Search results for: Isolation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 888

Search results for: Isolation

348 Food Security of Migrants in a Regional Area of Australia: A Qualitative Study

Authors: Joanne Sin Wei Yeoh, Quynh Lê, Rosa McManamey

Abstract:

Food security indicates the ability of individuals, households and communities to acquire food that is healthy, sustainable, affordable, appropriate and accessible. Despite Australia’s current ability to produce enough food to feed a population larger than its current population, there has been substantial evidence over the last decades to demonstrate many Australians struggle to feed themselves, including those from a cultural and linguistically diverse (CALD) background. The study aimed to investigate migrants’ perceptions and experiences on food security in Tasmania. Semi-structured interviews were conducted with 33 migrants residing in North, South and North West Tasmania, who were recruited through purposive sampling. Thematic analysis was employed to analyse the interview data. Four main themes were identified from the interview data: (1) Understanding of food security; (2) Experiences with the food security in Tasmania; (3) Factors that influence migrants’ food security in Tasmania; and (4) Acculturation strategies. Various sub-themes have emerged under each of these four major themes. Though the findings indicate participants are satisfied with their current food security in Tasmania, they still encounter some challenges in food availability, accessibility, and affordability in Tasmania. Factors that influence migrants’ food security were educational background, language barrier, socioeconomic status, geographical isolation, and cultural background. By using different acculturation strategies, migrants managed to adapt to the new food culture. In addition, social and cultural capitals were also treated as vital roles in improving migrants’ food security. The findings indicate migrants residing in Tasmania face different challenges on food security. They use different strategies for food security while acculturating into a new environment. The findings may provide useful information for migrants in Australia and various private organisations or relevant government departments that address food security for migrants.

Keywords: experiences, food security, migrants, perceptions

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347 Exploring Challenges Faced by People Living with HIV/AIDS After Disclosure in Sub-Saharan Countries

Authors: Veliswa Nonfundo Hoho, Jabulani Gilford Kheswa

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HIV/AIDS has been a long-term condition worldwide, which does not only affect physical health but also causes psychological and social challenges in people living with this condition. In Sub-Saharan countries, namely; Nigeria, Uganda, Zimbabwe and South Africa, people living with HIV/AIDS come across different challenges especially after one has disclosed his/her status. They experience stigma and discrimination, isolation, lack of accessing and receiving treatment, lack of support and experience psychological distress. By using the evidence-based systematic review as a form of methodology, journal articles, dissertations, internet, and books were explored. This paper seeks to describe the challenges faced by people living with HIV/AIDS after disclosure, which forms a critical component of HIV/AIDS prevention and treatment interventions. The disclosure process model is used to underpin the study. This theory allows one to understand when and why interpersonal and verbal self-disclosure is beneficial for individuals who live with concealable stigmatized identities such as HIV/AIDS. Literature findings advocate that both negative and positive results were noted after disclosing one’s HIV status and psychosocial well-being of the majority of people living with HIV/AIDS also get affected especially in societies which subscribe HIV/AIDS pandemic to witchcraft. As for the infected homosexuals, research indicates that they suffer in silence and to cover their emotional emptiness due to ostracism, they often report low- self-efficacy with regard to condom use and become susceptible to reinfections which further place their lives at heightened risk for low immune system. In this regard, this paper challenges the policies which protect the dignity of people living with HIV/AIDS and calls for unity and financial support in favour of psychoeducational programmes and support groups aimed at curbing discrimination.

Keywords: disclosure, discrimination, homosexuality, self-efficacy

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346 Anti-Tyrosinase and Antibacterial Activities of Marine Fungal Extracts

Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

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A variety of genetic and environmental factors cause various cosmetics and dermatological problems. There are already claimed drugs available in market for treating these problems. However, the challenge remains in finding more potent, environmental friendly, causing minimal side effects and economical cosmeceuticals. This leads to an increased demand for natural cosmeceutical products in the last few decades. Plant derived ingredients are limited because plants either contain toxic metabolites, grow too slow or seasonal harvesting is a problem. The research work carried out in this project aims at isolation, characterization of marine fungal secondary metabolite and evaluating their potential use in future cosmetic skin care products. We have isolated and purified 35 morphologically different fungal isolates from various marine habitats of the India. These isolates have been functionally characterized for anti-tyrosinase, antioxidant and anti-acne activities. For molecular characterization, the Internal Transcribed spacer (ITS) region of 15 functionally active marine fungal isolates was amplified using universal primers, ITS1 and ITS4 and sequenced. Out of 15 marine fungal isolates crude extract of strains D4 (Aspergillus terreus) and P2 (Talaromyces stipitatus) showed 70% and 57% tyrosinase inhibition at 1mg/mL respectively. Strain D5 (Simplicillium lamellicola) has showed significant inhibition against Propionibacterium acnes and Staphylococcus epidermidis. In addition, all these strains also displayed DPPH- radical scavenging activity and may be utilized as skin cosmeceutical applications. Purification and characterization of crude extracts for identification of active lead molecule is under process.

Keywords: anti-acne, anti-tyrosinase, cosmeceutical, marine fungi

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345 Theory of the Optimum Signal Approximation Clarifying the Importance in the Recognition of Parallel World and Application to Secure Signal Communication with Feedback

Authors: Takuro Kida, Yuichi Kida

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In this paper, it is shown a base of the new trend of algorithm mathematically that treats a historical reason of continuous discrimination in the world as well as its solution by introducing new concepts of parallel world that includes an invisible set of errors as its companion. With respect to a matrix operator-filter bank that the matrix operator-analysis-filter bank H and the matrix operator-sampling-filter bank S are given, firstly, we introduce the detail algorithm to derive the optimum matrix operator-synthesis-filter bank Z that minimizes all the worst-case measures of the matrix operator-error-signals E(ω) = F(ω) − Y(ω) between the matrix operator-input-signals F(ω) and the matrix operator-output-signals Y(ω) of the matrix operator-filter bank at the same time. Further, feedback is introduced to the above approximation theory, and it is indicated that introducing conversations with feedback do not superior automatically to the accumulation of existing knowledge of signal prediction. Secondly, the concept of category in the field of mathematics is applied to the above optimum signal approximation and is indicated that the category-based approximation theory is applied to the set-theoretic consideration of the recognition of humans. Based on this discussion, it is shown naturally why the narrow perception that tends to create isolation shows an apparent advantage in the short term and, often, why such narrow thinking becomes intimate with discriminatory action in a human group. Throughout these considerations, it is presented that, in order to abolish easy and intimate discriminatory behavior, it is important to create a parallel world of conception where we share the set of invisible error signals, including the words and the consciousness of both worlds.

Keywords: matrix filterbank, optimum signal approximation, category theory, simultaneous minimization

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344 Transformer Life Enhancement Using Dynamic Switching of Second Harmonic Feature in IEDs

Authors: K. N. Dinesh Babu, P. K. Gargava

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Energization of a transformer results in sudden flow of current which is an effect of core magnetization. This current will be dominated by the presence of second harmonic, which in turn is used to segregate fault and inrush current, thus guaranteeing proper operation of the relay. This additional security in the relay sometimes obstructs or delays differential protection in a specific scenario, when the 2nd harmonic content was present during a genuine fault. This kind of scenario can result in isolation of the transformer by Buchholz and pressure release valve (PRV) protection, which is acted when fault creates more damage in transformer. Such delays involve a huge impact on the insulation failure, and chances of repairing or rectifying fault of problem at site become very dismal. Sometimes this delay can cause fire in the transformer, and this situation becomes havoc for a sub-station. Such occurrences have been observed in field also when differential relay operation was delayed by 10-15 ms by second harmonic blocking in some specific conditions. These incidences have led to the need for an alternative solution to eradicate such unwarranted delay in operation in future. Modern numerical relay, called as intelligent electronic device (IED), is embedded with advanced protection features which permit higher flexibility and better provisions for tuning of protection logic and settings. Such flexibility in transformer protection IEDs, enables incorporation of alternative methods such as dynamic switching of second harmonic feature for blocking the differential protection with additional security. The analysis and precautionary measures carried out in this case, have been simulated and discussed in this paper to ensure that similar solutions can be adopted to inhibit analogous issues in future.

Keywords: differential protection, intelligent electronic device (IED), 2nd harmonic inhibit, inrush inhibit

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343 Phylogenetic Differential Separation of Environmental Samples

Authors: Amber C. W. Vandepoele, Michael A. Marciano

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Biological analyses frequently focus on single organisms, however many times, the biological sample consists of more than the target organism; for example, human microbiome research targets bacterial DNA, yet most samples consist largely of human DNA. Therefore, there would be an advantage to removing these contaminating organisms. Conversely, some analyses focus on a single organism but would greatly benefit from the additional information regarding the other organismal components of the sample. Forensic analysis is one such example, wherein most forensic casework, human DNA is targeted; however, it typically exists in complex non-pristine sample substrates such as soil or unclean surfaces. These complex samples are commonly comprised of not just human tissue but also microbial and plant life, where these organisms may help gain more forensically relevant information about a specific location or interaction. This project aims to optimize a ‘phylogenetic’ differential extraction method that will separate mammalian, bacterial and plant cells in a mixed sample. This is accomplished through the use of size exclusion separation, whereby the different cell types are separated through multiple filtrations using 5 μm filters. The components are then lysed via differential enzymatic sensitivities among the cells and extracted with minimal contribution from the preceding component. This extraction method will then allow complex DNA samples to be more easily interpreted through non-targeting sequencing since the data will not be skewed toward the smaller and usually more numerous bacterial DNAs. This research project has demonstrated that this ‘phylogenetic’ differential extraction method successfully separated the epithelial and bacterial cells from each other with minimal cell loss. We will take this one step further, showing that when adding the plant cells into the mixture, they will be separated and extracted from the sample. Research is ongoing, and results are pending.

Keywords: DNA isolation, geolocation, non-human, phylogenetic separation

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342 Inhibitory Activity of Podospermum canum and Its Active Components on Collagenase, Elastase and Hyaluronidase Enzymes

Authors: Ozlem Bahadir Acikara, Mert Ilhan, Ekin Kurtul, Karel Smejkal, Esra Kupeli Akkol

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Present study is aimed to investigate in vitro inhibitory effects of the extracts prepared from the aerial parts of Podospermum canum (Asteraceae) on hyaluronidase, collagenase, and elastase enzymes using a bioassay-guided fractionation. Inhibitory effects of the extract, sub-extracts, fractions obtained by column chromatography, and isolated compounds on collagenase, elastase, and hyaluronidase were performed by using in vitro enzyme inhibitory assays based on spectrophotometric evaluation. The ethyl acetate and remaining water extracts prepared from the plant displayed significant inhibitory activities on collagenase and elastase, while petroleum ether and chloroform extracts did not show any inhibitory activity. Eleven known compounds: arbutin, 6'-O-caffeoylarbutin, cichoriin, 3,5-dicaffeoylquinic acid methyl ester, apigenin-7-O-β-glucoside, luteolin-7-O-β-glucoside, apigenin-7-O-β-rutinoside, isoorientin, orientin, vitexin, procatechuic acid, and compound 4-hydroxy-benzoic acid 4-(6-O-α-rhamnopyranosyl-β-glucopyranosyl) benzyl ester have been obtained from ethyl acetate sub-extract of the plant through bioassay-guided fractionation and isolation. Results of the present study have revealed that among the isolated compounds, apigenin-7-O-β-glucoside, luteolin-7-O-β-glucoside, apigenin-7-O-β-rutinoside and isoorientin showed potent enzyme inhibitory activities. However, methanolic extract of P. canum displayed a greater inhibitory activity than fractions and isolated compounds both on collagenase and elastase.

Keywords: Asteraceae, collagenase, elastase, hyaluronidase, Podospermum canum

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341 Natural Gas Flow Optimization Using Pressure Profiling and Isolation Techniques

Authors: Syed Tahir Shah, Fazal Muhammad, Syed Kashif Shah, Maleeha Gul

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In recent days, natural gas has become a relatively clean and quality source of energy, which is recovered from deep wells by expensive drilling activities. The recovered substance is purified by processing in multiple stages to remove the unwanted/containments like dust, dirt, crude oil and other particles. Mostly, gas utilities are concerned with essential objectives of quantity/quality of natural gas delivery, financial outcome and safe natural gas volumetric inventory in the transmission gas pipeline. Gas quantity and quality are primarily related to standards / advanced metering procedures in processing units/transmission systems, and the financial outcome is defined by purchasing and selling gas also the operational cost of the transmission pipeline. SNGPL (Sui Northern Gas Pipelines Limited) Pakistan has a wide range of diameters of natural gas transmission pipelines network of over 9125 km. This research results in answer a few of the issues in accuracy/metering procedures via multiple advanced gadgets for gas flow attributes after being utilized in the transmission system and research. The effects of good pressure management in transmission gas pipeline network in contemplation to boost the gas volume deposited in the existing network and finally curbing gas losses UFG (Unaccounted for gas) for financial benefits. Furthermore, depending on the results and their observation, it is directed to enhance the maximum allowable working/operating pressure (MAOP) of the system to 1235 PSIG from the current round about 900 PSIG, such that the capacity of the network could be entirely utilized. In gross, the results depict that the current model is very efficient and provides excellent results in the minimum possible time.

Keywords: natural gas, pipeline network, UFG, transmission pack, AGA

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340 Impact of Social Distancing on the Correlation Between Adults’ Participation in Learning and Acceptance of Technology

Authors: Liu Yi Hui

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The COVID-19 pandemic in 2020 has globally affected all aspects of life, with social distancing and quarantine orders causing turmoil and learning in community colleges being temporarily paused. In fact, this is the first time that adult education has faced such a severe challenge. It forces researchers to reflect on the impact of pandemics on adult education and ways to respond. Distance learning appears to be one of the pedagogical tools capable of dealing with interpersonal isolation and social distancing caused by the pandemic. This research aims to examine whether the impact of social distancing during COVID-19 will lead to increased acceptance of technology and, subsequently, an increase in adults ’ willingness to participate in distance learning. The hypothesis that social distancing and the desire to participate in distance learning affects learners’ tendency to accept technology is investigated. Teachers ’ participation in distance education and acceptance of technology are used as adjustment variables with the relationship to “social distancing,” “participation in distance learning,” and “acceptance of technology” of learners. A questionnaire survey was conducted over a period of twelve months for teachers and learners at all community colleges in Taiwan who enrolled in a basic unit course. Community colleges were separated using multi-stage cluster sampling, with their locations being metropolitan, non-urban, south, and east as criteria. Using the G*power software, 660 samples were selected and analyzed. The results show that through appropriate pedagogical strategies or teachers ’ own acceptance of technology, adult learners’ willingness to participate in distance learning could be influenced. A diverse model of participation can be developed, improving adult education institutions’ ability to plan curricula to be flexible to avoid the risk associated with epidemic diseases.

Keywords: social distancing, adult learning, community colleges, technology acceptance model

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339 Improving the Exploitation of Fluid in Elastomeric Polymeric Isolator

Authors: Haithem Elderrat, Huw Davies, Emmanuel Brousseau

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Elastomeric polymer foam has been used widely in the automotive industry, especially for isolating unwanted vibrations. Such material is able to absorb unwanted vibration due to its combination of elastic and viscous properties. However, the ‘creep effect’, poor stress distribution and susceptibility to high temperatures are the main disadvantages of such a system. In this study, improvements in the performance of elastomeric foam as a vibration isolator were investigated using the concept of Foam Filled Fluid (FFFluid). In FFFluid devices, the foam takes the form of capsule shapes, and is mixed with viscous fluid, while the mixture is contained in a closed vessel. When the FFFluid isolator is affected by vibrations, energy is absorbed, due to the elastic strain of the foam. As the foam is compressed, there is also movement of the fluid, which contributes to further energy absorption as the fluid shears. Also, and dependent on the design adopted, the packaging could also attenuate vibration through energy absorption via friction and/or elastic strain. The present study focuses on the advantages of the FFFluid concept over the dry polymeric foam in the role of vibration isolation. This comparative study between the performance of dry foam and the FFFluid was made according to experimental procedures. The paper concludes by evaluating the performance of the FFFluid isolator in the suspension system of a light vehicle. One outcome of this research is that the FFFluid may preferable over elastomer isolators in certain applications, as it enables a reduction in the effects of high temperatures and of ‘creep effects’, thereby increasing the reliability and load distribution. The stiffness coefficient of the system has increased about 60% by using an FFFluid sample. The technology represented by the FFFluid is therefore considered by this research suitable for application in the suspension system of a light vehicle.

Keywords: FFFluid, dry foam, anti-vibration devices, elastomeric polymer foam

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338 Cas9-Assisted Direct Cloning and Refactoring of a Silent Biosynthetic Gene Cluster

Authors: Peng Hou

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Natural products produced from marine bacteria serve as an immense reservoir for anti-infective drugs and therapeutic agents. Nowadays, heterologous expression of gene clusters of interests has been widely adopted as an effective strategy for natural product discovery. Briefly, the heterologous expression flowchart would be: biosynthetic gene cluster identification, pathway construction and expression, and product detection. However, gene cluster capture using traditional Transformation-associated recombination (TAR) protocol is low-efficient (0.5% positive colony rate). To make things worse, most of these putative new natural products are only predicted by bioinformatics analysis such as antiSMASH, and their corresponding natural products biosynthetic pathways are either not expressed or expressed at very low levels under laboratory conditions. Those setbacks have inspired us to focus on seeking new technologies to efficiently edit and refractor of biosynthetic gene clusters. Recently, two cutting-edge techniques have attracted our attention - the CRISPR-Cas9 and Gibson Assembly. By now, we have tried to pretreat Brevibacillus laterosporus strain genomic DNA with CRISPR-Cas9 nucleases that specifically generated breaks near the gene cluster of interest. This trial resulted in an increase in the efficiency of gene cluster capture (9%). Moreover, using Gibson Assembly by adding/deleting certain operon and tailoring enzymes regardless of end compatibility, the silent construct (~80kb) has been successfully refactored into an active one, yielded a series of analogs expected. With the appearances of the novel molecular tools, we are confident to believe that development of a high throughput mature pipeline for DNA assembly, transformation, product isolation and identification would no longer be a daydream for marine natural product discovery.

Keywords: biosynthesis, CRISPR-Cas9, DNA assembly, refactor, TAR cloning

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337 Puerto Rico and Pittsburg: A Social Psychology Perspective on How Perceived Infringement on Job and Cultural Identity Unite Racially Different Working-Class Groups

Authors: Reagan Rodriguez

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With a growing divide between political echo chambers in the United States, exacerbated by race and income inequality, it might seem to be unfathomable to draw connections that tie working class in an industrial city and a U.S. territory. Yet, in regions where either the economy has been hit due to dwindling job infrastructure or natural disasters have left indelible marks on an island already once marked by colonial imperialism, a larger social shared identity is at play. Fracking has long been an intergenerational and stable work opportunity for many in the Pittsburg PA, yet the rising severity of global climate change may soon impact the policy and even presidential elections which could result in the reduction of jobs in the industry. Cock-fighting, considered a cultural mainstay within the island of Puerto Rico, has already had legislation banning activity and thus cutting out one of the most lucrative aspects of a severely injured economy. Insecurity, infringement, and isolation while being tied to a working-class bracket with no other opportunities in proximity have left both groups expressing similar frustration and while another larger shared identity politic is giving little other options to develop social mobility. This paper utilizes a thematic analysis and compares convergent and divergent themes on internet forums amongst unionized fracking workers in Pittsburg and cockfighters in Puerto Rico. This research examines how group identity in relation to job and cultural identity is most strong and at which points its most malleable; when intergenerational job identity becomes a part of one’s cultural identity, its override may be strongest when it is perceived as threatened. Final findings and limitations were comprehensively outlined.

Keywords: identity threat, social psychology, group identity, culture and social mobility

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336 Illness Experience Without Illness: A Qualitative Study on the Lived Experience of Young Adults During the COVID-19 Pandemic

Authors: Gemma Postil, Claire Zanin, Michael Halpin, Caroline Ritter

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Illness experience research typically focuses on people that are living with a medical condition; however, the broad consequences of the COVID-19 pandemic are impacting those without the virus itself, as many experienced extensive lockdowns, social isolation, and distress. Drawing on conceptual work in the illness experience literature, we argue that policy and social changes tied to COVID-19 produce biographical disruptions. In this sense, we argue that the COVID-19 pandemic produces illness experience without illness, as the pandemic comprehensively impacts health and biography. This paper draws on 30 in-depth interviews with young adults living in Prince Edward Island (PEI), which were conducted as part of a larger project to understand how young adults navigate compliance with the COVID-19 pandemic. We then inductively analyzed the interviews with a constructivist grounded theory approach. Specifically, we demonstrate that young adults living in PEI during the COVID-19 pandemic experienced biographical disruptions throughout the pandemic despite not contracting the virus. First, we detail how some participants experience biographical acceleration, with the pandemic accelerating relationships, home buying, and career planning. Second, we demonstrate biographical stagnation, wherein participants report being unable to pursue major life milestones. Lastly, we describe biographical regression, wherein participants feel they are losing ground during the pandemic and are actively falling behind their peers. These findings provide the novel application of illness experience concepts to the context of the COVID-19 pandemic, contribute to work on illness experience and ambiguity, and extend Bury’s conceptualization of biographical disruption. In conclusion, we demonstrate that young adults experienced the biographical disruption expected from having COVID-19 without having an illness, highlighting the depth to which the pandemic affected young adults.

Keywords: illness experience, lived experience, biographical disruption, COVID-19, young adults

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335 Applications of Forensics/DNA Tools in Combating Gender-Based Violence: A Case Study in Nigeria

Authors: Edeaghe Ehikhamenor, Jennifer Nnamdi

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Introduction: Gender-based violence (GBV) was a well-known global crisis before the COVID-19 pandemic. The pandemic burden only intensified the crisis. With prevailing lockdowns, increased poverty due to high unemployment, especially affecting females, and other mobility restrictions that have left many women trapped with their abusers, plus isolation from social contact and support networks, GBV cases spiraled out of control. Prevalence of economic with cultural disparity, which is greatly manifested in Nigeria, is a major contributory factor to GBV. This is made worst by religious adherents where the females are virtually relegated to the background. Our societal approaches to investigations and sanctions to culprits have not sufficiently applied forensic/DNA tools in combating these major vices. Violence against women or some rare cases against men can prevent them from carrying out their duties regardless of the position they hold. Objective: The main objective of this research is to highlight the origin of GBV, the victims, types, contributing factors, and the applications of forensics/DNA tools and remedies so as to minimize GBV in our society. Methods: Descriptive information was obtained through the search on our daily newspapers, electronic media, google scholar websites, other authors' observations and personal experiences, plus anecdotal reports. Results: Findings from our exploratory searches revealed a high incidence of GBV with very limited or no applications of Forensics/DNA tools as an intervening mechanism to reduce GBV in Nigeria. Conclusion: Nigeria needs to develop clear-cut policies on forensics/DNA tools in terms of institutional framework to develop a curriculum for the training of all stakeholders to fast-track justice for victims of GBV so as to serve as a deterrent to other culprits.

Keywords: gender-based violence, forensics, DNA, justice

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334 Metagenomics-Based Molecular Epidemiology of Viral Diseases

Authors: Vyacheslav Furtak, Merja Roivainen, Olga Mirochnichenko, Majid Laassri, Bella Bidzhieva, Tatiana Zagorodnyaya, Vladimir Chizhikov, Konstantin Chumakov

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Molecular epidemiology and environmental surveillance are parts of a rational strategy to control infectious diseases. They have been widely used in the worldwide campaign to eradicate poliomyelitis, which otherwise would be complicated by the inability to rapidly respond to outbreaks and determine sources of the infection. The conventional scheme involves isolation of viruses from patients and the environment, followed by their identification by nucleotide sequences analysis to determine phylogenetic relationships. This is a tedious and time-consuming process that yields definitive results when it may be too late to implement countermeasures. Because of the difficulty of high-throughput full-genome sequencing, most such studies are conducted by sequencing only capsid genes or their parts. Therefore the important information about the contribution of other parts of the genome and inter- and intra-species recombination to viral evolution is not captured. Here we propose a new approach based on the rapid concentration of sewage samples with tangential flow filtration followed by deep sequencing and reconstruction of nucleotide sequences of viruses present in the samples. The entire nucleic acids content of each sample is sequenced, thus preserving in digital format the complete spectrum of viruses. A set of rapid algorithms was developed to separate deep sequence reads into discrete populations corresponding to each virus and assemble them into full-length consensus contigs, as well as to generate a complete profile of sequence heterogeneities in each of them. This provides an effective approach to study molecular epidemiology and evolution of natural viral populations.

Keywords: poliovirus, eradication, environmental surveillance, laboratory diagnosis

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333 Using Baculovirus Expression Vector System to Express Envelop Proteins of Chikungunya Virus in Insect Cells and Mammalian Cells

Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu

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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

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332 From Being to Becoming: Emancipation and Empowerment in the African Diaspora

Authors: R. Vidhya

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Diasporic writings present a comprehensive view of social, cultural and psychological dualities of immigrants. Isolation and the strong feelings of insecurity and inferiority due to constant marginalization coupled with a nostalgia for their motherland, its customs, culture, language, food and people which keep haunting the minds of immigrants are the major themes that are handled by diasporic writers. In the African diaspora, more than the men, it is the women who face the brunt and burden of the triple jeopardy – the racial, class and gender discrimination. Women writers from Africa have successfully sketched the plight of African women in the diaspora. Buchi Emecheta, a Nigerian woman writer deftly portrays the African Diaspora in her novels. She skillfully weaves her stories with her own experiences as an immigrant in the United Kingdom. She portrays the immigrant life and psychology through numerous themes like exile, geographical shift of locations, transactions of culture, political instability and the dilemma of moral and religious ideologies in her diasporic novels Second-class Citizen, Gwendolyn and Kehinde. The contemporary Nigerian woman writer Chimamanda Ngozi Adichie has also dexterously depicted the diasporic dilemma of her protagonist Ifemelu in Americannah, who initially has the experience of a despondent and a downcast in the United States of America. This paper aims to analyse the diasporic sentiments and sensibilities of the Nigerian Igbo women writers Buchi Emecheta and Chimamanda Ngozi Adichie whose women characters finally find emancipation and empowerment in the African Diaspora. This study is based on the Africana Womanist Literary theory propounded by Clenora Hudson-Weems.

Keywords: African Diaspora, Nigerian women writers, Buchi Emecheta, Chimamanda Ngozi Adichie, emancipation, empowerment

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331 Identitarian Speech in Exile by Representatives of Central Europe

Authors: Georgiana Ciobotaru

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The experience of exile is a defining one for the mittleeuropean writers, which is also the generator of an identity discourse manifested in the plan of fiction. In exile, the authors often build their marginality in opposition to that deserted mundi center. The Polish Gombrowicz carried out his existence, for more than twenty-three years, in a geographical exile, distancing himself from his country, and, from a cultural point of view, the writing meant a possibility of escape, of plunge into a literary exile that often constituted a way of conditioning the practice of writers. He opted for one of the attitudes that a writer in exile may have, namely he preferred to continue speaking Polish, although he was far from his homeland, turning to the public in his homeland, his entire literary creation in exile being promoted through Kulturia, the Paris-based immigration magazine. The problem of exile must be constantly related to three essential aspects, namely: territory, identity and language. The exile, both the writer and his characters, displays a characteristic attitude towards the abandoned land, but also towards the adoptive, towards the mother tongue, but also towards the idiom encountered, thus proving an original manner in terms of how it asserts, de-builds or re-builds its identity. In these texts written after leaving Poland, a series of open works by Trans-Atlantic, Gombrowicz assumes and internalizes the inadequacy between his self and the reality outside to make it the principle of his perception of the world. The expression of marginality that characterized the texts developed when the writer was still in Poland seems to acquire a certain coherence against the background of a logic imposed on the new experience, namely that of exile. Texts created during his exile in Argentina appear in a different context, in other words, in a situation of inadequacy towards the world: ignorance of the language, poverty, isolation that characterizes especially the first years spent there. This study aims to highlight how the Polish author de-builds and reconstructs his Mittel-European identity profile through language.

Keywords: discourse, exile, identity, immigration

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330 Preventive Effect of Three Kinds of Bacteriophages to Control Vibrio coralliilyticus Infection in Oyster Larvae

Authors: Hyoun Joong Kim, Jin Woo Jun, Sib Sankar Giri, Cheng Chi, Saekil Yun, Sang Guen Kim, Sang Wha Kim, Jeong Woo Kang, Se Jin Han, Se Chang Park

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Vibrio corallilyticus is a well-known pathogen of coral. It is also infectious to a variety of shellfish species, including Pacific oyster (Crassostrea gigas) larvae. V. corallilyticus is remained to be a major constraint in marine bivalve aquaculture practice, especially in artificial seed production facility. Owing to the high mortality and contagious nature of the pathogen, large amount of antibiotics has been used for disease prevention and control. However, indiscriminate use of antibiotics may result in food and environmental pollution, and development of antibiotic resistant strains. Therefore, eco-friendly disease preventative measures are imperative for sustainable bivalve culture. The present investigation proposes the application of bacteriophage (phage) as an effective alternative method for controlling V. corallilyticus infection in marine bivalve hatcheries. Isolation of phages from sea water sample was carried out using drop or double layer agar methods. The host range, stability and morphology of the phage isolates were studied. In vivo phage efficacy to prevent V. corallilyticus infection in oyster larvae was also performed. The isolated phages, named pVco-5 and pVco-7 was classified as a podoviridae and pVco-14, was classified as a siphoviridae. Each phages were infective to four strains of seven V. corallilyticus strains tested. When oyster larvae were pre-treated with the phage before bacterial challenge, mortality of the treated oyster larvae was lower than that in the untreated control. This result suggests that each phages have the potential to be used as therapeutic agent for controlling V. corallilyticus infection in marine bivalve hatchery.

Keywords: bacteriophage, Vibrio coralliilyticus, Oyster larvae, mortality

Procedia PDF Downloads 199
329 Testing of Complicated Bus Bar Protection Using Smart Testing Methodology

Authors: K. N. Dinesh Babu

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In this paper, the protection of a complicated bus arrangement with a dual bus coupler and bus sectionalizer using low impedance differential protection applicable for very high voltages like 220kV and 400kV is discussed. In many power generation stations, several operational procedures are implemented to utilize the transfer bus as the main bus and to facilitate the maintenance of circuit breakers and current transformers (in each section) without shutting down the bay(s). Owing to this fact, the complications in operational philosophy have thrown challenges for the bus bar protection implementation. Many bus topologies allow any one of the main buses available in the station to be used as an auxiliary bus. In such a system, pre-defined precautions and procedures are made as guidelines, which are followed before assigning any bus as an auxiliary bus. The procedure involves shifting of links, changing rotary switches, insertion of test block, and so on, thereby causing unreliable operation. This kind of unreliable operation or inadvertent procedural lapse may result in the isolation of the bus bar from the grid due to the unpredictable operation of the bus bar protection relay, which is a commonly occurring phenomenon due to manual mistakes. With the sophisticated configuration and implementation of logic in modern intelligent electronic devices, the operator is free to select the transfer arrangement without sacrificing the protection required by a bus differential system for a reliable operation, and labor-intensive processes are completely eliminated. This paper deals with the procedure to test the security logic for such special scenarios using Megger make SMRT, bus bar protection relay to assure system stability and get rid of all the specific operational precautions/procedure.

Keywords: bus bar protection, by-pass isolator, blind spot, breaker failure, intelligent electronic device, end fault, bus unification, directional principle, zones of protection, breaker re-trip, under voltage security, smart megger relay tester

Procedia PDF Downloads 49
328 Accessing Single Parenting and Disabled Children: A Case Study of Ghana

Authors: Edwina Owusu Panin

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Families may face significant obstacles as a result of single parenting and disabilities. The amenities and support those single parents need to give their children with disabilities the care they need are frequently out of their reach. These can include financial hardship, limited access to health and education, and social isolation. In addition, cultural attitudes toward disability can worsen these challenges, making it difficult for families to get the support and resources they need. Despite these challenges, many single parents have shown resilience and strength to overcome these difficulties and defend the rights of their children; some, too, have failed in taking care of their disabled children in Ghana. The study traces the developmental process of how single parents cope with disabled children. There is a discouraging fact that single father’s face a much more dreadful task in taking care of their disabled children in Ghana, which is later highlighted in the article. Additional research and support are needed to address the unique needs of families facing these challenges. This case study explores the experiences of single parents raising children with disabilities in Ghana. Using a qualitative approach, the study examines the challenges facing lone parents in caring for children, including access to healthcare, education and social support. In addition, the study examines the impact of cultural disability attitudes on the experiences of single parents and their children and what causes it in Ghana. Findings indicate that single parents in Ghana face significant challenges in accessing resources and support for their children and that cultural attitudes toward disability may aggravate these challenges. However, the study recommends the tenacity and strengths of how to create awareness, protect the welfare and also by encouraging single parents to face these challenges and protect the rights of their children, swaying away influences of bad cultural attitudes.

Keywords: disability, single parenting, case study, assessing

Procedia PDF Downloads 57
327 In vivo Genotoxicity Testing of Sesbania Grandiflora (Katuray) Flower Methanolic Extract

Authors: Levylee Bautista, Dawn Grace Santos, Aishwarya Veluchamy, Jesusa Santos, Ghafoor, Jr. I Haque, Rodolfo Rafael

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The booming interest in using natural compounds as an alternative to conventional medications has paved way to focus the attention on plants that provide rich sources of bioactive phytochemicals. For regulatory purposes, evaluation of the genotoxic effects of such alternatives is therefore empirical as part of the plant’s hazard assessment. Sesbania grandiflora is among the plants used as a traditional remedy in folk medicine and a subject of research for its medicinal benefits. This study aimed to evaluate the genotoxic potential induced by S. grandiflora flower methanol extract (SGFME) in terms of the frequency of micronucleus (MN) in polychromatic erythrocyte (PCE) (MNPCE) and PCE ratio employing the micronucleus assay. The frequency of MN was examined in bone marrow cells (BMCs) obtained from male Swiss albino mice exposed in vivo to four different concentrations (11.25, 22.5, 40, and 90 mg/kg) of SGFME and MMC (70 mg/kg; positive control) and sacrificed 24 hours post-intraperitoneal injection. Results showed a significant (p < 0.01) rate of MNPCEs for 11.25 and 22.5 tested concentrations of SGFME and is comparable with the MMC-treated mice. Although PCE ratio values in all doses of SGFME-treated mice were over 0.20, it is worth noting that 40 and 90 tested concentrations of SGFME-treated mice exhibited the lowest value, i.e., 0.22 and 0.28, respectively. The present study has demonstrated that S. grandiflora possesses genotoxic potential for murine BMCs. Such activity could be ascribed from the bioactive compounds present in S. grandiflora that require further isolation and characterization of the active molecules. Likewise, findings of this study warrant a caution of the use of S. grandiflora insomuch as further investigations do not demonstrate their safety.

Keywords: genotoxicity, micronucleus, phytochemicals, Sesbania grandiflora

Procedia PDF Downloads 117
326 Category-Base Theory of the Optimum Signal Approximation Clarifying the Importance of Parallel Worlds in the Recognition of Human and Application to Secure Signal Communication with Feedback

Authors: Takuro Kida, Yuichi Kida

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We show a base of the new trend of algorithm mathematically that treats a historical reason of continuous discrimination in the world as well as its solution by introducing new concepts of parallel world that includes an invisible set of errors as its companion. With respect to a matrix operator-filter bank that the matrix operator-analysis-filter bank H and the matrix operator-sampling-filter bank S are given, firstly, we introduce the detailed algorithm to derive the optimum matrix operator-synthesis-filter bank Z that minimizes all the worst-case measures of the matrix operator-error-signals E(ω) = F(ω) − Y(ω) between the matrix operator-input-signals F(ω) and the matrix operator-output signals Y(ω) of the matrix operator-filter bank at the same time. Further, feedback is introduced to the above approximation theory and it is indicated that introducing conversations with feedback does not superior automatically to the accumulation of existing knowledge of signal prediction. Secondly, the concept of category in the field of mathematics is applied to the above optimum signal approximation and is indicated that the category-based approximation theory is applied to the set-theoretic consideration of the recognition of humans. Based on this discussion, it is shown naturally why the narrow perception that tends to create isolation shows an apparent advantage in the short term and, often, why such narrow thinking becomes intimate with discriminatory action in a human group. Throughout these considerations, it is presented that, in order to abolish easy and intimate discriminatory behavior, it is important to create a parallel world of conception where we share the set of invisible error signals, including the words and the consciousness of both worlds.

Keywords: signal prediction, pseudo inverse matrix, artificial intelligence, conditional optimization

Procedia PDF Downloads 133
325 Isolation and Characterisation of Novel Environmental Bacteriophages Which Target the Escherichia coli Lamb Outer Membrane Protein

Authors: Ziyue Zeng

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Bacteriophages are viruses which infect bacteria specifically. Over the past decades, phage λ has been extensively studied, especially its interaction with the Escherichia coli LamB (EcLamB) protein receptor. Nonetheless, despite the enormous numbers and near-ubiquity of environmental phages, aside from phage λ, there is a paucity of information on other phages which target EcLamB as a receptor. In this study, to answer the question of whether there are other EcLamB-targeting phages in the natural environment, a simple and convenient method was developed and used for isolating environmental phages which target a particular surface structure of a particular bacterium; in this case, the EcLamB outer membrane protein. From the enrichments with the engineered bacterial hosts, a collection of EcLamB-targeting phages (ΦZZ phages) were easily isolated. Intriguingly, unlike phage λ, an obligate EcLamB-dependent phage in the Siphoviridae family, the newly isolated ΦZZ phages alternatively recognised EcLamB or E. coli OmpC (EcOmpC) as a receptor when infecting E. coli. Furthermore, ΦZZ phages were suggested to represent new species in the Tequatrovirus genus in the Myoviridae family, based on phage morphology and genomic sequences. Most phages are thought to have a narrow host range due to their exquisite specificity in receptor recognition. With the ability to optionally recognise two receptors, ΦZZ phages were considered relatively promiscuous. Via the heterologous expression of EcLamB on the bacterial cell surface, the host range of ΦZZ phages was further extended to three different enterobacterial genera. Besides, an interesting selection of evolved phage mutants with a broader host range was isolated, and the key mutations involved in their evolution to adapt to new hosts were investigated by genomic analysis. Finally, and importantly, two ΦZZ phages were found to be putative generalised transducers, which could be exploited as tools for DNA manipulations.

Keywords: environmental microbiology, phage, microbe-host interactions, microbial ecology

Procedia PDF Downloads 72
324 Deproteinization of Moroccan Sardine (Sardina pilchardus) Scales: A Pilot-Scale Study

Authors: F. Bellali, M. Kharroubi, Y. Rady, N. Bourhim

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In Morocco, fish processing industry is an important source income for a large amount of by-products including skins, bones, heads, guts, and scales. Those underutilized resources particularly scales contain a large amount of proteins and calcium. Sardina plichardus scales from resulting from the transformation operation have the potential to be used as raw material for the collagen production. Taking into account this strong expectation of the regional fish industry, scales sardine upgrading is well justified. In addition, political and societal demands for sustainability and environment-friendly industrial production systems, coupled with the depletion of fish resources, drive this trend forward. Therefore, fish scale used as a potential source to isolate collagen has a wide large of applications in food, cosmetic, and biomedical industry. The main aim of this study is to isolate and characterize the acid solubilize collagen from sardine fish scale, Sardina pilchardus. Experimental design methodology was adopted in collagen processing for extracting optimization. The first stage of this work is to investigate the optimization conditions of the sardine scale deproteinization on using response surface methodology (RSM). The second part focus on the demineralization with HCl solution or EDTA. And the last one is to establish the optimum condition for the isolation of collagen from fish scale by solvent extraction. The advancement from lab scale to pilot scale is a critical stage in the technological development. In this study, the optimal condition for the deproteinization which was validated at laboratory scale was employed in the pilot scale procedure. The deproteinization of fish scale was then demonstrated on a pilot scale (2Kg scales, 20l NaOH), resulting in protein content (0,2mg/ml) and hydroxyproline content (2,11mg/l). These results indicated that the pilot-scale showed similar performances to those of lab-scale one.

Keywords: deproteinization, pilot scale, scale, sardine pilchardus

Procedia PDF Downloads 420
323 Adaptive Certificate-Based Mutual Authentication Protocol for Mobile Grid Infrastructure

Authors: H. Parveen Begam, M. A. Maluk Mohamed

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Mobile Grid Computing is an environment that allows sharing and coordinated use of diverse resources in dynamic, heterogeneous and distributed environment using different types of electronic portable devices. In a grid environment the security issues are like authentication, authorization, message protection and delegation handled by GSI (Grid Security Infrastructure). Proving better security between mobile devices and grid infrastructure is a major issue, because of the open nature of wireless networks, heterogeneous and distributed environments. In a mobile grid environment, the individual computing devices may be resource-limited in isolation, as an aggregated sum, they have the potential to play a vital role within the mobile grid environment. Some adaptive methodology or solution is needed to solve the issues like authentication of a base station, security of information flowing between a mobile user and a base station, prevention of attacks within a base station, hand-over of authentication information, communication cost of establishing a session key between mobile user and base station, computing complexity of achieving authenticity and security. The sharing of resources of the devices can be achieved only through the trusted relationships between the mobile hosts (MHs). Before accessing the grid service, the mobile devices should be proven authentic. This paper proposes the dynamic certificate based mutual authentication protocol between two mobile hosts in a mobile grid environment. The certificate generation process is done by CA (Certificate Authority) for all the authenticated MHs. Security (because of validity period of the certificate) and dynamicity (transmission time) can be achieved through the secure service certificates. Authentication protocol is built on communication services to provide cryptographically secured mechanisms for verifying the identity of users and resources.

Keywords: mobile grid computing, certificate authority (CA), SSL/TLS protocol, secured service certificates

Procedia PDF Downloads 283
322 Simulation-Based Validation of Safe Human-Robot-Collaboration

Authors: Titanilla Komenda

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Human-machine-collaboration defines a direct interaction between humans and machines to fulfil specific tasks. Those so-called collaborative machines are used without fencing and interact with humans in predefined workspaces. Even though, human-machine-collaboration enables a flexible adaption to variable degrees of freedom, industrial applications are rarely found. The reasons for this are not technical progress but rather limitations in planning processes ensuring safety for operators. Until now, humans and machines were mainly considered separately in the planning process, focusing on ergonomics and system performance respectively. Within human-machine-collaboration, those aspects must not be seen in isolation from each other but rather need to be analysed in interaction. Furthermore, a simulation model is needed that can validate the system performance and ensure the safety for the operator at any given time. Following on from this, a holistic simulation model is presented, enabling a simulative representation of collaborative tasks – including both, humans and machines. The presented model does not only include a geometry and a motion model of interacting humans and machines but also a numerical behaviour model of humans as well as a Boole’s probabilistic sensor model. With this, error scenarios can be simulated by validating system behaviour in unplanned situations. As these models can be defined on the basis of Failure Mode and Effects Analysis as well as probabilities of errors, the implementation in a collaborative model is discussed and evaluated regarding limitations and simulation times. The functionality of the model is shown on industrial applications by comparing simulation results with video data. The analysis shows the impact of considering human factors in the planning process in contrast to only meeting system performance. In this sense, an optimisation function is presented that meets the trade-off between human and machine factors and aids in a successful and safe realisation of collaborative scenarios.

Keywords: human-machine-system, human-robot-collaboration, safety, simulation

Procedia PDF Downloads 338
321 Trends in the Incidence of Bloodstream Infections in Patients with Hematological Malignancies in the Period 1991–2012

Authors: V. N. Chebotkevich, E. E. Schetinkina, V. V. Burylev, E. I. Kaytandzhan, N. P. Stizhak

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Objective: Blood stream infections (BSI) are severe, life-threatening illness for immuno compromised patients with hematological malignancies. We report the trend in blood-stream infections in this group of patients in the period 1991-2013. Methods: A total of 4742 blood samples investigated. All blood cultures were incubated in a continuous monitoring system for 7 days before discarding negative. On signaled positive, organism was identified by conventional methods. The Real-time polymerase chain reaction (PCR) was used for the indication of human herpes virus 6 (HHV-6), Cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Results: Between 1991 and 2001 the incidence of Gram-positive bacteria (Staphylococcus epidermidis, Staphylococcus aureus) being the most common germs isolated (70,9%) were as Gram-negative rods (Escherichia coli, Klebsiella spp., Pseudomonas spp.) – 29,1%. In next decade 2002-2012 the number of Gram-negative bacteria was increased up to 40.2%. It is shown that the incidence of bacteremia was significantly more frequent at the background of detectable Cytomegalovirus and Epstein-Barr virus-specific DNA in blood. Over recent years, an increased frequency of micro mycetes was registered in blood of the patients with hematological malignancies (Candida spp. was predominant). Conclusion: Accurate and timely detection of BSI is important in determining appropriate treatment of infectious complications in patients with hematological malignancies. The isolation of Staphylococcus epidermidis from blood cultures remains a clinical dilemma for physicians and microbiologists. But in many cases this agent is of the clinical significance in immunocompromised patients with hematological malignancies. The role of CMV and EBV in development of bacteremia was demonstrated.

Keywords: infectious complications, blood stream infections, bacteremia, hemoblastosis

Procedia PDF Downloads 324
320 Harnessing the Generation of Ferromagnetic and Silver Nanostructures from Tropical Aquatic Microbial Nanofactories

Authors: Patricia Jayshree Jacob, Mas Jaffri Masarudinb, Mohd Zobir Hussein, Raha Abdul Rahim

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Iron based ferromagnetic nanoparticles (IONP) and silver nanostructures (AgNP) have found a wide range of application in antimicrobial therapy, cell targeting, and environmental applications. As such, the design of well-defined monodisperse IONPs and AgNPs have become an essential tool in nanotechnology. Fabrication of these nanostructures using conventional methods is not environmentally conducive and weigh heavily on energy and outlays. Selected microorganisms possess the innate ability to reduce metallic ions in colloidal aqueous solution to generate nanoparticles. Hence, harnessing this potential is a way forward in constructing microbial nano-factories, capable of churning out high yields of well-defined IONP’s and AgNP's with physicochemical characteristics on par with the best synthetically produced nanostructures. In this paper, we report the isolation and characterization of bacterial strains isolated from the tropical marine and freshwater ecosystems of Malaysia that demonstrated facile and rapid generation of ferromagnetic nanoparticles and silver nanostructures when precursors such as FeCl₃.6H₂O and AgNO₃ were added to the cell-free bacterial lysate in colloidal solution. Characterization of these nanoparticles was carried out using FESEM, UV Spectrophotometer, XRD, DLS and FTIR. This aerobic bioprocess was carried out at ambient temperature and humidity and has the potential to be developed for environmental friendly, cost effective large scale production of IONP’s. A preliminary bioprocess study on the harvesting time, incubation temperature and pH was also carried out to determine pertinent abiotic parameters contributing to the optimal production of these nanostructures.

Keywords: iron oxide nanoparticles, silver nanoparticles, biosynthesis, aquatic bacteria

Procedia PDF Downloads 257
319 Isolation and Expansion of Human Periosteum-Derived Mesenchymal Stem Cells in Defined Serum-Free Culture Medium

Authors: Ainur Mukhambetova, Miras Karzhauov, Vyacheslav Ogay

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Introduction: Mesenchymal stem cells (MSCs) have the capacity to be differentiated into several cell lineages and are a promising source for cell therapy and tissue engineering. However, currently most MSCs culturing protocols use media supplemented with fetal bovine serum (FBS), which limits their application in clinic due to the possibility of zoonotic infections, contamination and immunological reactions. Consequently, formulating effective serum free culture medium becomes one of the important problems in contemporary cell biotechnology. Objectives: The aim of this study was to define an optimal serum-free medium for culturing of periosteum derived MSCs. Materials and methods: The MSCs were extracted from human periosteum and transferred to the culture flasks pretreated with CELLstart™. Immunophenotypic characterization, proliferation and in vitro differentiation of cells grown on STEM PRO® MSC SFM were compared to the cells cultured in the standard FBS containing media. Chromosome analysis and flow cytometry were also performed. Results: We have shown that cells were grown on STEM PRO® MSC SFM retained all the morphological, immunophenotypic (CD73, CD90, CD105, vimentin and Stro-1) and cell differentiation characteristics specific to MSCs. Chromosome analysis indicated no anomalies in the chromosome structure. Flow cytometry showed a high expression of cell adhesion molecules CD44 (98,8%), CD90 (97,4%), CD105 (99,1%). In addition, we have shown that cell is grown on STEM PRO® MSC SFM have higher proliferation capacity compared to cell expanded on standard FBS containing the medium. Conclusion: We have shown that STEM PRO® MSC SFM is optimal for culturing periosteum derived human MSCs which subsequently can be safely used in cell therapy.

Keywords: cell technologies, periosteum-derived MSCs, regenerative medicine, serum-free medium

Procedia PDF Downloads 276