Search results for: snail host
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 925

Search results for: snail host

745 Exploring Labor Market Participation of Highly Skilled Immigrant Women in the United States: Barriers and Strategies

Authors: Yurdum Cokadar

Abstract:

The United States is the country where the majority of highly skilled immigrants are hosted. Two-thirds of foreign-born migrants from Turkey - an underrepresented and understudied immigrant group in the United States - are highly skilled. Generated by the aim of filling this gap in the literature, the motivation of this research is to understand highly skilled Turkish immigrant women’s integration into the U.S. labor market, including barriers that they face and strategies they develop to rebuild their career after relocation. The in-depth interviews of 20 highly skilled Turkish women residing in the U.S. revealed that the majority of women participants are either not integrated into the labor market, occupy positions below their skill, or cannot reach the same upper segments of the labor market in the host country, arising from a range of structural and personal barriers interplaying in their career trajectories. Furthermore, many of them cannot transfer their social and cultural capital gained in their home country into the United States. The labor market participation process of these women is analyzed in the light of Bourdieu’s theory of capital and the intersectional approach of gender, class and ethnicity in order to understand the positions of highly skilled immigrant women in the host country labor market.

Keywords: deskilling, gender, class and ethnicity, highly skilled women immigrants, integration into the U.S. the labor market, labor market participation, skilled migration, theory of capital

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744 Pyramidal Lucas-Kanade Optical Flow Based Moving Object Detection in Dynamic Scenes

Authors: Hyojin Lim, Cuong Nguyen Khac, Yeongyu Choi, Ho-Youl Jung

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In this paper, we propose a simple moving object detection, which is based on motion vectors obtained from pyramidal Lucas-Kanade optical flow. The proposed method detects moving objects such as pedestrians, the other vehicles and some obstacles at the front-side of the host vehicle, and it can provide the warning to the driver. Motion vectors are obtained by using pyramidal Lucas-Kanade optical flow, and some outliers are eliminated by comparing the amplitude of each vector with the pre-defined threshold value. The background model is obtained by calculating the mean and the variance of the amplitude of recent motion vectors in the rectangular shaped local region called the cell. The model is applied as the reference to classify motion vectors of moving objects and those of background. Motion vectors are clustered to rectangular regions by using the unsupervised clustering K-means algorithm. Labeling method is applied to label groups which is close to each other, using by distance between each center points of rectangular. Through the simulations tested on four kinds of scenarios such as approaching motorbike, vehicle, and pedestrians to host vehicle, we prove that the proposed is simple but efficient for moving object detection in parking lots.

Keywords: moving object detection, dynamic scene, optical flow, pyramidal optical flow

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743 Production of Novel Antibiotics of Tylosin by Importing eryK and eryG Genes in Streptomyces fradiae

Authors: Neda Gegar Goshe, M. Moradi, Hossein Rassi

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The antibacterial properties of macrolide antibiotics (such as erythromycin and tylosin) depend ultimately on the glycosylation of otherwise inactive polyketide lactones. Among the sugars commonly found in such macrolides are various 6-deoxyhexoses including the 3-dimethylamino sugars mycaminose and desosamine (4-deoxymycaminose). Some macrolides (such as tylosin) possess multiple sugar moieties, whereas others (such as erythromycin) have two sugar substituents. Streptomyces fradiae is an ideal host for development of generic polyketide-overproducing strains because it contains three of the most common precursors-malonyl-CoA, methylmalonyl-CoA and ethylmalonyl-CoA-used by modular PKS, and is a host that is amenable to genetic manipulation. As patterns of glycosylation markedly influence a macrolide's drug activity, there is considerable interest in the possibility of using combinatorial biosynthesis to generate new pairings of polyketide lactones with sugars, especially 6-deoxyhexoses. Here, we report a successful attempt to alter the aminodeoxyhexose-biosynthetic capacity of Streptomyces fradiae (a producer of tylosin) by importing genes from the erythromycin producer Saccharopolyspora erythraea. The bio transformation of erythromycin-D into the desired major component erythromycin-A involves two final enzymatic reactions, EryK-catalyzed hydroxylation at the C-12 position of the aglycone and EryG-catalyzed O methylation at the C-3 position of macrose. This engineered S. fradiae produced substantial amounts of two potentially useful macrolides that had not previously been obtained by fermentation.

Keywords: tylosin, eryK and eryG genes, streptomyces fradiae

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742 An in Situ Dna Content Detection Enabled by Organic Long-persistent Luminescence Materials with Tunable Afterglow-time in Water and Air

Authors: Desissa Yadeta Muleta

Abstract:

Purely organic long-persistent luminescence materials (OLPLMs) have been developed as emerging organic materials due to their simple production process, low preparation cost and better biocompatibilities. Notably, OLPLMs with afterglow-time-tunable long-persistent luminescence (LPL) characteristics enable higher-level protection applications and have great prospects in biological applications. The realization of these advanced performances depends on our ability to gradually tune LPL duration under ambient conditions, however, the strategies to achieve this are few due to the lack of unambiguous mechanisms. Here, we propose a two-step strategy to gradually tune LPL duration of OLPLMs over a wide range of seconds in water and air, by using derivatives as the guest and introducing a third-party material into the host-immobilized host–guest doping system. Based on this strategy, we develop an analysis method for deoxyribonucleic acid (DNA) content detection without DNA separation in aqueous samples, which circumvents the influence of the chromophore, fluorophore and other interferents in vivo, enabling a certain degree of in situ detection that is difficult to achieve using today’s methods. This work will expedite the development of afterglow-time-tunable OLPLMs and expand new horizons for their applications in data protection, bio-detection, and bio-sensing

Keywords: deoxyribonucliec acid, long persistent luminescent materials, water, air

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741 Potential Role of Arbuscular Mycorrhizal (AM) Fungi in CO₂-Sequestration During Bipartite Interaction with Host Plant Oryza Sativa

Authors: Sadhana Shukla, Pushplata Singh, Nidhi Didwania

Abstract:

Arbuscular mycorrhizal (AM) fungi are a highly advantageous and versatile group of fungi that significantly contribute to the formation of soil organic matter by creating a demand for plant carbon (C) and distributing it through below-ground hyphal biomass, regardless of their substantial contribution in enhancing net primary productivity and accumulating additional photosynthetic fixed C in the soil. The genetic role of AM fungi in carbon cycling is largely unexplored. In our study, we propose that AM fungi significantly interact with the soil, particularly: the provision of photosynthates by plants. We have studied the expression of AM fungi genes involved in CO₂ sequestration during host-plant interaction was investigated by qPCR studies. We selected Rhizophagus proliferus (AM fungi) and Oryza sativa (Rice) (inoculated with or without 200ppg AMF inoculums per plant) and investigated the effect of AM fungi on soil organic carbon (SOC) and rice growth under field conditions. Results thus provided faster SOC turnover, 35% increased nutrient uptake in plants and pronounced hyphal biomass of AM fungi which enhanced soil carbon storage by 15% in comparison to uninoculated plants. This study will offer a foundation for delving into various carbon-soil studies while also advancing our comprehension of the relationship between AM fungi and the sustainability of agricultural ecosystems.

Keywords: arbuscular mycorrhizal (AM) fungi, carbon sequestration, gene expression, soil health, plant development.

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740 Influence of Season, Temperature, and Photoperiod on Growth of the Land Snail Helix aperta

Authors: S. Benbellil-Tafoughalt, J. M. Koene

Abstract:

Growth strategies are often plastic and influenced by environmental conditions. Terrestrial gastropods are particularly affected by seasonal and climatic variables, and growth rate and size at maturity are key traits in their life history. Therefore, we investigated juvenile growth of Helix aperta snails under four combinations of temperature and photoperiod using two sets of young snails, born in the laboratory from adults collected in either the autumn (aestivating snails) or spring (active snails). Parental snails were collected from Bakaro (Northeastern Algeria). Higher temperature increased adult size and reduced time to reproduction. Long day photoperiod also increased the final body weight, but had no effect on the length of the growth period. The season of birth had significant effects on length of the growth period and weight of hatchings, whereas this weight difference disappeared by adulthood. The spring snails took less time to develop and reached similar adult body weight as the autumn snails. These differences may be due to differences in egg size or quality between the snails from different seasons. More rapid growth in spring snails results in larger snails entering aestivation, a period with size-related mortality in this species.

Keywords: growth, Hélix aperta, photoperiod, temperature

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739 Proximate Analysis of Muscle of Helix aspersa Living in Konya, Turkey

Authors: Ozcan Baris Citil

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The aim of the present study is the determination of the effects of variations in the proximate analysis, cholesterol content and fatty acid compositions of Helix aspersa. Garden snails (Helix aspersa) were picked up by hand from the Central Anatolia Region of Turkey, in autumn (November) in 2015. Fatty acid methyl esters (FAMEs) and cholesterol analysis were analyzed by gas chromatography (GC). The protein contents of snail muscle were determined with Kjeldahl distillation units. Statistical comparisons were made by using SPSS Software (version 16.0). Thirty different fatty acids of different saturation levels were detected. As the predominant fatty acids, stearic acid (C18:0), oleic acid (C18:1ω9), linoleic acid (C18:2ω6), palmitic acid (C16:0), arachidonic acid (C20:4ω6), eicosadienoic acid (C20:2) and linolenic acid (C18:3ω3) were found in Helix aspersa. Palmitic acid (C16:0) was identified as the major SFA in autumn. Linoleic acid (C18:2ω6), eicosadienoic acid (C20:2) and arachidonic acid (C20:4ω6) have the highest levels among the PUFAs. In the present study, ω3 were found 5.48% in autumn. Linolenic acid and omega-3 fatty acid amounts in the autumn decreased significantly but cholesterol content was not affected in Helix aspersa in autumn (November) in 2015.

Keywords: Helix aspersa, fatty acid, SFA, PUFA, cholesterol

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738 Technology Transfer and FDI: Some Lessons for Tunisia

Authors: Assaad Ghazouani, Hedia Teraoui

Abstract:

The purpose of this article is to try to see if the FDI actually contributes to technology transfer in Tunisia or are there other sources that can guarantee this transfer? The answer to this problem was gradual as we followed an approach using economic theory, the reality of Tunisia and econometric and statistical tools. We examined the relationship between technology transfer and FDI in Tunisia over a period of 40 years from 1970 to 2010. We estimated in two stages: first, a growth equation, then we have learned from this regression residue (proxy technology), secondly, we regressed on European FDI, exports of manufactures, imports of goods from the European Union in addition to other variables to test the robustness of the results and describing the level of infrastructure in the country. It follows from our study that technology transfer does not originate primarily and exclusively in the FDI and the latter is econometrically weakly with technology transfer and spill over effect of FDI does not seem to occur according to our results. However, the relationship between technology transfer and imports is negative and significant. Although this result is cons-intuitive, is recurrent in the literature of panel data. It has also given rise to intense debate on the microeconomic modelling as well as on the empirical applications. Technology transfer through trade or foreign investment has become a catalyst for growth recognized by numerous empirical studies in particular. However, the relationship technology transfer FDI is more complex than it appears. This complexity is due, primarily, but not exclusively to the close link between FDI and the characteristics of the host country. This is essentially the host's responsibility to establish general conditions, transparent and conducive to investment, and to strengthen human and institutional capacity necessary for foreign capital flows that can have real effects on growth.

Keywords: technology transfer, foreign direct investment, economics, finance

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737 Sorption of Crystal Violet from Aqueous Solution Using Chitosan−Charcoal Composite

Authors: Kingsley Izuagbe Ikeke, Abayomi O. Adetuyi

Abstract:

The study investigated the removal efficiency of crystal violet from aqueous solution using chitosan-charcoal composite as adsorbent. Deproteination was carried out by placing 200g of powdered snail shell in 4% w/v NaOH for 2hours. The sample was then placed in 1% HCl for 24 hours to remove CaCO3. Deacetylation was done by boiling in 50% NaOH for 2hours. 10% Oxalic acid was used to dissolve the chitosan before mixing with charcoal at 55°C to form the composite. The composite was characterized by Fourier Transform Infra-Red and Scanning Electron Microscopy measurements. The efficiency of adsorption was evaluated by varying pH of the solution, contact time, initial concentration and adsorbent dose. Maximum removal of crystal violet by composite and activated charcoal was attained at pH10 while maximum removal of crystal violet by chitosan was achieved at pH 8. The results showed that adsorption of both dyes followed the pseudo-second-order rate equation and fit the Langmuir and Freundlich isotherms. The data showed that composite was best suited for crystal violet removal and also did relatively well in the removal of alizarin red. Thermodynamic parameters such as enthalpy change (ΔHº), free energy change (ΔGº) and entropy change (ΔSº) indicate that adsorption process of Crystal Violet was endothermic, spontaneous and feasible respectively.

Keywords: crystal violet, chitosan−charcoal composite, extraction process, sorption

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736 Molecular Implication of Interaction of Human Enteric Pathogens with Phylloplane of Tomato

Authors: Shilpi, Indu Gaur, Neha Bhadauria, Susmita Goswami, Prabir K. Paul

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Cultivation and consumption of organically grown fruits and vegetables have increased by several folds. However, the presence of Human Enteric Pathogens on the surface of organically grown vegetables causing Gastro-intestinal diseases, are most likely due to contaminated water and fecal matter of farm animals. Human Enteric Pathogens are adapted to colonize the human gut, and also colonize plant surface. Microbes on plant surface communicate with each other to establish quorum sensing. The cross talk study is important because the enteric pathogens on phylloplane have been reported to mask the beneficial resident bacteria of plant. In the present study, HEPs and bacterial colonizers were identified using 16s rRNA sequencing. Microbial colonization patterns after interaction between Human Enteric Pathogens and natural bacterial residents on tomato phylloplane was studied. Tomato plants raised under aseptic conditions were inoculated with a mixture of Serratia fonticola and Klebsiella pneumoniae. The molecules involved in cross-talk between Human Enteric Pathogens and regular bacterial colonizers were isolated and identified using molecular techniques and HPLC. The colonization pattern was studied by leaf imprint method after 48 hours of incubation. The associated protein-protein interaction in the host cytoplasm was studied by use of crosslinkers. From treated leaves the crosstalk molecules and interaction proteins were separated on 1D SDS-PAGE and analyzed by MALDI-TOF-TOF analysis. The study is critical in understanding the molecular aspects of HEP’s adaption to phylloplane. The study revealed human enteric pathogens aggressively interact among themselves and resident bacteria. HEPs induced establishment of a signaling cascade through protein-protein interaction in the host cytoplasm. The study revealed that the adaptation of Human Enteric Pathogens on phylloplane of Solanum lycopersicum involves the establishment of complex molecular interaction between the microbe and the host including microbe-microbe interaction leading to an establishment of quorum sensing. The outcome will help in minimizing the HEP load on fresh farm produce, thereby curtailing incidences of food-borne diseases.

Keywords: crosslinkers, human enteric pathogens (HEPs), phylloplane, quorum sensing

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735 Role of Vitamin D in Osseointegration of Dental Implant

Authors: Pouya Khaleghi

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Dental implants are a successful treatment modality for restoring both function and aesthetics. Dental implant treatment has predictive results in the replacement of the lost teeth and has a high success rate even in the long term. The most important factor which is responsible for the positive course of implant treatment is the process of osseointegration between the implant structure and the host’s bone tissue. During recent years, many studies have focused on surgical and prosthetic factors, as well as the implant-related factors. However, implant failure still occurs despite the improvements that have led to the increased survival rate of dental implants, which suggests the possible role of some host-related risk factors. Vitamin D is a fat-soluble vitamin regulating calcium and phosphorus metabolism in tissues. The role of vitamin D in bone healing has been under investigation for several years. Vitamin D deficiency has also been associated with impaired and delayed callus formation and fractures healing; however, the role of vitamin D has not been clarified. Therefore, it is extremely important to study the phenomenon of a connection formed between bone tissue and the surface of a titanium implant and find correlations between the 25- hydroxycholecalciferol concentration in blood serum and the course of osseointegration. Because the processes of bone remodeling are very dynamic in the period of actual osseointegration, it is necessary to obtain the correct concentration of vitamin D3 metabolites in blood serum. In conclusion, the correct level of 25-hydroxycholecalciferol on the day of surgery and vitamin D deficiency treatment have a significant influence on the increase in the bone level at the implant site during the process of osseointegration assessed radiologically.

Keywords: implant, osseointegration, vitamin d, dental

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734 The Role of Glutamine-Rich Region of Candida Albicans Tec1p in Mediating Morphological Transition and Invasive Growth

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

Abstract:

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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733 Economic Impact of Ogbomoso Migrant Community in Jos Metropolis, Nigeria, 1940-2000

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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732 Influence of People and Places on the Identity of Ethnic Enclaves: A Visual Analysis of Little India, Penang

Authors: Excellent Hansda

Abstract:

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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731 The Experience of Community-based Tourism in Yunguilla, Ecuador and Its Social-Cultural Impact

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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730 First-Principles Calculations of Hydrogen Adsorbed in Multi-Layer Graphene

Authors: Mohammad Shafiul Alam, Mineo Saito

Abstract:

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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729 Remittances, Unemployement and Demographic Changes between Tunisia and Europe

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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728 Analysis of the Impact of Foreign Direct Investment on the Integration of the Automotive Industry of Iran into Global Production Networks

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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727 Optimization of Culture Conditions of Paecilomyces Tenuipes, Entomopathogenic Fungi Inoculated into the Silkworm Larva, Bombyx Mori

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

Abstract:

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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726 Effect of Silicon on Tritrophic Interaction of Cotton, Whitefly and Chrysoperla carnea

Authors: Asim Abbasi, Muhammad Sufyan

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The present experiment was carried out to examine the effects of silicon dioxide on tritrophic interaction of cotton, whitefly, and the predator Chrysoperla carnea. Population of whitefly was maintained on silicon treated and non-treated cotton for two generations in greenhouse net cages exposed to outside temperature and luminosity. The cotton was treated with silicon dioxide twice after 15 days intervals with 200 ppm concentration. A stock rearing of the natural predator was developed in the laboratory conditions. In the bioassay eggs of the predator all at the same age were individualized in glass petri plates that will be pierced with a pin to allow aeration and maintained in an incubator at 28 ± 2°C, 70 ± 10% relative humidity and 12h photo phase. Population of whitefly stayed on silicon treated, and non-treated cotton were offered to newly hatched chrysopid larvae until the end of the larval stage, assuring a permanent supply. Feeding preference of C. carnea along with longevity, survival of each instar larvae, pupation, adult emergence, and fecundity was checked. The results revealed that there was no significant difference in the feeding preference of C. carnea among both treatments. Durations of 1st and 2nd larval instar were also at par in both treatments. However overall longevity and adult emergence were a bit lower in silicon treated whitefly treatment. This may be due to the fact that silicon reduces the nutritional quality of host because of reduced whitefly feeding on silicon treated cotton. No significant difference in 1st and 2nd larval instars and then increased larval duration in later instars suggested that the effect of silicon treated host should be checked on more than 1 generation of C. carnea to get better findings.

Keywords: Chrysoperla carnea, silicon, tritrophic, whitefly

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725 Early Diagnosis and Treatment of Cancer Using Synthetic Cationic Peptide

Authors: D. J. Kalita

Abstract:

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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724 A 3d Intestine-On-Chip Model Allows Colonization with Commensal Bacteria to Study Host-Microbiota Interaction

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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723 Enhanced Functional Production of a Crucial Biomolecule Human Serum Albumin in Escherichia coli

Authors: Ashima Sharma

Abstract:

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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722 Emergence of Information Centric Networking and Web Content Mining: A Future Efficient Internet Architecture

Authors: Sajjad Akbar, Rabia Bashir

Abstract:

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

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721 Host Responses in Peri-Implant Tissue in Comparison to Periodontal Tissue

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

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720 Ganoderma Infection in Acacia mangium: Difference of Plant Hosts to Virulency of Ganoderma

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

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719 Isolation and Characterization of Actinophages Infecting Streptomyces scabies in Egypt

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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718 Targeting Peptide Based Therapeutics: Integrated Computational and Experimental Studies of Autophagic Regulation in Host-Parasite Interaction

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

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717 Clinical Presentation and Immune Response to Intramammary Infection of Holstein-Friesian Heifers with Isolates from Two Staphylococcus aureus Lineages

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

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716 Leptospira Lipl32-Specific Antibodies: Therapeutic Property, Epitopes Characterization and Molecular Mechanisms of Neutralization

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

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