known to have active substance and have been studied for pharmacological use. Among many fungi belonging to the genus Cordyceps, Cordyceps sinensis have been demonstrated to yield natural products possessing various biological activities and many bioactive components. Generally, It is commonly used to replenish the kidney and soothe the lung, and for the treatment of fatigue. Due to their commercial and economic importance, the demand for Cordyceps has been rapidly increased. However, a supply of Cordyceps specimen could not meet the increasing demand because of their sole dependence on field collection and habitat destruction. Because it is difficult to obtain many insect hosts in nature and the edibility of host insect needs to be verified in a pharmacological aspect. Recently, this setback was overcome that P. tenuipes was able to be cultivated in a large scale using silkworm as host. Pharmacological effects of P. tenuipes cultured on silkworm such as strengthening immune function, anti-fatigue, anti-tumor activity and controlling liver etc have been proved. They are widely commercialized. In this study, we attempted to establish a method for stable growth inhibition of P. tenuipes on silkworm hosts and an optimal condition for synnemata formation. To determine optimum culturing conditions, temperature and light conditions were varied. The length and number of synnemata was highest at 25℃ temperature and 100~300 lux illumination. On an average, the synnemata of wild P. tenuipes measures 70 ㎜ in length and 20 in number; those of the cultured strain were relatively shorter and more in number. The number of synnemata may have increased as a result of inoculating the host with highly concentrated conidia, while the length may have decreased due to limited nutrition per individual. It is not able that changes in light illumination cause morphological variations in the synnemata. However, regulation of only light and temperature could not produce stromata like perithecia, asci, and ascospores. Yamanaka reported that although a complete fruiting body can be produced under optimal culture conditions, it should be regarded as synnemata because it does not develop into an ascoma bearing ascospores.

Keywords: paecilomyces tenuipes, entomopathogenic fungi, silkworm larva, bombyx mori

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Authors: Seongjun Kim, Jinwooung Kim, Sung-Ah Kim

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The development of information and communication technology also affects human cognition and thinking, especially in the field of design, new techniques are being tried. In architecture, new design methodologies such as machine learning or data-driven design are being applied. In particular, these methodologies are used in analyzing the factors related to the value of real estate or analyzing the feasibility in the early planning stage of the apartment housing. However, since the value of apartment buildings is often determined by external factors such as location and traffic conditions, rather than the interior elements of buildings, data is rarely used in the design process. Therefore, although the technical conditions are provided, the internal elements of the apartment are difficult to apply the data-driven design in the design process of the apartment. As a result, the designers of apartment housing were forced to rely on designer experience or modular design alternatives rather than data-driven design at the design stage, resulting in a uniform arrangement of space in the apartment house. The purpose of this study is to propose a methodology to support the designers to design the apartment unit plan with high consumer preference by deriving the correlation and importance of the floor plan elements of the apartment preferred by the consumers through the machine learning and reflecting this information from the early design process. The data on the pre-sale competition rate and the elements of the floor plan are collected as data, and the correlation between pre-sale competition rate and independent variables is analyzed through machine learning. This analytical model can be used to review the apartment unit plan produced by the designer and to assist the designer. Therefore, it is possible to make a floor plan of apartment housing with high preference because it is possible to feedback apartment unit plan by using trained model when it is used in floor plan design of apartment housing.

Keywords: apartment unit plan, data-driven design, design methodology, machine learning

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Authors: D. J. Kalita

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Cancer is one of the prime causes of early death worldwide. Mutation of the gene involve in DNA repair and damage, like BRCA2 (Breast cancer gene two) genes, can be detected efficiently by PCR-RFLP to early breast cancer diagnosis and adopt the suitable method of treatment. Host Defense Peptide can be used as blueprint for the design and synthesis of novel anticancer drugs to avoid the side effect of conventional chemotherapy and chemo resistance. The change at nucleotide position 392 of a -› c in the cancer sample of dog mammary tumour at BRCA2 (exon 7) gene lead the creation of a new restriction site for SsiI restriction enzyme. This SNP may be a marker for detection of canine mammary tumour. Support vector machine (SVM) algorithm was used to design and predict the anticancer peptide from the mature functional peptide. MTT assay of MCF-7 cell line after 48 hours of post treatment showed an increase in the number of rounded cells when compared with untreated control cells. The ability of the synthesized peptide to induce apoptosis in MCF-7 cells was further investigated by staining the cells with the fluorescent dye Hoechst stain solution, which allows the evaluation of the nuclear morphology. Numerous cells with dense, pyknotic nuclei (the brighter fluorescence) were observed in treated but not in control MCF-7 cells when viewed using an inverted phase-contrast microscope. Thus, PCR-RFLP is one of the attractive approach for early diagnosis, and synthetic cationic peptide can be used for the treatment of canine mammary tumour.

Keywords: cancer, cationic peptide, host defense peptides, Breast cancer genes

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

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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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Authors: Ashima Sharma

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Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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Authors: Sajjad Akbar, Rabia Bashir

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With the growth of the number of users, the Internet usage has evolved. Due to its key design principle, there is an incredible expansion in its size. This tremendous growth of the Internet has brought new applications (mobile video and cloud computing) as well as new user’s requirements i.e. content distribution environment, mobility, ubiquity, security and trust etc. The users are more interested in contents rather than their communicating peer nodes. The current Internet architecture is a host-centric networking approach, which is not suitable for the specific type of applications. With the growing use of multiple interactive applications, the host centric approach is considered to be less efficient as it depends on the physical location, for this, Information Centric Networking (ICN) is considered as the potential future Internet architecture. It is an approach that introduces uniquely named data as a core Internet principle. It uses the receiver oriented approach rather than sender oriented. It introduces the naming base information system at the network layer. Although ICN is considered as future Internet architecture but there are lot of criticism on it which mainly concerns that how ICN will manage the most relevant content. For this Web Content Mining(WCM) approaches can help in appropriate data management of ICN. To address this issue, this paper contributes by (i) discussing multiple ICN approaches (ii) analyzing different Web Content Mining approaches (iii) creating a new Internet architecture by merging ICN and WCM to solve the data management issues of ICN. From ICN, Content-Centric Networking (CCN) is selected for the new architecture, whereas, Agent-based approach from Web Content Mining is selected to find most appropriate data.

Keywords: agent based web content mining, content centric networking, information centric networking

Procedia PDF Downloads 473

Authors: Widari

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This study aims to determine the relationship of fast food consumption preferences with macro and micro nutrient adequacy students of SMP Negeri 5 Padang. This study used a cross sectional study conducted on 100 students of SMP Negeri 5 Padang. The variables studied were fast food preferences, nutrition adequacy macronutrients (carbohydrate, protein, fat, fiber) and micro nutrients (sodium, calcium, iron). Confounding factor in this study was the physical activity level because it was considered quite affecting food consumption of students. Data collected by using a questionnaire food recall as many as 2 x 24 hours to see the history of the respondents eat at school day and on holidays. Then, data processed using software Nutrisurvey and Microsoft Excel 2010. The analysis was performed on samples that have low and medium category on physical activity. The physical activity was not analyzed with another variable to see the strength of the relationship between independent and dependent variables. So that, do restrictions on physical activity variables in an attempt to get rid of confounding in design. Univariate and bivariate analyzes performed using SPSS 16.0 for Windows with Kolmogrov-Smirnov statistical tests, confidence level = 95% (α = 0,05). Results of univariate analysis showed that more than 70% of respondents liked fast food. On average, respondents were malnourished macro; malnourished fiber (100%), carbohydrates (72%), and protein (56%), whereas for fat, excess intake of the respondents (41%). Furthermor, many respondents who have micronutrient deficiencies; 98% for sodium, 96% for iron, and 91% for calcium. The results of the bivariate analysis showed no significant association between fast food consumption preferences with macro and micro nutrient adequacy (p > 0,05). This happens because in the fact not all students who have a preference for fast food actually eat them. To study better in the future, it is expected sampling really like and eat fast food in order to obtain better analysis results.

Keywords: fast food, nutritional adequacy, preferences, students

Procedia PDF Downloads 371

Authors: Mohammed Hussein Eltoum Salih

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This study was conducted in four villages, namely: Jadeed, Alandraba, Um Gaar, and EL Shetabe in the White Nile State Sudan, to determine the ecological factors; water vegetations, physical and chemical properties of the water in Snails habitat. Bulinus truncatus, which act as an intermediate host for S. haematobium, were collected from water bodies adjacent to study villages where the residents were suspected to swim, and humans get in contact with water for various purposes. Water samples from the stretches were collected and then measured for parameters that are indicative of the quality of water and sustaining the survival of snails and would confirm even further if the contact between humans and water had taken place. The parameters measured included water conductivity, pH, dissolved oxygen, calcium, and magnesium content. Also, a single water sample from each contact site was collected for microbiological tests. The result revealed that the B. truncatus showed that these animals were fewer and free of infection and their sites of the collection were dense with different plant species making them suitable to harbor snails. Moreover, the results of microbial tests showed that there was higher bacterial contamination. Also, physical and chemical analysis of water sample of contact sites revealed that there were significant differences (p < 0.05) in water pH, calcium, and magnesium content between sites of study villages, and these were discussed in relation to factors suitable for the intermediate hosts and thus for the transmission of the S. haematobium disease.

Keywords: health, parasitology, Schistosoma, snails

Procedia PDF Downloads 145

Authors: Raviporn Madarasmi, Anjalee Vacharaksa, Pravej Serichetaphongse

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The host response in peri-implant tissue may differ from that in periodontal tissue in a healthy individual. The purpose of this study is to investigate the expression of inflammatory cytokines in peri-implant crevicular fluid (PICF) from single implant with different abutment types in comparison to healthy periodontal tissue. 19 participants with healthy implants and teeth were recruited according to inclusion and exclusion criteria. PICF and gingival crevicular fluid (GCF) was collected using sterile paper points. The expression level of inflammatory cytokines including IL-1α, IL-1β, TNF-α, IFN-γ, IL-6, and IL-8 was assessed using enzyme-linked immunosorbent assay (ELISA). Paired t test was used to compare the expression levels of inflammatory cytokines around natural teeth and peri-implant in PICF and GCF of the same individual. The Independent t-test was used to compare the expression levels of inflammatory cytokines in PICF from titanium and UCLA abutment. Expression of IL-6, TNF-α, and IFN-γ in PICF was not statistically different from GCF among titanium and UCLA abutment group. However, the level of IL-1α in the PICF from the implants with UCLA abutment was significantly higher than GCF (P=0.030). In addition, the level of IL-1β in PICF from the implants with titanium abutment was significantly higher than GCF (P=0.032). When different abutment types was compared, IL-8 expression in PICF from implants with UCLA abutment was significantly higher than titanium abutment (P=0.003).

Keywords: abutment, dental implant, gingival crevicular fluid and peri-implant crevicular fluid

Procedia PDF Downloads 183

Authors: S. O. Ayansina, O. A. Adekunle

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There is an increasing demand for a functional extension delivery services in Nigeria with a view to meet up with the food and fiber needs of the ever growing population of human and animal respectively. The study was therefore designed to examine the farmers’ preference for public and private extension services in Southwestern Nigeria, specifically to determine the farmers’ level of participation in the two types of organizations involved and also to evaluate the Performance level of personnel in the two organizations in order to ascertain the beneficiaries’ satisfaction. A multi-stage random sampling technique was used to samples 30 respondents from each of the three selected organizations in each of the three states sampled in Southwestern Nigeria. Hence, 270 respondents were sampled for the study. Data collected were analyzed with Kruskal Wallis one-way Analysis of variance to test the difference between the participation of beneficiaries in the public and private extension services and the level of benefit accrued from the two organizations involved in the study. Results generally revealed that private organizations were performing better and were more preferred by the beneficiaries. Results of the tested hypotheses as shown by Kruskal Wallis test of difference (x2=0.709) indicates no significant difference between farmers’ participation in the extension services of public and private organizations but however shows significant difference (X2=12.074) in the benefits achieved by respondents in the two organizations, such benefits include: increased quantity of Crop produced, farm income, skill acquisition, and improved Education in private extension organizations. Based on this result, it could be inferred that beneficiaries generally preferred private extension organizations because of their effectiveness and vibrancy in programme administration. Public extension is therefore recommended for general overhauling and possibly “merging” of public and private sectors in order to cater for teeming population of farmers demanding for efficient and functional extension services to better their lots both in production and processing.

Keywords: public and private involvement, extension services, farmers’ preferences, Kruskal Wallis Test

Procedia PDF Downloads 331

Authors: Clotilde Guyon, Nada Jmari, Yen-Chin Li, Jean Denoyel, Noriyuki Fujikado, Christophe Blanchet, David Root, Matthieu Giraud

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Aire induces ectopic expression of a large repertoire of tissue-specific antigen (TSA) genes in thymic medullary epithelial cells (MECs), driving immunological self-tolerance in maturing T cells. Although important mechanisms of Aire-induced transcription have recently been disclosed through the identification and the study of Aire’s partners, the fine transcriptional functions underlied by a number of them and conferred to Aire are still unknown. Alternative cleavage and polyadenylation (APA) is an essential mRNA processing step regulated by the termination complex consisting of 85 proteins, 10 of them have been related to Aire. We evaluated APA in MECs in vivo by microarray analysis with mRNA-spanning probes and RNA deep sequencing. We uncovered the preference of Aire-dependent transcripts for short-3’UTR isoforms and for proximal poly(A) site selection marked by the increased binding of the cleavage factor Cstf-64. RNA interference of the 10 Aire-related proteins revealed that Clp1, a member of the core termination complex, exerts a profound effect on short 3’UTR isoform preference. Clp1 is also significantly upregulated in the MECs compared to 25 mouse tissues in which we found that TSA expression is associated with longer 3’UTR isoforms. Aire-dependent transcripts escape a global 3’UTR lengthening associated with MEC differentiation, thereby potentiating the repressive effect of microRNAs that are globally upregulated in mature MECs. Consistent with these findings, RNA deep sequencing of actinomycinD-treated MECs revealed the increased stability of short 3’UTR Aire-induced transcripts, resulting in TSA transcripts accumulation and contributing for their enrichment in the MECs.

Keywords: Aire, central tolerance, miRNAs, transcription termination

Procedia PDF Downloads 381

Authors: Rosa Suryantini, Reine S. Wulandari, Slamet Rifanjani

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Acacia (Acacia mangium) is a forest plant species which is produced to pulp and paper. The high demand for pulp and paper increase the acacia plantation forest area. However, the outbreak of Ganoderma (root rot pathogen) infection becomes obstacles for the development of acacia plantations. This is due to the extent of host range and species of Ganoderma. Ganoderma has also the ability to survive the long-term without hosts. The diversity of the host and Ganoderma species affects its virulence. Therefore, this study aimed to determine the virulence of Ganoderma from different hosts (acacia, palm oil (Elaeis guineensis) and rubber (Hevea brasiliensis)). The methods were isolation and morphology identification of Ganoderma, and inoculation of Ganoderma isolates on acacia seedlings. The results showed that the three isolates of Ganoderma from different hosts had a morphological similarity with G. Lucidum (according to Ganoderma isolated from acacia or G1), G. boninense (according to Ganoderma isolated from palm oil or G2) and G. applanatum (according to Ganoderma isolated from rubber or G3). Symptoms of infection in acacia were seen at 3 months of age. The symptoms were begun with chlorosis, necrosis and death of seedlings (such as burning). Necrosis was started from the tip of the leaf. Based on this visible symptoms, G1 was moderate virulence isolate and G2 was low virulence isolate while G3 was avirulen isolate. The symptoms were still growing in accordance with the development of plant so it affected the value of diseases severity index. Ganoderma infection decreased the dry weight of seedlings, ie. 3.82 g (seedlings that were inoculated by G1), 4.01 g (seedlings that were inoculated by G2); and 5.02 g (seedlings that were inoculated by G3) when the dry weight of seedlings control was 10,02 g. These results provide information for early control of Ganoderma diseases on acacia especially those planted near rubber and oil palm crops.

Keywords: Acacia, Ganoderma, infection, virulence

Procedia PDF Downloads 190

Authors: Tanja Kreitenweis

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Lifelong learning is increasingly seen as essential for coping with the rapidly changing work environment. To this end, serious games can provide convenient and straightforward access to complex knowledge for all age groups. However, learning achievements depend largely on a learner’s non-cognitive skill disposition (e.g., motivation, self-belief, playfulness, and openness). With the aim of combining the fields of serious games and non-cognitive skills, this research focuses in particular on the use of a business simulation, which conveys change management insights. Business simulations are a subset of serious games and are perceived as a non-traditional learning method. The presented objectives of this work are versatile: (1) developing a scale, which measures learners’ knowledge and skills level before and after a business simulation was played, (2) investigating the influence of non-cognitive skills on learning in this business simulation environment and (3) exploring the moderating role of team preference in this type of learning setting. First, expert interviews have been conducted to develop an appropriate measure for learners’ skills and knowledge assessment. A pretest-posttest experimental design with German management students was implemented to approach the remaining objectives. By using the newly developed, reliable measure, it was found that students’ skills and knowledge state were higher after the simulation had been played, compared to before. A hierarchical regression analysis revealed two positive predictors for this outcome: motivation and self-esteem. Unexpectedly, playfulness had a negative impact. Team preference strengthened the link between grit and playfulness, respectively, and learners’ skills and knowledge state after completing the business simulation. Overall, the data underlined the potential of business simulations to improve learners’ skills and knowledge state. In addition, motivational factors were found as predictors for benefitting most from the applied business simulation. Recommendations are provided for how pedagogues can use these findings.

Keywords: business simulations, change management, (experiential) learning, non-cognitive skills, serious games

Procedia PDF Downloads 104

Authors: Sara Dalma Galgoczi, Judith Gabriella Kengyel

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Sex scandals are quite an engaging topic to work with, especially with their judgment in society. Most people are interested in other people's lives, specifically in public figures' such as celebrities or politicians, because ordinary people feel like they have the right to know more things about the famous and notorious ones than they would probably willing to share. Intimacy and sexual acts aren't exceptions; moreover, sexuality is one of the central interests of humans ever since. Besides, knowing and having an opinion about any kind of scandal can change even whole social groups or classes estimation of anyone. This study aims to research the social judgment of some Hungarian politicians' sex scandals and asks important questions like diverse public opinions in the light of gender or delegates’ abuse of power. Considering that this study is about collecting and evaluating opinions from the public, and no one before researched and published this exact topic and cases, an online survey was created. In the survey were different sections. We collected data about party-preference, conservativism-liberalism scale; then we used the following questionnaires: from Zero-sum perspective with regard to gender equality (Ruthig, Kehn, Gamblin, Vanderzanden & Jones, 2017), Ambivalent Sexism Inventory (ASI; Glick & Fiske, 1996), Ambivalence Toward Men Inventory (AMI; Glick & Fiske, 1999). Finally, 5 short summaries were presented about five Hungarian politicians' sex scandal cases (3 males, 2 females) from the recent past. These stories were followed by questions about their opinion of the party and attitudes towards the parties' reactions to the cases. We came to the conclusion that people are more permissive with the scandals of men, and benevolent sexism and ambivalence towards men mediate this relation. Men tend to see these cases as part of politicians' private lives more than women. Party preference had a significant effect - people tend to pass a sentence the delegates of the opposing parties, and they rather release the delegates of their preferred party.

Keywords: sex scandal, sexism, social judgement, politician

Procedia PDF Downloads 121

Authors: D. Zahran, M. AlKhazindar, M. Khalil, E. T. A. Sayed

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Streptomyces scabies is a pathogenic actinomycete that infects potato crop causing severe production losses. Actinophages affect the composition and diversity of the bacterial population, thereby, can be used as a biological control. Samples of actinomycetes and phages were collected from different cultivated soils including farms of Faculty of Science, Faculty of Agriculture and different locations in Giza, Egypt. Actinomycetes were identified by using biochemical, morphological tests and molecular studies using 16S rRNA sequencing. Two specific phages (E1 and E2) against Streptomyces scabies and other hosts were isolated. Phages were identified using dilution end point (DEP), longevity in vitro (LIV), thermal inactivation point (TIP), host range and electron microscopy. PhageE1 was characterized by 10-8 (DEP),180 days(LIV), 95°C(TIP), narrow host range and electron microscopy showed ahead (59.9 nm) and neck (10.4nm). On the other hand phageE2 had 10-20 (DEP),180 days(LIV), 90°C(TIP), and the size of head was (67.2 nm) and tail (114nm). Antiviral activity was also studied using different chemicals (NaCL, KCL, CaCL2, BaCL2, CoCL2, AgNO3, ALCL3and HgCL2) with different concentrations and different plant extracts with different concentrations (star anise, tea, tillia, peppermint, ginger, cumin, chamomile, turmeric cinnamon, marjoram and black cumin). Both Phage E1and phage E2 were vulnerable to (cumin, ginger, chamomile, guavas leaves and star anise) but resistant to (Tillie, marjoram, fennelflower seeds, peppermint, and cinnamon).

Keywords: potato scab, actinophages, biological control, electron microscopy, TIP, DEP, LIV, antiviral activity

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Authors: Vrushali Guhe, Shailza Singh

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Cutaneous leishmaniasis is neglected tropical disease present worldwide caused by the protozoan parasite Leishmania major, the therapeutic armamentarium for leishmaniasis are showing several limitations as drugs are showing toxic effects with increasing resistance by a parasite. Thus identification of novel therapeutic targets is of paramount importance. Previous studies have shown that autophagy, a cellular process, can either facilitate infection or aid in the elimination of the parasite, depending on the specific parasite species and host background in leishmaniasis. In the present study, our objective was to target the essential autophagy protein ATG8, which plays a crucial role in the survival, infection dynamics, and differentiation of the Leishmania parasite. ATG8 in Leishmania major and its homologue, LC3, in Homo sapiens, act as autophagic markers. Present study manifested the crucial role of ATG8 protein as a potential target for combating Leishmania major infection. Through bioinformatics analysis, we identified non-conserved motifs within the ATG8 protein of Leishmania major, which are not present in LC3 of Homo sapiens. Against these two non-conserved motifs, we generated a peptide library of 60 peptides on the basis of physicochemical properties. These peptides underwent a filtering process based on various parameters, including feasibility of synthesis and purification, compatibility with Selective Reaction Monitoring (SRM)/Multiple reaction monitoring (MRM), hydrophobicity, hydropathy index, average molecular weight (Mw average), monoisotopic molecular weight (Mw monoisotopic), theoretical isoelectric point (pI), and half-life. Further filtering criterion shortlisted three peptides by using molecular docking and molecular dynamics simulations. The direct interaction between ATG8 and the shortlisted peptides was confirmed through Surface Plasmon Resonance (SPR) experiments. Notably, these peptides exhibited the remarkable ability to penetrate the parasite membrane and exert profound effects on Leishmania major. The treatment with these peptides significantly impacted parasite survival, leading to alterations in the cell cycle and morphology. Furthermore, the peptides were found to modulate autophagosome formation, particularly under starved conditions, suggesting their involvement in disrupting the regulation of autophagy within Leishmania major. In vitro, studies demonstrated that the selected peptides effectively reduced the parasite load within infected host cells. Encouragingly, these findings were corroborated by in vivo experiments, which showed a reduction in parasite burden upon peptide administration. Additionally, the peptides were observed to affect the levels of LC3II within host cells. In conclusion, our findings highlight the efficacy of these novel peptides in targeting Leishmania major’s ATG8 and disrupting parasite survival. These results provide valuable insights into the development of innovative therapeutic strategies against leishmaniasis via targeting autophagy protein ATG8 of Leishmania major.

Keywords: ATG8, leishmaniasis, surface plasmon resonance, MD simulation, molecular docking, peptide designing, therapeutics

Procedia PDF Downloads 78

Authors: Dagmara A. Niedziela, Mark P. Murphy, Orla M. Keane, Finola C. Leonard

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Staphylococcus aureus is the most frequent cause of clinical and subclinical bovine mastitis in Ireland. Mastitis caused by S. aureus is often chronic and tends to recur after antibiotic treatment. This may be due to several virulence factors, including attributes that enable the bacterium to internalize into bovine mammary epithelial cells, where it may evade antibiotic treatment, or evade the host immune response. Four bovine-adapted lineages (CC71, CC97, CC151 and ST136) were identified among a collection of Irish S. aureus mastitis isolates. Genotypic variation of mastitis-causing strains may contribute to different presentations of the disease, including differences in milk somatic cell count (SCC), the main method of mastitis detection. The objective of this study was to investigate the influence of bacterial strain and lineage on host immune response, by employing cell culture methods in vitro as well as an in vivo infection model. Twelve bovine adapted S. aureus strains were examined for internalization into bovine mammary epithelial cells (bMEC) and their ability to induce an immune response from bMEC (using qPCR and ELISA). In vitro studies found differences in a variety of virulence traits between the lineages. Strains from lineages CC97 and CC71 internalized more efficiently into bovine mammary epithelial cells (bMEC) than CC151 and ST136. CC97 strains also induced immune genes in bMEC more strongly than strains from the other 3 lineages. One strain each of CC151 and CC97 that differed in their ability to cause an immune response in bMEC were selected on the basis of the above in vitro experiments. Fourteen first-lactation Holstein-Friesian cows were purchased from 2 farms on the basis of low SCC (less than 50 000 cells/ml) and infection free status. Seven cows were infected with 1.73 x 102 c.f.u. of the CC97 strain (Group 1) and another seven with 5.83 x 102 c.f.u. of the CC151 strain (Group 2). The contralateral quarter of each cow was inoculated with PBS (vehicle). Clinical signs of infection (temperature, milk and udder appearance, milk yield) were monitored for 30 days. Blood and milk samples were taken to determine bacterial counts in milk, SCC, white blood cell populations and cytokines. Differences in disease presentation in vivo between groups were observed, with two animals from Group 2 developing clinical mastitis and requiring antibiotic treatment, while one animal from Group 1 did not develop an infection for the duration of the study. Fever (temperature > 39.5⁰C) was observed in 3 animals from Group 2 and in none from Group 1. Significant differences in SCC and bacterial load between groups were observed in the initial stages of infection (week 1). Data is also being collected on cytokines and chemokines secreted during the course of infection. The results of this study suggest that a strain from lineage CC151 may cause more severe clinical mastitis, while a strain from lineage CC97 may cause mild, subclinical mastitis. Diversity between strains of S. aureus may therefore influence the clinical presentation of mastitis, which in turn may influence disease detection and treatment needs.

Keywords: Bovine mastitis, host immune response, host-pathogen interactions, Staphylococcus aureus

Procedia PDF Downloads 156

Authors: Santi Maneewatchararangsri, Wanpen Chaicumpa, Patcharin Saengjaruk, Urai Chaisri

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Leptospirosis is a globally neglected disease that continues to be a significant public health and veterinary burden, with millions of cases reported each year. Early and accurate differential diagnosis of leptospirosis from other febrile illnesses and the development of a broad spectrum of leptospirosis vaccines are needed. The LipL32 outer membrane lipoprotein is a member of Leptospira adhesive matrices and has been found to exert hemolytic activity to erythrocytes in vitro. Therefore, LipL32 is regarded as a potential target for diagnosis, broad-spectrum leptospirosis vaccines, and for passive immunotherapy. In this study, we established LipL32-specific mouse monoclonal antibodies, mAbLPF1 and mAbLPF2, and their respective mouse- and humanized-engineered single chain variable fragment (ScFv). Their antibodies’ neutralizing activities against Leptospira-mediated hemolysis in vitro, and the therapeutic efficacy of mAbs against heterologous Leptospira infected hamsters were demonstrated. The epitope peptide of mAb LPF1 was mapped to a non-contiguous carboxy-terminal β-turn and amphipathic α-helix of LipL32 structure contributing to phospholipid/host cell adhesion and membrane insertion. We found that the mAbLPF2 epitope was located on the interacting loop of peptide binding groove of the LipL32 molecule responsible for interactions with host constituents. Epitope sequences are highly conserved among Leptospira spp. and are absent from the LipL32 superfamily of other microorganisms. Both epitopes are surface-exposed, readily accessible by mAbs, and immunogenic. However, they are less dominant when revealed by LipL32-specific immunoglobulins from leptospirosis-patient sera and rabbit hyperimmune serum raised by whole Leptospira. Our study also demonstrated an adhesion inhibitory activity of LipL32 protein to host membrane components and cells mediated by mAbs as well as an anti-hemolytic activity of the respective antibodies. The therapeutic antibodies, particularly the humanized-ScFv, have a potential for further development as non-drug therapeutic agent for human leptospirosis, especially in subjects allergic to antibiotics. The epitope peptides recognized by two therapeutic mAbs have potential use as tools for structure-function studies. Finally, protective peptides may be used as a target for epitope-based vaccines for control of leptospirosis.

Keywords: leptospira lipl32-specific antibodies, therapeutic epitopes, epitopes characterization, immunotherapy

Procedia PDF Downloads 297