Search results for: resistant gene expression

Authors: Mohamad Mohsen Homayouni, Niloofar Taghipour, Ahmad Reza Memar, Niloofar Khalaji, Hamed Kiani, Seyyed Javad Seyyed Tabaei

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Background and Objective: Cryptosporidium is a coccidian protozoan parasite; its oocysts in surface water are a global health problem. Due to the low number of parasites in the water resources and the lack of laboratory culture, rapid and sensitive method for detection of the organism in the water resources is necessarily required. We applied modified acid-fast staining and PCR for the detection of the Cryptosporidium spp. and analysed the genotypes in 55 samples collected from surface water. Methods: Over a period of nine months, 55 surface water samples were collected from the five rivers in Tehran, Iran. The samples were filtered by using cellulose acetate membrane filters. By acid fast method, initial identification of Cryptosporidium oocyst were carried out on surface water samples. Then, nested PCR assay was designed for the specific amplification and analysed the genotypes. Results: Modified Ziehl-Neelsen method revealed 5–20 Cryptosporidium oocysts detected per 10 Liter. Five out of the 55 (9.09%) surface water samples were found positive for Cryptosporidium spp. by Ziehl-Neelsen test and seven (12.7%) were found positive by nested PCR. The staining results were consistent with PCR. Seven Cryptosporidium PCR products were successfully sequenced and five gp60 subtypes were detected. Our finding of gp60 gene revealed that all of the positive isolates were Cryptosporidium parvum and belonged to subtype families IIa and IId. Conclusion: Our investigations were showed that collection of water samples were contaminated by Cryptosporidium, with potential hazards for the significant health problem. This study provides the first report on detection and genotyping of Cryptosporidium species from surface water samples in Iran, and its result confirmed the low clinical incidence of this parasite on the community.

Keywords: Cryptosporidium spp., membrane filtration, subtype, surface water, Iran

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Authors: Richard L. Summers

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The advanced computational analysis of biological systems is becoming increasingly dependent upon an understanding of the information-theoretic structure of the materials, energy and interactive processes that comprise those systems. The stability and survival of these living systems are fundamentally contingent upon their ability to acquire and process the meaning of information concerning the physical state of its biological continuum (biocontinuum). The drive for adaptive system reconciliation of a divergence from steady-state within this biocontinuum can be described by an information metric-based formulation of the process for actionable knowledge acquisition that incorporates the axiomatic inference of Kullback-Leibler information minimization driven by survival replicator dynamics. If the mathematical expression of this process is the Lagrangian integrand for any change within the biocontinuum then it can also be considered as an action functional for the living system. In the direct method of Lyapunov, such a summarizing mathematical formulation of global system behavior based on the driving forces of energy currents and constraints within the system can serve as a platform for the analysis of stability. As the system evolves in time in response to biocontinuum perturbations, the summarizing function then conveys information about its overall stability. This stability information portends survival and therefore has absolute existential meaning for the living system. The first derivative of the Lyapunov energy information function will have a negative trajectory toward a system's steady state if the driving force is dissipating. By contrast, system instability leading to system dissolution will have a positive trajectory. The direction and magnitude of the vector for the trajectory then serves as a quantifiable signature of the meaning associated with the living system’s stability information, homeostasis and survival potential.

Keywords: meaning, information, Lyapunov, living systems

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Authors: Yanhua Zhao, Yu Gou, Shu Feng, Dongdong Li, Chuanmin Tao

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The natural history of HBV infection could experience immune tolerant (IT), immune clearance (IC), HBeAg-negative inactive/quienscent carrier (ENQ), and HBeAg-negative hepatitis (ENH). As current biomarkers for discriminating these four phases have some weaknesses, additional serological indicators are needed. Hepatits B core-related antigen (HBcrAg) encoded with precore/core gene contains denatured HBeAg, HBV core antigen (HBcAg) and a 22KDa precore protein (p22cr), which was demonstrated to have a close association with natural history of hepatitis B infection, but no specific cutoff values and diagnostic parameters to evaluate the diagnostic efficacy. This study aimed to clarify the distribution of HBcrAg levels and evaluate its diagnostic performance during the natural history of infection from a Western Chinese perspective. 294 samples collected from treatment-naïve chronic hepatitis B (CHB) patients in different phases (IT=64; IC=72; ENQ=100, and ENH=58). We detected the HBcrAg values and analyzed the relationship between HBcrAg and HBV DNA. HBsAg and other clinical parameters were quantitatively tested. HBcrAg levels of four phases were 9.30 log U/mL, 8.80 log U/mL, 3.00 log U/mL, and 5.10 logU/mL, respectively (p < 0.0001). Receiver operating characteristic curve analysis demonstrated that the area under curves (AUCs) of HBcrAg and quantitative HBsAg at cutoff values of 9.25 log U/mL and 4.355 log IU/mL for distinguishing IT from IC phases were 0.704 and 0.694, with sensitivity 76.39% and 59.72%, specificity 53.13% and 79.69%, respectively. AUCs of HBcrAg and quantitative HBsAg at cutoff values of 4.15 log U/mlmL and 2.395 log IU/mlmL for discriminating between ENQ and ENH phases were 0.931 and 0.653, with sensitivity 87.93% and 84%, specificity 91.38% and 39%, respectively. Therefore, HBcrAg levels varied significantly among four natural phases of HBV infection. It had higher predictive performance than quantitative HBsAg for distinguishing between ENQ-patients and ENH-patients and similar performance with HBsAg for the discrimination between IT and IC phases, which indicated that HBcrAg could be a potential serological marker for CHB.

Keywords: chronic hepatitis B, hepatitis B core-related antigen, hepatitis B surface antigens, hepatitis B virus

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Authors: Mateen A. Khan, Elizabeth J. Theil, Dixie J. Goss

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Cellular iron homeostasis is accomplished by the coordinated regulated expression of iron uptake, storage, and export. Iron regulate the translation of ferritin and mitochondrial aconitase iron responsive element (IRE)-mRNA by interaction with an iron regulatory protein (IRPs). Iron increases protein biosynthesis encoded in iron responsive element. The noncoding structure IRE-mRNA, approximately 30-nt, folds into a stem loop to control synthesis of proteins in iron trafficking, cell cycling, and nervous system function. Fluorescence anisotropy measurements showed the presence of one binding site on IRP1 for ferritin and mitochondrial aconitase IRE-mRNA. Scatchard analysis revealed the binding affinity (Kₐ) and average binding sites (n) for ferritin and mitochondrial aconitase IRE-mRNA were 68.7 x 10⁶ M⁻¹ and 9.2 x 10⁶ M⁻¹, respectively. In order to understand the relative importance of equilibrium and stability, we further report the contribution of electrostatic interactions in the overall binding of two IRE-mRNA with IRP1. The fluorescence quenching of IRP1 protein was measured at different ionic strengths. The binding affinity of IRE-mRNA to IRP1 decreases with increasing ionic strength, but the number of binding sites was independent of ionic strength. Such results indicate a differential contribution of electrostatics to the interaction of IRE-mRNA with IRP1, possibly related to helix bending or stem interactions and an overall conformational change. Selective destabilization of ferritin and mitochondrial aconitase RNA/protein complexes as reported here explain in part the quantitative differences in signal response to iron in vivo and indicate possible new regulatory interactions.

Keywords: IRE-mRNA, IRP1, binding, ionic strength

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Authors: Ni̇sa Yeni̇kalayci, Türkan Aybi̇ke Akarca

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The ability of correlation the number and space relations, one of the basic scientific process skills, is being used in the transformation of a two-dimensional object into a three-dimensional image or in the expression of symmetry axes of the object. With this research, it is aimed to determine the ability of science students to establish number and space relations. The research was carried out with a total of 90 students studying in the first semester of the Science Education program of a state university located in the Turkey’s Black Sea Region in the fall semester of 2017-2018 academic year. An ‘Integrative Perception Test (IPT)’ was designed by the researchers to collect the data. Within the scope of IPT, the courses and workbooks specific to the field of science were scanned and the ones without symmetrical structure from the visual items belonging to the ‘Physics - Chemistry – Biology’ sub-fields were selected and listed. During the application, it was expected that students would imagine and draw images of the missing half of the visual items that were given incomplete in the first place. The data obtained from the test in which there are 30 images or pictures in total (f Physics = 10, f Chemistry = 10, f Biology = 10) were analyzed descriptively based on the drawings created by the students as ‘complete (2 points), incomplete/wrong (1 point), empty (0 point)’. For the teaching of new concepts in small aged groups, images or pictures showing symmetrical structures and similar applications can also be used.

Keywords: integrative perception, number and space relations, science education, scientific process skills

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Authors: Alexander Rych

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Piql Preservation Services (“Piql”) is a turnkey solution designed for secure, migration-free long- term preservation of digital data. Piql sets an open standard for long- term preservation for the future. It consists of equipment and processes needed for writing and retrieving digital data. Exponentially growing amounts of data demand for logistically effective and cost effective processes. Digital storage media (hard disks, magnetic tape) exhibit limited lifetime. Repetitive data migration to overcome rapid obsolescence of hardware and software bears accelerated risk of data loss, data corruption or even manipulation and adds significant repetitive costs for hardware and software investments. Piql stores any kind of data in its digital as well as analog form securely for 500 years. The medium that provides this is a film reel. Using photosensitive film polyester base, a very stable material that is known for its immutability over hundreds of years, secure and cost-effective long- term preservation can be provided. The film reel itself is stored in a packaging capable of protecting the optical storage medium. These components have undergone extensive testing to ensure longevity of up to 500 years. In addition to its durability, film is a true WORM (write once- read many) medium. It therefore is resistant to editing or manipulation. Being able to store any form of data onto the film makes Piql a superior solution for long-term preservation. Paper documents, images, video or audio sequences – all of those file formats and documents can be preserved in its native file structure. In order to restore the encoded digital data, only a film scanner, a digital camera or any appropriate optical reading device will be needed in the future. Every film reel includes an index section describing the data saved on the film. It also contains a content section carrying meta-data, enabling users in the future to rebuild software in order to read and decode the digital information.

Keywords: digital data, long-term preservation, migration-free, photosensitive film

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Authors: A. M. Pashayev, A. S. Samedov, T. B. Usubaliyev, N. Sh. Yusifov

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Different types of coating technologies are widely used for gas turbine blades. Thermal barrier coatings, consisting of ceramic top coat, thermally grown oxide and a metallic bond coat are used in applications for thermal protection of hot section components in gas turbine engines. Operational characteristics and longevity of high-temperature turbine blades substantially depend on a right choice of composition of the protective thermal barrier coatings. At a choice of composition of a coating and content of the basic elements it is necessary to consider following factors, as minimum distinctions of coefficients of thermal expansions of elements, level of working temperatures and composition of the oxidizing environment, defining the conditions for the formation of protective layers, intensity of diffusive processes and degradation speed of protective properties of elements, extent of influence on the fatigue durability of details during operation, using of elements with high characteristics of thermal stability and satisfactory resilience of gas corrosion, density, hardness, thermal conduction and other physical characteristics. Forecasting and a choice of a thermal barrier coating composition, all above factors at the same time cannot be considered, as some of these characteristics are defined by experimental studies. The implemented studies and investigations show that one of the main failures of coatings used on gas turbine blades is related to not fully taking the physical-chemical features of elements into consideration during the determination of the composition of alloys. It leads to the formation of more difficult spatial structure, composition which also changes chaotically in some interval of concentration that doesn't promote thermal and structural firmness of a coating. For the purpose of increasing the thermal and structural resistant of gas turbine blade coatings is offered a new approach to forecasting of composition on the basis of analysis of physical-chemical characteristics of alloys taking into account the size factor, electron configuration, type of crystal lattices and Darken-Gurry method. As a result, of calculations and experimental investigations is offered the new four-component metallic bond coat on the basis of chrome for the gas turbine blades.

Keywords: gas turbine blades, thermal barrier coating, metallic bond coat, strategic metals, physical-chemical features

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Authors: Carin Lindskog

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Introduction: Long-term success requires an organizational balance between continuity (exploitation) and change (exploration). The problem of balancing exploitation and exploration is a common issue in studies of organizational learning. In order to better face the tough competition in the face of changes, organizations need to exploit their current business and explore new business fields by developing new capabilities. The purpose of this work in progress is to develop a conceptual framework to shed light on the relevance of 'learning activities', i.e., exploitation and exploration, on different levels. The research questions that will be addressed are as follows: What sort of learning activities are found in the Business Process Management (BPM) field? How can these activities be linked to the individual level, group, level, and organizational level? In the work, a literature review will first be conducted. This review will explore the status of learning activities in the BPM field. An outcome from the literature review will be a conceptual framework of learning activities based on the included publications. The learning activities will be categorized to focus on the categories exploitation, exploration or both and into the levels of individual, group, and organization. The proposed conceptual framework will be a valuable tool for analyzing the research field as well as identification of future research directions. Related Work: BPM has increased in popularity as a way of working to strengthen the quality of the work and meet the demands of efficiency. Due to the increase in BPM popularity, more and more organizations reporting on BPM failure. One reason for this is the lack of knowledge about the extended scope of BPM to other business contexts that include, for example, more creative business fields. Yet another reason for the failures are the fact of the employees’ are resistant to changes. The learning process in an organization is an ongoing cycle of reflection and action and is a process that can be initiated, developed and practiced. Furthermore, organizational learning is multilevel; therefore the theory of organizational learning needs to consider the individual, the group, and the organization level. Learning happens over time and across levels, but it also creates a tension between incorporating new learning (feed-forward) and exploiting or using what has already been learned (feedback). Through feed-forward processes, new ideas and actions move from the individual to the group to the organization level. At the same time, what has already been learned feeds back from the organization to a group to an individual and has an impact on how people act and think.

Keywords: business process management, exploitation, exploration, learning activities

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Authors: Hafiza Javaria Ashraf, Xinghong Wang, Zhanghong Shi, Youming Hou

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Insect’s innate immunity depends on a variety of defense responses for the recognition of invading pathogens. Pathogen recognition involves particular proteins known as pattern recognition receptors (PRRs). These PRRs interact with pathogen-associated molecular patterns (PAMPs) present on the surface of pathogens to distinguish between self and non-self. C-type lectins (CTLs) belong to a superfamily of PPRs which involved in insect immunity and defense mechanism. Rhynchophorus ferrugineus Olivier is a devastating pest of Palm cultivations in China. Although studies on R. ferrugineus immune mechanism and host defense have conducted, however, the role of CTL in immune responses of R. ferrugineus remains elusive. Here, we report RfCTL, which is a secreted protein containing a single-CRD domain. The open reading frame (ORF) of CTL is 226 bp, which encodes a putative protein of 168 amino acids. Transcript expression analysis revealed that RfCTL highly expressed in immune-related tissues, i.e., hemolymph and fat body. The abundance of RfCTL in the gut and fat body dramatically increased upon Staphylococcus aureus and Escherichia coli bacterial challenges, suggesting a role in defense against gram-positive and gram-negative bacterial infection. Taken together, we inferred that RfCTL might be involved in the immune defense of R. ferrugineus and established a solid foundation for future studies on R. ferrugineus CTL domain proteins for better understanding of insect immunity.

Keywords: biological invasion, c-type lectin, insect immunity, Rhynchophorus ferrugineus Oliver

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Authors: Zeynep Tanriverdi̇

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Sustainable architecture is a design approach that provides healthy, comfortable, safe, clean space production as well as utilizes minimum resources for efficient and economical use of natural resources and energy. Traditional houses located in rural areas are sustainable structures built at the design and implementation stage in accordance with the climatic environmental data of the region and also effectively using natural energy resources. The fact that these structures are located in an earthquake geography like Türkiye brings their earthquake resistance to the agenda. Since the construction of these structures, which contain the architectural and technological cultural knowledge of the past, is shaped according to the characteristics of the regions where they are located, their resistance to earthquakes also differs. Analyses in rural areas after the earthquake show that there are light-damaged structures that can survive, severely damaged structures, and completely destroyed structures. In this regard, experts can implement repair, consolidation, and reconstruction applications, respectively. While simple repair interventions are carried out in accordance with the original data in traditional houses that have shown great resistance to earthquakes, reinforcement work blended with new technologies can be applied in damaged structures. In reconstruction work, a wide variety of applications can be seen with the possibilities of modern technologies. In rural areas experiencing earthquakes around the world, there are experimental new housing applications that are renewable, environmentally friendly, and sustainable with modern construction techniques in the light of scientific data. With these new residences, it is aimed to create earthquake-resistant, economical, healthy, and pain-relieving therapy spaces for people whose daily lives have been interrupted by disasters. In this study, the preservation of high earthquake-prone rural areas will be discussed through the knowledge transfer of traditional architecture and also permanent housing practices using new sustainable technologies to improve the area. In this way, it will be possible to keep losses to a minimum with sustainable, reliable applications prepared for the worst aspects of the disaster situation and to establish a link between the knowledge of the past and the new technologies of the future.

Keywords: sustainability, conservation, traditional construction systems and materials, new technologies, earthquake resistance

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Authors: Ga-Young Lee, Hyun-Man Kim

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The modulation of the cell-cell junction is critical in epithelial-mesenchymal transition during tumorigenesis. As RhoA activity is known to be up-regulated to dissociate cell-cell junction by contracting acto-myosin complex in various cancer cells, the present study investigated if RhoA activity was also associated with the disruption of the cell-cell junction of oral cancer cells. We studied SCC-25 cells which are established from oral squamous cell carcinoma if their E-cadherin junction (ECJ) was under control of RhoA. Interestingly, development of ECJ of SCC-25 cells depended on the amount of fibronectin (FN) coated on the culture dishes. Seeded cells promptly aggregated to develop ECJ on the substrates coated with a low amount of FN, whereas they were retarded in the development of ECJ on the substrates coated with a high amount of FN. However, it was an unexpected finding that total RhoA activity was lower in the dissociated cells on the substrates of high FN than in the aggregated cells on the substrates of low FN. Treating the dissociated cells on the substrates of high FN with LPA, a RhoA activator, promoted the development to ECJ. In contrast, treating the aggregated cells on the substrates of low FN with Clostridium botulinum C3, a toxin decreasing RhoA activity, dissociated cells concomitant with the disruption of ECJ. Genetical knockdown of RhoA expression by transfecting RhoA siRNA also down-regulated the development of ECJ in SCC-25 cells. Furthermore, PMA, an activator of protein kinase C (PKC), down-regulated the development of ECJ junction of SCC-25 cells on the substrates coated with low FN. In contrast, GO6976, a PKC inhibitor, up-regulated the development of ECJ of SCC-25 cells with the activation of RhoA on the substrates coated with high FN. In conclusion, in the present study, we demonstrated unexpected results that the activation of RhoA promotes the development of ECJ, whereas the inhibition of RhoA retards the development of ECJ in SCC-25 cells.

Keywords: E-cadherin junction, oral squamous cell carcinoma, PKC, RhoA, SCC-25

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Authors: Neslihan Demirci, Serdar Durdağı

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Mycobacterium tuberculosis is still a worldwide disease-causing agent that, according to WHO, led to the death of 1.5 million people from tuberculosis (TB) in 2020. The bacteria reside in macrophages located specifically in the lung. There is a known quadruple drug therapy regimen for TB consisting of isoniazid (INH), rifampin (RIF), pyrazinamide (PZA), and ethambutol (EMB). Over the past 60 years, there have been great contributions to treatment options, such as recently approved delamanid (OPC67683) and bedaquiline (TMC207/R207910), targeting mycolic acid and ATP synthesis, respectively. Also, there are natural compounds that can block the tryptophanyl-tRNA synthetase (TrpRS) enzyme, chuangxinmycin, and indolmycin. Yet, already the drug resistance is reported for those agents. In this study, the newly released TrpRS enzyme structure is investigated for potential inhibitor drugs from already synthesized molecules to help the treatment of resistant cases and to propose an alternative drug for the quadruple drug therapy of tuberculosis. Maestro, Schrodinger is used for docking and molecular dynamic simulations. In-house library containing ~8000 compounds among FDA-approved indole-containing compounds, a total of 57 obtained from the ChemBL were used for both ATP and tryptophan binding pocket docking. Best of indole-containing 57 compounds were subjected to hit expansion and compared later with virtual screening workflow (VSW) results. After docking, VSW was done. Glide-XP docking algorithm was chosen. When compared, VSW alone performed better than the hit expansion module. Best scored compounds were kept for ten ns molecular dynamic simulations by Desmond. Further, 100 ns molecular dynamic simulation was performed for elected molecules according to Z-score. The top three MMGBSA-scored compounds were subjected to steered molecular dynamic (SMD) simulations by Gromacs. While SMD simulations are still being conducted, ponesimod (for multiple sclerosis), vilanterol (β₂ adrenoreceptor agonist), and silodosin (for benign prostatic hyperplasia) were found to have a significant affinity for tuberculosis TrpRS, which is the propulsive force for the urge to expand the research with in vitro studies. Interestingly, top-scored ponesimod has been reported to have a side effect that makes the patient prone to upper respiratory tract infections.

Keywords: drug repurposing, molecular dynamics, tryptophanyl-tRNA synthetase, tuberculosis

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Authors: Stanislaw Kosmynka

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Although the genesis of the activity of fighting salafist radicalism in Spain dates back to the 80’s, the development of extremism of this kind manifested itself only in the next decade. Its first permanently functioning structures in this country in the second half of 90’s of 20th century came from Algieria and Syria. At the same time it should be emphasized that this distinction is in many dimensions conventional, the more so because they consisted also of immigrants from other coutries of Islam, particularly from Morocco. The paper seeks to understand the radical salafist challenge for Spain in the context of some terrorist networks consisted of immigrants from Morocco. On the eve of the new millennium Moroccan jihadists played an increasingly important role. Although the activity of these groups had for many years mainly logistical and propaganda character, the bomb attack carried out on 11 March 2004 in Madrid constituted an expression of open forms of terrorism, directed against the authorities and society of Spain and reflected the narration of representatives of the trend of the global jihad. The people involved in carrying out that act of violence were to a large extent Moroccan immigrants; also in the following years among the cells of radicals in Spain Moroccans stood out many times. That is why the forms and directions of activity of these extremists in Spain, also after 11th March 2004 and in the actual context of the impact of Islamic State, are worth presenting. The paper is focused on threats to the security of Spain and the region and remains connected with the issues of mutual relations of the society of a host country with immigrant communities which to a large degree come from this part of Maghreb.

Keywords: jihadi terrorism, Morocco, radical salafism, security, Spain, terrorist cells, threat

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Authors: Debit Datta, Jeffrey J. Coleman, Scott A. Enebak, Lori G. Eckhardt

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Loblolly pine (Pinus taeda) is a leading productive timber species in the southeastern USA. Over the past three years, an emerging threat is expressed by successive needle defoliation followed by stunted growth and tree mortality in loblolly pine plantations. Considering economic significance, it has now become a rising concern among landowners, forest managers, and forest health state cooperators. However, the symptoms of the disease were perplexed somewhat with root disease(s) and recurrently attributed to invasive Phytophthora species due to the similarity of disease nature and devastation. Therefore, the study investigated the potential causal agent of this disease and characterized the fungi associated with loblolly pine needle defoliation in the southeastern USA. Besides, 70 trees were selected at seven long-term monitoring plots at Chatom, Alabama, to monitor and record the annual disease incidence and severity. Based on colony morphology and ITS-rDNA sequence data, a total of 28 species of fungi representing 17 families have been recovered from diseased loblolly pine needles. The native brown-spot pathogen, Lecanosticta acicola, was the species most frequently recovered from unhealthy loblolly pine needles in combination with some other common needle cast and rust pathogen(s). Identification was confirmed using morphological similarity and amplification of translation elongation factor 1-alpha gene region of interest. Tagged trees were consistently found chlorotic and defoliated from 2019 to 2020. The current emergence of the brown-spot pathogen causing loblolly pine mortality necessitates the investigation of the role of changing climatic conditions, which might be associated with increased pathogen pressure to loblolly pines in the southeastern USA.

Keywords: brown-spot needle blight, loblolly pine, needle defoliation, plantation forestry

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Authors: Navodit Goel, Prabir K. Paul

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Tomato (Solanum lycopersicum) is attacked by Pseudomonas syringae pv. tomato causing speck lesions on the leaves leading to severe economic casualty. In the present study, aqueous fruit extracts of Azadirachta indica (neem) were sprayed on a single node of tomato plants grown under controlled contamination-free conditions. The treatment of plants was performed with neem fruit extract either alone or along with the pathogen. The parameters of observation were activities of polyphenol oxidase (PPO) and lysozyme, and isoform analysis of PPO; both at the treated leaves as well as untreated leaves away from the site of extract application. Polyphenol oxidase initiates phenylpropanoid pathway resulting in the synthesis of quinines from cytoplasmic phenols and production of reactive oxygen species toxic to broad spectrum microbes. Lysozyme is responsible for the breakdown of bacterial cell wall. The results indicate the upregulation of PPO and lysozyme activities in both the treated and untreated leaves along with de novo expression of newer PPO isoenzymes (which were absent in control samples). The appearance of additional PPO isoenzymes in bioelicitor-treated plants indicates that either the isoenzymes were expressed after bioelicitor application or the already expressed but inactive isoenzymes were activated by it. Lysozyme activity was significantly increased in the plants when treated with the bioelicitor or the pathogen alone. However, no new isoenzymes of lysozyme were expressed upon application of the extract. Induction of resistance by neem fruit extract could be a potent weapon in eco-friendly plant protection strategies.

Keywords: Azadirachta indica, lysozyme, polyphenol oxidase, Solanum lycopersicum

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Authors: Seyedfateh Moradi, Abas Ali Rahbar

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The present study focuses on investigating group psychoanalysis in the Middle East. The study uses a descriptive-analytic method and library resources have been used to collect the data. Additionally, the researcher’s observations of people’s everyday behavior have played an important role in the production and analysis of the study. Group psychoanalysis in the Middle East can be conducted through people’s daily behaviors, proverbs, poetry, mythology, etc., and some of the general characteristics of people in the Middle East include: xenophobia, revivalism, fatalism, nostalgic, wills and so on. Members of the group have often failed to achieve Libido wills and it is very important in unifying and reproduction violence. Therefore, if libidinal wills are irrationally fixed, it will be important in forming fundamentalist and racist groups, a situation that is dominant among many groups in the Middle East. Adversities, from early childhood and afterwards, in the subjects have always been influential in the political behavior of group members, and it manifests itself as counter-projections. Consequently, it affects the foreign policy of the governments. On the other hand, two kinds of subjects are identifiable in the Middle East, one; classical subject that is related to nostalgia and mythology and, two; modern subjects which is self-alienated. As a result, both subjects are seeking identity and self-expression in public in relation to forming groups. Therefore, collective unconscious in the Middle East shows itself as extreme boundaries and leads to forming groups characterized with violence. Psychoanalysis shows important aspects to identify many developments in the Middle East; totally analysis of Freud, Carl Jung and Reich about groups can be applied in the present Middle East.

Keywords: political, psychoanalysis, group, Middle East

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Authors: Sabrina Rahman

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The Mosque is Islam’s most symbolic structure, as well as the expression of collective identity. From the explicit words of our Prophet, 'The earth has been created for me as a masjid and a place of purity, and whatever man from my Ummah finds himself in need of prayer, let him pray' (anywhere)! it is obvious that a devout Muslim does not require a defined space or structure for divine worship since the whole earth is his prayer house. Yet we see that from time immemorial man throughout the Muslim world has painstakingly erected innumerable mosques. However, mosque design spans time, crosses boundaries, and expresses cultures. It is a cultural manifestation as much as one based on a regional building tradition or a certain interpretation of religion. The trend to express physical signs of religion is not new. Physical forms seem to convey symbolic messages. However, in recent times physical forms of mosque architecture are dominantly demising from mosque architecture projects in Bangladesh. Dome & minaret, the most prominent symbol of the mosque, is replacing by contextual and contemporary improvisation rather than subcontinental mosque architecture practice of early fellows. Thus the recent mosque projects of the last 15 years established the contemporary architectural realm in their design. Contextually, spiritual lighting, the serenity of space, tranquility of outdoor spaces, the texture of materials is widely establishing a new genre of Muslim prayer space. A case study based research will lead to specify its significant factors of modernism. Based on the findings, the paper presents evidence of recent projects as well as a guideline for the future image of contemporary Mosque architecture in Bangladesh.

Keywords: contemporary architecture, modernism, prayer space, symbolism

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Authors: Kristina Sakalauskiene, Renaldas Raisutis, Gintare Linkeviciute, Skaidra Valiukeviciene

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Cutaneous melanoma is a melanocytic skin tumour, which has a very poor prognosis while is highly resistant to treatment and tends to metastasize. Thickness of melanoma is one of the most important biomarker for stage of disease, prognosis and surgery planning. In this study, we hypothesized that the automatic analysis of spectrophotometric images and high-frequency ultrasonic 2D data can improve differential diagnosis of cutaneous melanoma and provide additional information about tumour penetration depth. This paper presents the novel complex automatic system for non-invasive melanocytic skin tumour differential diagnosis and penetration depth evaluation. The system is composed of region of interest segmentation in spectrophotometric images and high-frequency ultrasound data, quantitative parameter evaluation, informative feature extraction and classification with linear regression classifier. The segmentation of melanocytic skin tumour region in ultrasound image is based on parametric integrated backscattering coefficient calculation. The segmentation of optical image is based on Otsu thresholding. In total 29 quantitative tissue characterization parameters were evaluated by using ultrasound data (11 acoustical, 4 shape and 15 textural parameters) and 55 quantitative features of dermatoscopic and spectrophotometric images (using total melanin, dermal melanin, blood and collagen SIAgraphs acquired using spectrophotometric imaging device SIAscope). In total 102 melanocytic skin lesions (including 43 cutaneous melanomas) were examined by using SIAscope and ultrasound system with 22 MHz center frequency single element transducer. The diagnosis and Breslow thickness (pT) of each MST were evaluated during routine histological examination after excision and used as a reference. The results of this study have shown that automatic analysis of spectrophotometric and high frequency ultrasound data can improve non-invasive classification accuracy of early-stage cutaneous melanoma and provide supplementary information about tumour penetration depth.

Keywords: cutaneous melanoma, differential diagnosis, high-frequency ultrasound, melanocytic skin tumours, spectrophotometric imaging

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Authors: O. Davies, K. Veravalli, P. Panwalkar, M. Tofighi, P. Butterick, B. Healy, A. Mofidi

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Vancomycin resistant enterococcus infection is a condition that usually impacts ICUs, transplant, dialysis, and cancer units, often as a nosocomial infection. After an outbreak in the acute trauma and orthopaedic unit in Morriston hospital, we aimed to access the conditions that predispose VRE infections in our unit. Thirteen cases of VRE infection and five cases of VRE colonisations were identified in patients who were treated for orthopaedic care between 1/1/2020 and 1/11/2021. Cases were reviewed to identify predisposing factors, specifically looking at age, presenting condition and treatment, presence of infection and antibiotic care, active haemo-oncological condition, long term renal dialysis, previous hospitalisation, VRE predisposition, and clearance (PREVENT) scores, and outcome of care. The presenting condition, treatment, presence of postoperative infection, VRE scores, age was compared between colonised and the infected cohort. VRE type in both colonised and infection group was Enterococcus Faecium in all but one patient. The colonised group had the same age (T=0.6 P>0.05) and sex (2=0.115, p=0.74), presenting condition and treatment which consisted of peri-femoral fixation or arthroplasty in all patients. The infected group had one case of myelodysplasia and four cases of chronic renal failure requiring dialysis. All of the infected patient had sustained an infected complication of their fracture fixation or arthroplasty requiring reoperation and antibiotics. The infected group had an average VRE predisposition score of 8.5 versus the score of 3 in the colonised group (F=36, p<0.001). PREVENT score was 7 in the infected group and 2 in the colonised group(F=153, p<0.001). Six patients(55%) succumbed to their infection, and one VRE infection resulted in limb loss. In the orthopaedic cohort, VRE infection is a nosocomial condition that has peri-femoral predilection and is seen in association with immunosuppression or renal failure. The VRE infection cohort has been treated for infective complication of original surgery weeks prior to VRE infection. Based on our findings, we advise avoidance of infective complications, change of practice in use of antibiotics and use radical surgery and surveillance for VRE infections beyond infective precautions. PREVENT score shows that the infected group are unlikely to clear their VRE in the future but not the colonised group.

Keywords: surgical site infection, enterococcus, orthopaedic surgery, vancomycin resistance

Procedia PDF Downloads 122

Authors: K. Cartelier, D. Aime, V. Vernoud, J. Buitink, J. M. Prosperi, K. Gallardo, C. Le Signor

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Legumes can produce protein-rich seeds without nitrogen fertilizer through root symbiosis with nitrogen-fixing rhizobia. Rich in lysine, these proteins are used for human nutrition and animal feed. However, the instability of seed protein yield and quality due to environmental fluctuations limits the wider use of legumes such as pea. Breeding efforts are needed to optimize and stabilize seed nutritional value, which requires to identify the genetic determinism of seed protein plasticity in response to the environment. Towards this goal, we have studied the plasticity of protein content and composition of seeds from a collection of 200 Medicago truncatula ecotypes grown under four controlled conditions (optimal, drought, and winter/spring sowing). A quantitative analysis of one-dimensional protein profiles of these mature seeds was performed and plasticity indices were calculated from each abundant protein band. Genome-Wide Association Studies (GWAS) from these data identified major GWAS hotspots, from which a list of candidate genes was obtained. A Gene Ontology Enrichment Analysis revealed an over-representation of genes involved in several amino acid metabolic pathways. This led us to propose that environmental variations are likely to modulate amino acid balance, thus impacting seed protein composition. The selection of candidate genes for controlling the plasticity of seed protein composition was refined using transcriptomics data from developing Medicago truncatula seeds. The pea orthologs of key genes were identified for functional studies by mean of TILLING (Targeting Induced Local Lesions in Genomes) lines in this crop. We will present how this study highlighted mechanisms that could govern seed protein plasticity, providing new cues towards the stabilization of legume seed quality.

Keywords: GWAS, Medicago truncatula, plasticity, seed, storage proteins

Procedia PDF Downloads 127

Authors: Siddharth Deshpande, Nihar Masurkar, Vallerinteavide Mavelli Girish, Malan Desai, Chester Drum

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Folding and stabilization of recombinant proteins remain a consistent challenge for industrial and therapeutic applications. Proteins derived from thermophilic bacteria often have superior expression and stability qualities. To develop a generalizable approach to protein folding and stabilization, we tested the hypothesis that wrapping a thermostable exoshell around a protein substrate would aid folding and impart thermostable qualities to the internalized substrate. To test the effect of internalizing a protein within a thermostable exoshell (tES), we tested in vitro folding and stability using green fluorescent protein (GFPuv), horseradish peroxidase (HRP) and renilla luciferase (rLuc). The 8nm interior volume of a thermostable ferritin assembly was engineered to accommodate foreign proteins and either present a positive, neutral or negative interior charge environment. We further engineered the tES complex to reversibly assemble and disassemble with pH titration. Template proteins were expressed as inclusion bodies and an in vitro folding protocol was developed that forced proteins to fold inside a single tES. Functional yield was improved 100-fold, 100-fold and 150-fold with use of tES for GFPuv, HRP and rLuc respectively and was highly dependent on the internal charge environment of the tES. After folding, functional proteins could be released from the tES folding cavity using size exclusion chromatography at pH 5.8. Internalized proteins were tested for improved stability against thermal, organic, urea and guanidine denaturation. Our results demonstrated that thermostable exoshells can efficiently refold and stabilize inactive aggregates into functional proteins.

Keywords: thermostable shell, in vitro folding, stability, functional yield

Procedia PDF Downloads 229

Authors: Muge Unal, Ahmet Cilek

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Urban identity, having a dynamic characteristic spatial and semantic aspects, is a phenomenon in an ever-changing. Urban identity formation includes not only a process of physical nature but also development and change processes that take place in the political, economic, social and cultural values, whether national and international level. Although the concept of urban transformation is basically regarded as the spatial transformation; in fact, it reveals a holistic perspective and transformation based on dialectical relationship existing between the spatial and social relationship. For this reason, urban renewal needs to address as not only spatial but also the impact of spatial transformation on social, cultural and economic. Implementation tools used in the perception of urban transformation are varied concepts such as urban renewal, urban resettlement, urban rehabilitation, urban redevelopment, and urban revitalization. The phenomenon of urban transformation begins with the Industrial Revolution. Until the 1980s, it was interpreted as reconsidering physical fossil on urban environment factor like occurring in rapid urbanization, changing in the spatial structure of the city, concentrating of the population in urban areas. However, after the 1980s, it has resided in a conceptual structure which requires to be addressed physical, economic, social, technological and integrity of information. In conclusion, urban transformation, when it enter the literature as a practice of planning, has been up to date in terms of the conceptual structure and content and also hasn’t remained behind converting itself. Urban transformation still maintains its simplest expression, while it transforms so fast converts the contents. In this study, the relationship between urban design and components of urban transformation were discussed with strategies used as a place in the historical process of urban transformation besides a general evaluation of the concept of urban renewal.

Keywords: conceptual approach, urban identity, urban regeneration, urban renewal

Procedia PDF Downloads 409

Authors: Mahesh M. Patil, K. A. Anu-Appaiah

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Overweight and obesity is a serious medical problem, increasing in prevalence, and affecting millions worldwide. Investigators have been trying from decades to articulate the burden of obesity and related risk factors. To answer this problem, we suggest a new therapeutic anti-obesity compounds from Garcinia xanthochymus fruit. However, there is little published scientific information on non-hydroxycitric acid Garcinia species. Our findings include biochemical characterization of the fruit; in vivo toxicity and bio-efficacy study of G. xanthochymus in high fat diet wistar rat model. We observed that Garcinia pericarp is a rich source of organic acids, polyphenols, mono- (40.63%) and poly-unsaturated fatty acids (16.45%; omega-3: 10.02%). Toxicological studies have showed that Garcinia is safe and had no observed adverse effect level up to 400 mg/kg/day. Body weight and food intake was significantly (P<0.05) reduced in oral gavage treated rats (sonicated Garcinia powder) in 13 weeks. Subcutaneous fat was significantly (P<0.05) reduced in Garcinia treated rats. Hepatocytes significantly (p<0.05) overexpressed sterol regulatory element binding protein 2, liver X receptor- α, liver X receptor- β, lipoprotein lipase and monoacylglycerol lipase. Fatty acid binding protein 1 and peroxisome proliferator activated receptor- α were down regulated as assessed by real time qPCR. Currently our research is focused on the adipocyte obesity related gene expressions, effect of Garcinia on 3T3-adipocyte cell lines and high fat diet induced mice model. This in vivo pre-clinical data suggests that G. xanthochymus may have clinical utility for the treatment of obesity. However, further studies are required to establish its potency.

Keywords: Garcinia xanthochymus, anti-obesity, high fat diet, real time qPCR

Procedia PDF Downloads 242

Authors: Daniel Canales, Fabián Alvarez, Pablo Varela, Marcela Saavedra, Claudio García, Paula Zapata

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Calcium oxide nanoparticles (n-CaO) ca. 8 nm were obtained from eggshell waste. The n-CaO was incorporated into Poly(lactic acid) PLA matrix in 10 and 20 wt.% of filler content by electrospinning process to obtain PLA/n-CaO nanocomposite fibers as a potential use in scaffold for bone tissue regeneration. The fibers morphology and diameter were homogeneity, the PLA had a diameter of 2.2 ± 0.8 µm and, with the nanoparticles incorporation (20wt.%), reached ca. 2.9 ± 0.9 µm. The PLA/n-CaO nanocomposites fibers showed in vitro bioactivity, capable of inducing the precipitation of hydroxyapatite (HA) layer in the fiber surface after 7 days in Simulated Body Solution (SBF). The biocidal and biological properties of PLA/n-Cao with 20 wt.% were evaluated, showing a 30% reduction in bacterial viability against S. aureus and 11% for E. coli after 6 hours of bacterial suspensions exposure. Furthermore, the fibers did not show a cytotoxic effect on the bone marrow ST-2 cell line, permitting the cell adhesion and proliferation in Roswell Park Memorial Institute medium (RPMI). The PLA/n-CaO with 20 wt.% of nanoparticles showed a higher capacity to promote the osteogenic differentiation, significantly increasing the alkaline phosphatase (ALP) expression after 7 days compared to PLA and cell control. The in vivo analysis corroborated the biocompatibility of scaffolds prepared, the presence of n-CaO in PLA reduced the formation of fibrous encapsulation of the material improve the healing process.

Keywords: electrospun scaffolds, PLA based nanocomposites, calcium oxide nanoparticles, bioactive materials, tissue engineering

Procedia PDF Downloads 74

Authors: Rohit Kothari, Muneer Mohamed, Vivekanandh K., Sunmeet Sandhu, Preema Sinha, Anuj Bhatnagar

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Introduction: Hyper-IgE syndrome is a rare primary immunodeficiency syndrome characterised by triad of severe atopic dermatitis, recurrent pulmonary infections, and recurrent staphylococcal skin infections. The diagnosis requires a high degree of suspicion, typical clinical features, and not mere rise in serum-IgE levels, which may be seen in multiple conditions. Genetic studies are not always possible in a resource poor setting. This study highlights various presentations of Hyper-IgE syndrome in skin of color children. Case-series: Our study had six children of Hyper-IgE syndrome aged twomonths to tenyears. All had onset in first ten months of life except one with a late-onset at two years. All had recurrent eczematoid rash, which responded poorly to conventional treatment, secondary infection, multiple episodes of hospitalisation for pulmonary infection, and raised serum IgE levels. One case had occasional vesicles, bullae, and crusted plaques over both the extremities. Genetic study was possible in only one of them who was found to have pathogenic homozygous deletions of exon-15 to 18 in DOCK8 gene following which he underwent bone marrow transplant (BMT), however, succumbed to lower respiratory tract infection two months after BMT and rest of them received multiple courses of antibiotics, oral/ topical steroids, and cyclosporine intermittently with variable response. Discussion: Our study highlights various characteristics, presentation, and management of this rare syndrome in children. Knowledge of these manifestations in skin of color will facilitate early identification and contribute to optimal care of the patients as representative data on the same is limited in literature.

Keywords: absolute eosinophil count, atopic dermatitis, eczematous rash, hyper-immunoglobulin E syndrome, pulmonary infection, serum IgE, skin of color

Procedia PDF Downloads 121

Authors: Yan Li, Qingxiang Meng, Bo Zhou, Zhenming Zhou

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The objective of this study was to compare the effects of ensiled mulberry leaves (EML), and sun-dried mulberry fruit pomace (SMFP) on fecal bacterial communities in Simmental crossbred finishing steers fed the following 3 diets: a standard TMR diet, standard diet containing EML and standard diet containing SMFP, and the diets had similar protein and energy levels. Bacterial communities in the fecal content were analyzed using Illumina Miseq sequencing of the V4 region of the 16S rRNA gene amplification. Quantitative real-time PCR was used to detect the selected bacterial species in the feces. Most of the sequences were assigned to phyla Firmicutes (56.67%) and Bacteroidetes(35.90%), followed by Proteobacteria(1.86%), Verrucomicrobia(1.80%) and Tenericutes(1.37%). And the predominant genera included the 5-7N15 (5.91%), CF231 (2.49%), Oscillospira (2.33%), Paludibacter (1.23%) and Akkermansia(1.11%). As for the treatments, no significant differences were observed in Firmicutes (p = 0.28), Bacteroidetes (p = 0.63), Proteobacteria (p = 0.46), Verrucomicrobia (p = 0.17) and Tenericutes (p = 0.75). On the genus level, classified genera with high abundance (more than 0.1%) mainly came from two phyla: Bacteroidetes and Firmicutes. Also no differences were observed in most genera level, 5-7N15 (p = 0.21), CF231 (p = 0.62), Oscillospira (p = 0.9), Paludibacter (p = 0.33) and Akkermansia (p = 0.37), except that rc4-4 were lower in the CON and SMFP groups compared to the EML animals (p = 0.02). Additionally, there were no differences in richness estimate and diversity indices (p > 0.16), and treatments had no significant effect on most selected bacterial species in the fecal (p > 0.06), except that Ruminococcus albus were higher in the EML group (p < 0.01) and Streptococcus bovis were lower in the CON group (p < 0.01). In conclusion, diets supplemented with EML and SMFP have little influence on fecal bacterial community composition in finishing steers.

Keywords: fecal bacteria community composition, sequencing, ensiled mulberry leaves (EML), sun-dried mulberry fruit pomace (SMFP)

Procedia PDF Downloads 301

Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim

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Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate and characterize strains of Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from thirty one chicken cecal and fecal samples collected from a local farm. In vitro studies were performed to assess antibacterial activity (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), susceptibility to antibiotics (amoxycillin, cotrimoxazole, chloramphenicol, cefuroxime, ceftriaxone, ciprofloxacin, and nalidixic acid), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified using biochemical and molecular assays (API 50 CHL, and API ZYM kits followed by 16S rDNA gene sequence analysis). Two strains were identified, of which, Enteroccocus faecium was capable of inhibiting the growth of S. enteritidis and was susceptible to a wide range of antibiotics. In addition, the isolated strain exhibited significant resistance under highly acidic conditions (pH=2.5) for 8 hours and survived well in bile salt at 0.2% for 24 hours and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, the E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: acid tolerance, antimicrobial activity, Enterococcus faecium, probiotic

Procedia PDF Downloads 387

Authors: Xing Yan Fan, Xing Lin Xu, Man Yuan Chen, Pei Tzu Lee, Yu Ting Wang, Yi Xiao Cao, Rui Yao

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Pyramid sales have been a widespread issue in China. Victims who are defrauded not only lose money but damage interpersonal relationship. A deeper understanding of pyramid-sale models can be beneficial to prevent potential victims from fraud and improve the property security. The goals of this study were to detect psychological characteristics of pyramid-sale sellers, and analyse persuasive skills in pyramid organizations. A qualitative study was conducted in this study. Participants (n=6) recruited by 'snowball' sampling from present pyramid-sale sellers (n=3) and imprisoned pyramid-sale sellers (n=3). All participants accepted semi-structured interview for collecting data. Content analysis was adopted for data coding and analysis. The results indicate that pyramid organizations are used to utilize their appearance packaging and celebrity effect to strengthen the positions in participants’ mind. The status gap between pyramid-sale sellers in same organization, as well as rewards to increase reputation, are used to motivate participants in pyramid. The most significant common characteristics among all participants are that they tend to possess a high sense of belongingness within the firm. Moreover, the expression of pyramid-sale sellers on gambling mentality is expected to growth as constantly losing money. Findings suggest that the psychological characteristics of pyramid-sale sellers in accordance with Maslow’s hierarchy of needs, persuasive skills of pyramid organization confront to 'attitude-behaviour change model'. These findings have implication on 'immune education' that providing guidance for victims out of stuck and protecting ordinary people from the jeopardizing of pyramid sales.

Keywords: pyramid sales, characteristics, persuasive skills, qualitative study

Procedia PDF Downloads 240

Authors: Ananya Govindarajan

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Tetrahymena thermophila is a single-cell, eukaryotic organism that belongs to the Protozoa Kingdom. Tetrahymena thermophila is often used in signal transduction pathway studies because of its ability to model sensory input and the effects of environmental conditions such as chemicals and temperature. The recently discovered G37 chemorepellent receptor showed increased responsiveness to all chemorepellents. Investigating the mutant G37 Tetrahymena gene in various test solutions, including ferric chloride, ferrous sulfate, hydrogen peroxide, tetrazolium blue, potassium chloride, and dithiothreitol were performed to determine the role of oxidants and reducing agents with the mutant and wild-type cells (CU427) to assess the role of the receptor. Behavioral assays and recordings processed by ImageJ indicated that ferric chloride, hydrogen peroxide, and tetrazolium blue yielded little to no chemorepellent responses from G37 cells (<20% ARs). CU427 cells were over-responsive based on the mean percent of cells (>50% ARs). Reducing agents elicited chemorepellent responses from both G37 and CU427, in addition to potassium chloride. Cell responses were classified as over-responsive (>50% ARs). Dithiothreitol yielded unexpected results as G37 (37.0% ARs) and CU427 (38.1% ARs) had relatively similar responses and were only responsive and not over-responsive to the reducing agent test chemical solution. Ultimately, this indicates that the G37 receptor is more interactive with molecules that are reducing agents or non-oxidant compounds; G37 may be unable to sense and respond to oxidants effectively, further elucidating the pathways of the G37 strain and nature of this receptor. Results also indicate that the CSF most likely contained an oxidant, like ferric chloride. This research can be further applied to neuronal influences and how specific compounds may affect human neurons individually and their excitability as the responses model action potentials and membrane potential.

Keywords: tetrahymena thermophila, signal transduction, chemosensory, oxidant, reducing agent

Procedia PDF Downloads 118

Authors: Ridwaan N. Milase, Leonardo J. Van Zyl, Marla Trindade

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American foulbrood (AFB) is the world’s most devastating honeybee disease that has drastically reduced the population of Apis mellifera capensis since 2009. The outbreak has jeopardized the South African bee keeping industry as well as the agricultural sector dependent on honeybees for honey production and pollination, leading to significant economic losses. AFB is caused by Paenibacillus larvae, a spore-forming, Gram positive facultative anaerobic and flagellated bacterium. The use of antibiotics within beehives has selected for resistant strains of P. larvae, while the current practice of burning spore contaminated beehives and equipment contributes to the economic losses in the honeybee-keeping industry. Therefore, phage therapy is proposed as a promising alternative to combat P. larvae strains affecting A. mellifera capensis. The genomes of two P. larvae strains isolated from infected combs in the Western Cape have been sequenced and annotated using bioinformatics tools. Genome analyses has revealed that these P. larvae strains are lysogens to more than 6 different prophages and possess different type of clustered regularly interspaced short palindromic repeat (CRISPRs) regions per strain. Active prophages from one of the two P. larvae strains were detected and identified using PCR. Electron microscopy was used to determine the family of the identified active prophages. Lytic bacteriophages that specifically target the two P. larvae strains were purified from sewage wastewater, beehive materials, and soil samples to investigate their potential development as anti-P. larvae agents. Another alternative treatment being investigated is the development of a prophage endolysin cocktail. Endolysin genes of the prophages have been targeted, cloned and expressed in Escherichia coli. The heterologously expressed endolysins have been purified and are currently being assessed for their lytic activity against P. larvae strains and other commensal microorganisms that compose the honeybee larvae microbiota. The study has shown that phage therapy and endolysins have a great potential as alternative control methods for AFB disease affecting A. mellifera capensis.

Keywords: American foulbrood, bacteriophage, honeybee, Paenibacillus larvae

Procedia PDF Downloads 165