Search results for: zero knowledge protocol
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8326

Search results for: zero knowledge protocol

6436 Prediction of Embankment Fires at Railway Infrastructure Using Machine Learning, Geospatial Data and VIIRS Remote Sensing Imagery

Authors: Jan-Peter Mund, Christian Kind

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In view of the ongoing climate change and global warming, fires along railways in Germany are occurring more frequently, with sometimes massive consequences for railway operations and affected railroad infrastructure. In the absence of systematic studies within the infrastructure network of German Rail, little is known about the causes of such embankment fires. Since a further increase in these hazards is to be expected in the near future, there is a need for a sound knowledge of triggers and drivers for embankment fires as well as methodical knowledge of prediction tools. Two predictable future trends speak for the increasing relevance of the topic: through the intensification of the use of rail for passenger and freight transport (e.g..: doubling of annual passenger numbers by 2030, compared to 2019), there will be more rail traffic and also more maintenance and construction work on the railways. This research project approach uses satellite data to identify historical embankment fires along rail network infrastructure. The team links data from these fires with infrastructure and weather data and trains a machine-learning model with the aim of predicting fire hazards on sections of the track. Companies reflect on the results and use them on a pilot basis in precautionary measures.

Keywords: embankment fires, railway maintenance, machine learning, remote sensing, VIIRS data

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6435 Pluripotent Stem Cells as Therapeutic Tools for Limbal Stem Cell Deficiencies and Drug Testing

Authors: Aberdam Edith, Sangari Linda, Petit Isabelle, Aberdam Daniel

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Background and Rationale: Transparent avascularised cornea is essential for normal vision and depends on limbal stem cells (LSC) that reside between the cornea and the conjunctiva. Ocular burns or injuries may destroy the limbus, causing limbal stem cell deficiency (LSCD). The cornea becomes vascularised by invaded conjunctival cells, the stroma is scarring, resulting in corneal opacity and loss of vision. Grafted autologous limbus or cultivated autologous LCS can restore the vision, unless the two eyes are affected. Alternative cellular sources have been tested in the last decades, including oral mucosa or hair follicle epithelial cells. However, only partial success has been achieved by the use of these cells since they were not able to uniformly commit into corneal epithelial cells. Human pluripotent stem cells (iPSC) display both unlimited growth capacity and ability to differentiate into any cell type. Our goal was to design a standardized and reproducible protocol to produce transplantable autologous LSC from patients through cell reprogramming technology. Methodology: First, keratinocyte primary culture was established from a small number of plucked hair follicles of healthy donors. The resulting epithelial cells were reprogrammed into induced pluripotent stem cells (iPSCs) and further differentiate into corneal epithelial cells (CEC), according to a robust protocol that recapitulates the main step of corneal embryonic development. qRT-PCR analysis and immunofluorescent staining during the course of differentiation confirm the expression of stage specific markers of corneal embryonic lineage. First appear ectodermal progenitor-specific cytokeratins K8/K18, followed at day 7 by limbal-specific PAX6, TP63 and cytokeratins K5/K14. At day 15, K3/K12+-corneal cells are present. To amplify the iPSC-derived LSC (named COiPSC), intact small epithelial colonies were detached and cultivated in limbal cell-specific medium. In that culture conditions, the COiPSC can be frozen and thaw at any passage, while retaining their corneal characteristics for at least eight passages. To evaluate the potential of COiPSC as an alternative ocular toxicity model, COiPSC were treated at passage P0 to P4 with increasing amounts of SDS and Benzalkonium. Cell proliferation and apoptosis of treated cells was compared to LSC and the SV40-immortalized human corneal epithelial cell line (HCE) routinely used by cosmetological industrials. Of note, HCE are more resistant to toxicity than LSC. At P0, COiPSC were systematically more resistant to chemical toxicity than LSC and even to HCE. Remarkably, this behavior changed with passage since COiPSC at P2 became identical to LSC and thus closer to physiology than HCE. Comparative transcriptome analysis confirmed that COiPSC from P2 are similar to a mixture of LSC and CEC. Finally, by organotypic reconstitution assay, we demonstrated the ability of COiPSC to produce a 3D corneal epithelium on a stromal equivalent made of keratocytes. Conclusion: COiPSC could become valuable for two main applications: (1) an alternative robust tool to perform, in a reproducible and physiological manner, toxicity assays for cosmetic products and pharmacological tests of drugs. (2). COiPSC could become an alternative autologous source for cornea transplantation for LSCD.

Keywords: Limbal stem cell deficiency, iPSC, cornea, limbal stem cells

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6434 Tenure Security, Agricultural Diversity and Food Security

Authors: Amanuel Hadera Gebreyesus

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In the literature, the study of tenure and food security has largely involved separate lines of inquiry. In effect, the nexus among these has received little attention; and the underinvestment in research related to the relationship between tenure and food security deters generation of tenure-related knowledge and policy guidance for improving food and nutrition security. Drawing from this motivation, we study the relationship among tenure security, agricultural diversity and food security and dietary diversity. We employ IV approaches to examine the effect of tenure security and agricultural diversity on food security and dietary diversity. We find tenure security is inversely related with food insecurity as shown by its negative association with hunger scale, hunger index and hunger category. On the other hand, results suggest that tenure security improves minimum dietary diversity of women while we find no association with child dietary diversity. Moreover, agricultural diversity is positively related with minimum dietary diversity of women, which may point to higher accessibility and consumption of dietary food groups by women. Also, findings suggest that farmers use their human (knowledge and skills) and resource (land) endowments to improve food security and dietary diversity. An implication from this is the importance of not only improving access to land but also long-term tenure security to promote agricultural diversity, food security and dietary diversity.

Keywords: tenure security, food security, agricultural diversity, dietary diversity, women

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6433 Centering Critical Sociology for Social Justice and Inclusive Education

Authors: Al Karim Datoo

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Abstract— The presentation argues for an urgent case to center and integrate critical sociology in enriching potency of educational thought and practice to counteract inequalities and social injustices. COVID phenomenon has starkly exposed burgeoning of social-economic inequalities and widening marginalities which have been historically and politically constructed through deep-seated social and power imbalances and injustices in the world. What potent role could education possibly play to combat these issues? A point of departure for this paper highlights increasing reductionist and exclusionary ‘mind-set’ of education that has been developed through trends in education such as: the commodification of knowledge, standardisation, homogenization, and reification which are products of the positivist ideology of knowledge coopted to serve capitalist interests. To redress these issues of de-contextualization and de-humanization of education, it is emphasized that there is an urgent need to center the role of interpretive and critical epistemologies and pedagogies of social sciences. In this regard, notions of problem-posing versus problem-solving, generative themes, instrumental versus emancipatory reasoning will be discussed. The presentation will conclude by illustrating the pedagogic utility of these critically oriented notions to counteract the social reproduction of exclusionary and inequality in and through education.

Keywords: Critical pedagogy, social justice, inclusion , education

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6432 The Effect of Nursing Teamwork Training on Nursing Teamwork Effectiveness

Authors: Manar Ahmed Elbadawy

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Background: Empirical evidence suggested that improving nursing teamwork (NTW) may be the key to reducing medical error. The functioning nursing teams require open communication, mutual respect, and shared mental models to activate quality patient care. The complexity and the high demands for specialized nursing knowledge and skill also require nursing staff to consult with one another and work in teams regularly. The current study aimed to evaluate the effect of the nursing teamwork training program on nursing teamwork effectiveness. Design: A quasi-experimental (one group pretest-posttest) design was utilized. Three medical intensive care units at a teaching hospital affiliated to Cairo University Hospital, Egypt. Subjects: A convenient sample of 48 nursing staff worked at the selected units. The Nursing Teamwork Observational Checklist was used. Results: Total (NTW) mean scores exhibited quite elevation post-program implementation compared to preprogram and showed little decrease 3 months later ( = 2.52, SD = ± 0.27, mean % =51.98, = 2.72, SD = ± 0.20, mean %=72.45, = 2.67, SD = ± 0.11, mean %= 67.48 respectively). Conclusion: Implementation of (NTW) training program had a positive effect on increasing (NTW) effectiveness. Regular and frequent short-term teamwork training is important to be introduced as well as sustainable monitoring is required to ensure nursing attitudes, knowledge and skills’ change about teamwork effectiveness.

Keywords: effectiveness, nursing, teamwork, training

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6431 Exploring the Need to Study the Efficacy of VR Training Compared to Traditional Cybersecurity Training

Authors: Shaila Rana, Wasim Alhamdani

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Effective cybersecurity training is of the utmost importance, given the plethora of attacks that continue to increase in complexity and ubiquity. VR cybersecurity training remains a starkly understudied discipline. Studies that evaluated the effectiveness of VR cybersecurity training over traditional methods are required. An engaging and interactive platform can support knowledge retention of the training material. Consequently, an effective form of cybersecurity training is required to support a culture of cybersecurity awareness. Measurements of effectiveness varied throughout the studies, with surveys and observations being the two most utilized forms of evaluating effectiveness. Further research is needed to evaluate the effectiveness of VR cybersecurity training and traditional training. Additionally, research for evaluating if VR cybersecurity training is more effective than traditional methods is vital. This paper proposes a methodology to compare the two cybersecurity training methods and their effectiveness. The proposed framework includes developing both VR and traditional cybersecurity training methods and delivering them to at least 100 users. A quiz along with a survey will be administered and statistically analyzed to determine if there is a difference in knowledge retention and user satisfaction. The aim of this paper is to bring attention to the need to study VR cybersecurity training and its effectiveness compared to traditional training methods. This paper hopes to contribute to the cybersecurity training field by providing an effective way to train users for security awareness. If VR training is deemed more effective, this could create a new direction for cybersecurity training practices.

Keywords: virtual reality cybersecurity training, VR cybersecurity training, traditional cybersecurity training

Procedia PDF Downloads 206
6430 Animations for Teaching Food Chemistry: A Design Approach for Linking Chemistry Theory to Everyday Food

Authors: Paulomi (Polly) Burey, Zoe Lynch

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In STEM education, students often have difficulty linking static images and words from textbooks or online resources, to the underlying mechanisms of the topic of study. This can often dissuade some students from pursuing study in the physical and chemical sciences. A growing movement in current day students demonstrates that the YouTube generation feel they learn best from video or dynamic, interactive learning tools, and will seek these out as alternatives to their textbooks and the classroom learning environment. Chemistry, and in particular visualization of molecular structures in everyday materials, can prove difficult to comprehend without significant interaction with the teacher of the content and concepts, beyond the timeframe of a typical class. This can cause a learning hurdle for distance education students, and so it is necessary to provide strong electronic tools and resources to aid their learning. As one of the electronic resources, an animation design approach to link everyday materials to their underlying chemistry would be beneficial for student learning, with the focus here being on food. These animations were designed and storyboarded with a scaling approach and commence with a focus on the food material itself and its component parts. This is followed by animated transitions to its underlying microstructure and identifying features, and finally showing the molecules responsible for these microstructural features. The animation ends with a reverse transition back through the molecular structure, microstructure, all the way back to the original food material, and also animates some reactions that may occur during food processing to demonstrate the purpose of the underlying chemistry and how it affects the food we eat. Using this cyclical approach of linking students’ existing knowledge of food to help guide them to understanding more complex knowledge, and then reinforcing their learning by linking back to their prior knowledge again, enhances student understanding. Food is also an ideal material system for students to interact with, in a hands-on manner to further reinforce their learning. These animations were launched this year in a 2nd year University Food Chemistry course with improved learning outcomes for the cohort.

Keywords: chemistry, food science, future pedagogy, STEM Education

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6429 Regional Barriers and Opportunities for Developing Innovation Networks in the New Media Industry: A Comparison between Beijing and Bangalore Regional Innovation Systems

Authors: Cristina Chaminade, Mandar Kulkarni, Balaji Parthasarathy, Monica Plechero

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The characteristics of a regional innovation system (RIS) and the specificity of the knowledge base of an industry may contribute to create peculiar paths for innovation and development of firms’ geographic extended innovation networks. However, the relative empirical evidence in emerging economies remains underexplored. The paper aims to fill the research gap by means of some recent qualitative research conducted in 2016 in Beijing (China) and Bangalore (India). It analyzes cases studies of firms in the new media industry, a sector that merges different IT competences with competences from other knowledge domains and that is emerging in those RIS. The results show that while in Beijing the new media sector results to be more in line with the existing institutional setting and governmental goals aimed at targeting specific social aspects and social problems of the population, in Bangalore it remains a more spontaneous firms-led process. In Beijing what matters for the development of innovation networks is the governmental setting and the national and regional strategies to promote science and technology in this sector, internet and mass innovation. The peculiarities of recent governmental policies aligned to the domestic goals may provide good possibilities for start-ups to develop innovation networks. However, due to the specificities of those policies targeting the Chinese market, networking outside the domestic market are not so promoted. Moreover, while some institutional peculiarities, such as a culture of collaboration in the region, may be favorable for local networking, regulations related to Internet censorship may limit the use of global networks particularly when based on virtual spaces. Mainly firms with already some foreign experiences and contact take advantage of global networks. In Bangalore, the role of government in pushing networking for the new media industry at the present stage is quite absent at all geographical levels. Indeed there is no particular strategic planning or prioritizing in the region toward the new media industry, albeit one industrial organization has emerged to represent the animation industry interests. This results in a lack of initiatives for sustaining the integration of complementary knowledge into the local portfolio of IT specialization. Firms actually involved in the new media industry face institutional constrains related to a poor level of local trust and cooperation, something that does not allow for full exploitation of local linkages. Moreover, knowledge-provider organizations in Bangalore remain still a solid base for the IT domain, but not for other domains. Initiatives to link to international networks seem therefore more the result of individual entrepreneurial actions aimed at acquiring complementary knowledge and competencies from different domains and exploiting potentiality in different markets. From those cases, it emerges that role of government, soft institutions and organizations in the two RIS differ substantially in the creation of barriers and opportunities for the development of innovation networks and their specific aim.

Keywords: regional innovation system, emerging economies, innovation network, institutions, organizations, Bangalore, Beijing

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6428 Artificial Intelligence Based Method in Identifying Tumour Infiltrating Lymphocytes of Triple Negative Breast Cancer

Authors: Nurkhairul Bariyah Baharun, Afzan Adam, Reena Rahayu Md Zin

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Tumor microenvironment (TME) in breast cancer is mainly composed of cancer cells, immune cells, and stromal cells. The interaction between cancer cells and their microenvironment plays an important role in tumor development, progression, and treatment response. The TME in breast cancer includes tumor-infiltrating lymphocytes (TILs) that are implicated in killing tumor cells. TILs can be found in tumor stroma (sTILs) and within the tumor (iTILs). TILs in triple negative breast cancer (TNBC) have been demonstrated to have prognostic and potentially predictive value. The international Immune-Oncology Biomarker Working Group (TIL-WG) had developed a guideline focus on the assessment of sTILs using hematoxylin and eosin (H&E)-stained slides. According to the guideline, the pathologists use “eye balling” method on the H&E stained- slide for sTILs assessment. This method has low precision, poor interobserver reproducibility, and is time-consuming for a comprehensive evaluation, besides only counted sTILs in their assessment. The TIL-WG has therefore recommended that any algorithm for computational assessment of TILs utilizing the guidelines provided to overcome the limitations of manual assessment, thus providing highly accurate and reliable TILs detection and classification for reproducible and quantitative measurement. This study is carried out to develop a TNBC digital whole slide image (WSI) dataset from H&E-stained slides and IHC (CD4+ and CD8+) stained slides. TNBC cases were retrieved from the database of the Department of Pathology, Hospital Canselor Tuanku Muhriz (HCTM). TNBC cases diagnosed between the year 2010 and 2021 with no history of other cancer and available block tissue were included in the study (n=58). Tissue blocks were sectioned approximately 4 µm for H&E and IHC stain. The H&E staining was performed according to a well-established protocol. Indirect IHC stain was also performed on the tissue sections using protocol from Diagnostic BioSystems PolyVue™ Plus Kit, USA. The slides were stained with rabbit monoclonal, CD8 antibody (SP16) and Rabbit monoclonal, CD4 antibody (EP204). The selected and quality-checked slides were then scanned using a high-resolution whole slide scanner (Pannoramic DESK II DW- slide scanner) to digitalize the tissue image with a pixel resolution of 20x magnification. A manual TILs (sTILs and iTILs) assessment was then carried out by the appointed pathologist (2 pathologists) for manual TILs scoring from the digital WSIs following the guideline developed by TIL-WG 2014, and the result displayed as the percentage of sTILs and iTILs per mm² stromal and tumour area on the tissue. Following this, we aimed to develop an automated digital image scoring framework that incorporates key elements of manual guidelines (including both sTILs and iTILs) using manually annotated data for robust and objective quantification of TILs in TNBC. From the study, we have developed a digital dataset of TNBC H&E and IHC (CD4+ and CD8+) stained slides. We hope that an automated based scoring method can provide quantitative and interpretable TILs scoring, which correlates with the manual pathologist-derived sTILs and iTILs scoring and thus has potential prognostic implications.

Keywords: automated quantification, digital pathology, triple negative breast cancer, tumour infiltrating lymphocytes

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6427 Biodiversity Conservation Practices Among Indigenous Peoples in Caraga Region, Mindanao, Philippines

Authors: Milagros S. Salibad, Levita B. Grana

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The presence and role of Indigenous Peoples residing in key biodiversity, protected, and watershed areas within the ancestral domain in the Caraga Region hold immense significance. This study aimed to determine the level of biodiversity conservation practices among the Mamanwas, Manobos, and Higaonons, and identify facilitating or hindering factors. Employing a mixed-method research design, 421 respondents participated through a researcher-made questionnaire. Focus group discussions, key informant interviews, researcher field notes, community immersions, and secondary sources were done. The three groups have demonstrated a high level of biodiversity conservation practices manifesting their commitment to conserving their natural resources and ecosystems. Evidently, selecting and cutting only mature trees for shelter and tribal usage, and preservation of large trees that harbor ancestors’ spirits and worship through rituals (Mambabaja). Each group exhibited unique environmental practices shaped by their distinct cultures, traditions, customary knowledge, and access to information. The Mamanwa practiced traditional hunting and gathering by using traps while Manobo practiced shifting cultivation to maintain soil fertility and biodiversity, and Higaonon managed forest resources through traditional forest management (establishment of sacred forests and conservation areas). Various facilitating and hindering factors influenced their conservation efforts. Their traditional knowledge and practices, partnership and collaboration, legal recognition and support, access to information, and biodiversity monitoring system facilitate practices. Insufficient government assistance, political and social issues, scarce financial support, inadequate policy enforcement, lack of livelihood opportunities, and land use conflicts hinder them. Monitoring the sustainability of IPs' local biodiversity conservation practices is essential as they contribute to conservation endeavors.

Keywords: biodiversity, conservation, indigenous peoples, traditional knowledge

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6426 A Basic Understanding of Viral Disease and Education Level Influences Disease Risk Perception, Disease Severity Perception, and Mask Wearing Behavior During the COVID-19 Pandemic

Authors: Ilse Kreme

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To the best of this author’s knowledge, no studies have been identified on the connection between a refusal to engage in health-protective behaviors and a basic understanding of viral biology among community college students, faculty, and staff during the COVID-19 pandemic. Lack of scientific knowledge could prevent understanding of why these behaviors are important to prevent the community spread of COVID-19, even when they are not shown to offer much individual protection. In this study, a possible correlation was examined between a basic knowledge level of viral disease that comes from having taken a college biology course and disease perceptions of COVID-19. In particular, disease risk perception, disease severity percept and mask-wearing behaviors were examined as they correlated with having taken an undergraduate biology course. The effect of covariates of age, gender, and education level were investigated along with the main dependent variables. A representative sample of the population included students, faculty, and staff at Paradise Valley Community College (PVCC) in Phoenix, Arizona. Participants were recruited by an email sent to all students, faculty, and staff at PVCC using an all-college email distribution. Disease risk and severity perception were assessed with the Brief Illness Perception Questionnaire 5 (BIP-Q5), which was modified to include questions measuring participant age, education level, and whether they took or ever took a college biology course. Two additional questions measured compliance of willingness to wear a face mask. The results showed an effect of gender on mask-wearing behavior and a correlation between having taken a biology course and disease severity perception. No differences were seen in mask-wearing behavior and disease risk perception as a result of having taken a biology course. These findings suggest that taking an undergraduate biology course leads to a greater awareness of COVID-19 disease severity through an understanding of the basic biological principles of viral disease transmission. The results can be used to modify existing health education strategies. Further research is needed on how to best reach target audiences in all education brackets.

Keywords: COVID-19, education, gender, mask wearing, disease risk perception, disease severity perception

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6425 The Effectiveness of Cathodic Protection on Microbiologically Influenced Corrosion Control

Authors: S. Taghavi Kalajahi, A. Koerdt, T. Lund Skovhus

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Cathodic protection (CP) is an electrochemical method to control and manage corrosion in different industries and environments. CP which is widely used, especially in buried and sub-merged environments, which both environments are susceptible to microbiologically influenced corrosion (MIC). Most of the standards recommend performing CP using -800 mV, however, if MIC threats are high or sulfate reducing bacteria (SRB) is present, the recommendation is to use more negative potentials for adequate protection of the metal. Due to the lack of knowledge and research on the effectiveness of CP on MIC, to the author’s best knowledge, there is no information about what MIC threat is and how much more negative potentials should be used enabling adequate protection and not overprotection (due to hydrogen embrittlement risk). Recently, the development and cheaper price of molecular microbial methods (MMMs) open the door for more effective investigations on the corrosion in the presence of microorganisms, along with other electrochemical methods and surface analysis. In this work, using MMMs, the gene expression of SRB biofilm under different potentials of CP will be investigated. The specific genes, such as pH buffering, metal oxidizing, etc., will be compared at different potentials, enabling to determine the precise potential that protect the metal effectively from SRB. This work is the initial step to be able to standardize the recommended potential under MIC condition, resulting better protection for the infrastructures.

Keywords: cathodic protection, microbiologically influenced corrosion, molecular microbial methods, sulfate reducing bacteria

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6424 Influence of the Local External Pressure on Measured Parameters of Cutaneous Microcirculation

Authors: Irina Mizeva, Elena Potapova, Viktor Dremin, Mikhail Mezentsev, Valeri Shupletsov

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The local tissue perfusion is regulated by the microvascular tone which is under the control of a number of physiological mechanisms. Laser Doppler flowmetry (LDF) together with wavelet analyses is the most commonly used technique to study the regulatory mechanisms of cutaneous microcirculation. External factors such as temperature, local pressure of the probe on the skin, etc. influence on the blood flow characteristics and are used as physiological tests to evaluate microvascular regulatory mechanisms. Local probe pressure influences on the microcirculation parameters measured by optical methods: diffuse reflectance spectroscopy, fluorescence spectroscopy, and LDF. Therefore, further study of probe pressure effects can be useful to improve the reliability of optical measurement. During pressure tests variation of the mean perfusion measured by means of LDF usually is estimated. An additional information concerning the physiological mechanisms of the vascular tone regulation system in response to local pressure can be obtained using spectral analyses of LDF samples. The aim of the present work was to develop protocol and algorithm of data processing appropriate for study physiological response to the local pressure test. Involving 6 subjects (20±2 years) and providing 5 measurements for every subject we estimated intersubject and-inter group variability of response of both averaged and oscillating parts of the LDF sample on external surface pressure. The final purpose of the work was to find special features which further can be used in wider clinic studies. The cutaneous perfusion measurements were carried out by LAKK-02 (SPE LAZMA Ltd., Russia), the skin loading was provided by the originally designed device which allows one to distribute the pressure around the LDF probe. The probe was installed on the dorsal part of the distal finger of the index figure. We collected measurements continuously for one hour and varied loading from 0 to 180mmHg stepwise with a step duration of 10 minutes. Further, we post-processed the samples using the wavelet transform and traced the energy of oscillations in five frequency bands over time. Weak loading leads to pressure-induced vasodilation, so one should take into account that the perfusion measured under pressure conditions will be overestimated. On the other hand, we revealed a decrease in endothelial associated fluctuations. Further loading (88 mmHg) induces amplification of pulsations in all frequency bands. We assume that such loading leads to a higher number of closed capillaries, higher input of arterioles in the LDF signal and as a consequence more vivid oscillations which mainly are formed in arterioles. External pressure higher than 144 mmHg leads to the decrease of oscillating components, after removing the loading very rapid restore of the tissue perfusion takes place. In this work, we have demonstrated that local skin loading influence on the microcirculation parameters measured by optic technique; this should be taken into account while developing portable electronic devices. The proposed protocol of local loading allows one to evaluate PIV as far as to trace dynamic of blood flow oscillations. This study was supported by the Russian Science Foundation under project N 18-15-00201.

Keywords: blood microcirculation, laser Doppler flowmetry, pressure-induced vasodilation, wavelet analyses blood

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6423 Nyaya, Buddhist School Controversy regarding the Laksana of Pratyaksa: Causal versus Conceptual Analysis

Authors: Maitreyee Datta

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Buddhist lakṣaņa of pratyakṣa pramā is not the result of the causal analysis of the genesis of it. Naiyāyikas, on the other hand, has provided the lakṣaņa of pratyakṣa in terms of the causal analysis of it. Thus, though in these two philosophical systems philosophers have discussed in detail the nature of pratyakṣa pramā (perception), yet their treatments and understanding of it vary according to their respective understanding of pramā and prmāņa and their relationship. In Nyāya school, the definition (lakṣņa) of perception (pratyakṣa) has been given in terms of the process by virtue of which it has been generated. Thus, Naiyāyikas were found to provide a causal account of perception (pratyakṣa) by virtue of their lakṣaņa of it. But in Buddhist epistemology perception has been defined by virtue of the nature of perceptual knowledge (pratyakṣa pramā) which is devoid of any vikalpa or cognition. These two schools differed due to their different metaphysical presuppositions which determine their epistemological pursuits. The Naiyāyikas admitted pramā and pramāņa as separate events and they have taken pramāņa to be the cause of pramā. These presuppositions enabled them to provide a lakṣaņa of pratyakṣa pramā in terms of the causes by which it is generated. Why did the Buddhist epistemologists define perception by the unique nature of perceptual knowledge instead of the process by which it is generated? This question will be addressed and dealt with in the present paper. In doing so, the unique purpose of Buddhist philosophy will be identified which will enable us to find out an answer to the above question. This enterprise will also reveal the close relationship among some basic Buddhist presuppositions like pratityasamutpādavāda and kṣaņikavāda with Buddhist epistemological positions. In other words, their distinctive notion of pramā (knowledge) indicates their unique epistemological position which is found to comply with their basic philosophical presuppositions. The first section of the paper will present the Buddhist epistemologists’ lakṣaņa of pratyakṣa. The analysis of the lakṣaņa will be given in clear terms to reveal the nature of pratyakṣa as an instance of pramā. In the second section, an effort will be made to identify the uniqueness of such a definition. Here an articulation will be made in which the relationship among basic Buddhist presuppositions and their unique epistemological positions are determined. In the third section of the paper, an effort will be made to compare Nyāya epistemologist’s position regarding pratyakṣa with that of the Buddhist epistemologist.

Keywords: laksana, prama, pramana, pratyksa

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6422 Biological Hazards and Laboratory inflicted Infections in Sub-Saharan Africa

Authors: Godfrey Muiya Mukala

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This research looks at an array of fields in Sub-Saharan Africa comprising agriculture, food enterprises, medicine, organisms genetically modified, microbiology, and nanotechnology that can be gained from biotechnological research and development. Findings into dangerous organisms, mainly bacterial germs, rickettsia, fungi, parasites, or organisms that are genetically engineered, have immensely posed questions attributed to the biological danger they bring forth to human beings and the environment because of their uncertainties. In addition, the recurrence of previously managed diseases or the inception of new diseases are connected to biosafety challenges, especially in rural set-ups in low and middle-income countries. Notably, biotechnology laboratories are required to adopt biosafety measures to protect their workforce, community, environment, and ecosystem from unforeseen materials and organisms. Sensitization and inclusion of educational frameworks for laboratory workers are essential to acquiring a solid knowledge of harmful biological agents. This is in addition to human pathogenicity, susceptibility, and epidemiology to the biological data used in research and development. This article reviews and analyzes research intending to identify the proper implementation of universally accepted practices in laboratory safety and biological hazards. This research identifies ideal microbiological methods, adequate containment equipment, sufficient resources, safety barriers, specific training, and education of the laboratory workforce to decrease and contain biological hazards. Subsequently, knowledge of standardized microbiological techniques and processes, in addition to the employment of containment facilities, protective barriers, and equipment, is far-reaching in preventing occupational infections. Similarly, reduction of risks and prevention may be attained by training, education, and research on biohazards, pathogenicity, and epidemiology of the relevant microorganisms. In this technique, medical professionals in rural setups may adopt the knowledge acquired from the past to project possible concerns in the future.

Keywords: sub-saharan africa, biotechnology, laboratory, infections, health

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6421 Effect of Unilateral Unoperated Ovarian Endometrioma on Responsiveness to Hyperstimulation

Authors: Abdelmaguid Ramzy, Mohamed Bahaa

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Introduction: The effects of ovarian endometrioma on fertility outcomes with IVF have been always related to poor outcomes. Objective: To evaluate the effect of unilateral unoperated ovarian endometrioma < 2cm on the number of developing follicles and compare them with the contralateral ovary as a control. Design: Retrospective case control study. Setting: KasrEl-Aini IVF center. Patient(s): We studied 32 women with unilateral endometrioma who underwent their first IVF cycle. Methods: 32 Patients aged between 20-35 years selected for IVF who were diagnosed with one unilateral endometrioma (diameter <2 cm) and who did not undergo previous ovarian surgery were retrospectively identified. The number of follicles > 17 mm during folliculometry on the day of HCG trigger in the ovary that contained endometrioma were compared with those from the contralateral ovary. They were all hyperstimulated using long protocol with (225-300 IU) gonadotrophins. Primary outcome: The number of follicles > 17 mm during folliculometry on the day of HCG trigger. Result(s): The mean ± SD age, Day 3 serum FSH and LH were 27± 3.7 years, 5.8 ± 1.6 IU/ml and 4.5 ± 1.7 IU/ml respectively. There was no significant difference in the number of follicles > 17 mm on the day of HCG trigger in the ovary that contained endometrioma (4.4 ±2.5) and in the opposite ovary (4.5 ± 2.8) (P= 0.48). Conclusion: The presence of ovarian endometrioma in a controlled ovarian hyperstimulation cycle for IVF treatment is not associated with a reduced number of follicles > 17 mm during folliculometry on the day of HCG trigger.

Keywords: endometrioma, folliculometry, hyperstimulation, fertility

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6420 Women Executives' Career Success in the Office of the Basic Education

Authors: Nipon Sasithornsaowapa

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This research aims to study the impact of personality and family status on women executives’ career success of the primary education department of Thailand. The independent variable includes three factors, namely family status, personality, and knowledge-skill-experience, while the dependent variable is the career success. The population of this study includes 2,179 female management officials in the department of primary education. A total of 400 female managers is interviewed and utilized as a sample group. A questionnaire is developed and used as a research tool for data collection. Content analysis is performed to get the quantitative data. Descriptive statistics in this research is conducted by SPSS program. The findings revealed that personality and family status of samples have an influence on the overall career success of women executives in terms of their objective career success. However, in terms of specific factors of personality or family status, it is found that there is no relevance of each factor on the women executives’ career success. It can be concluded that the factor affecting the women executives’ career success is subjective career success including the happiness and enjoyment with the job not factor concerning materials. Their success is the result of each individual working experience. However, their personal characteristics do not affect their success.

Keywords: career success, women executives, primary education, knowledge-skill-experience

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6419 Sustainable Design through up-Cycling Crafts in the Mainstream Fashion Industry of India

Authors: Avani Chhajlani

Abstract:

Fashion is considered to be the most destructive industry, second only to the oil rigging industry, which has a greater impact on the environment. While fashion today banks upon fast fashion to generate a higher turnover of designs and patterns in apparel and related accessories, crafts push us towards a more slow and thoughtful approach with culturally identifiably unique work and slow community-centered production. Despite this strong link between indigenous crafts and sustainability, it has not been extensively researched and explored upon. In the forthcoming years, the fashion industry will have to reinvent itself to move towards a more holistic and sustainable circular model to balance the harm already caused. And closed loops of the circular economy will help the integration of indigenous craft knowledge, which is regenerative. Though sustainability and crafts of a region go hand-in-hand, the craft still have to find its standing in the mainstream fashion world; craft practices have a strong local congruence and knowledge that has been passed down generation-to-generation through oration or written materials. This paper aims to explore ways a circular economy can be created by amalgamating fashion and craft while creating a sustainable business model and how this is slowly being created today through brands like – RaasLeela, Pero, and KaSha, to name a few.

Keywords: circular economy, fashion, India, indigenous crafts, slow fashion, sustainability, up-cycling

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6418 Effects of Elastic, Plyometric and Strength Training on Selected Anaerobic Factors in Sanandaj Elite Volleyball Players

Authors: Majed Zobairy, Fardin Kalvandi, Kamal Azizbaigi

Abstract:

This research was carried out for evaluation of elastic, plyometric and resistance training on selected anaerobic factors in men volleyball players. For these reason 30 elite volleyball players of Sanandaj city randomly divided into 3 groups as follow: elastic training, plyometric training and resistance training. Pre-exercise tests which include vertical jumping, 50 yard speed running and scat test were done and data were recorded. Specific exercise protocol regimen was done for each group and then post-exercise tests again were done. Data analysis showed that there were significant increases in exercise test in each group. One way ANOVA analysis showed that increases in speed records in elastic group were significantly higher than the other groups (p<0/05),based on research data it seems that elastic training can be a useful method and new approach in improving functional test and training regimen.

Keywords: elastic training, plyometric training, strength training, anaerobic power

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6417 Testing Two Actors Contextual Interaction Theory in a Multi Actors Context: Case of COVID-19 Disease Prevention and Control Policy

Authors: Muhammad Fayyaz Nazir, Ellen Wayenberg, Shahzadaah Faahed Qureshi

Abstract:

Introduction: The study is based on the Contextual Interaction Theory (CIT) constructs to explore the role of policy actors in implementing the COVID-19 Disease Prevention and Control (DP&C) Policy. The study analyzes the role of healthcare workers' contextual factors, such as cognition, motives, and resources, and their interactions in implementing Social Distancing (SD). In this way, we test a two actors policy implementation theory, i.e., the CIT in a three-actor context. Methods: Data was collected through document analysis and semi-structured interviews. For a qualitative study design, interviews were conducted with questions on cognition, motives, and resources from the healthcare workers involved in implementing SD in the local context in Multan – Pakistan. The possible interactions resulting from contextual factors of the policy actors – healthcare workers were identified through framework analysis protocol guided by CIT and supported by trustworthiness criterion and data saturation. Results: This inquiry resulted in theory application, addition, and enrichment. The theoretical application in the three actor's contexts illustrates the different levels of motives, cognition, and resources of healthcare workers – senior administrators, managers, and healthcare professionals. The senior administrators working in National Command and Operations Center (NCOC), Provincial Technical Committees (PTCs), and Districts Covid Teams (DCTs) were playing their role with high motivation. They were fully informed about the policy and moderately resourceful. The policy implementors: healthcare managers working on implementing the SD within their respective hospitals were playing their role with high motivation and were fully informed about the policy. However, they lacked the required resources to implement SD. The target medical and allied healthcare professionals were moderately motivated but lack of resources and information. The interaction resulted in cooperation and the need for learning to manage the future healthcare crisis. However, the lack of resources created opposition to the implementation of SD. Objectives of the Study: The study aimed to apply a two actors theory in a multi actors context. We take this as an opportunity to qualitatively test the theory in a novel situation of the Covid-19 pandemic and make way for its quantitative application by designing a survey instrument so that implementation researchers can apply CIT through multivariate analyses or higher-order statistical modeling. Conclusion: Applying two actors' implementation theory in exploring a complex case of healthcare intervention in three actors context is a unique work that has never been done before, up to the best of our knowledge. So, the work will contribute to the policy implementation studies by applying, extending, and enriching an implementation theory in a novel case of the Covi-19 pandemic, ultimately fulfilling the gap in implementation literature. Policy institutions and other low or middle-income countries can learn from this research and improve SD implementation by working on the variables with weak significance levels.

Keywords: COVID-19, disease prevention and control policy, implementation, policy actors, social distancing

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6416 Energy-Efficient Contact Selection Method for CARD in Wireless Ad-Hoc Networks

Authors: Mehdi Assefi, Keihan Hataminezhad

Abstract:

One of the efficient architectures for exploring the resources in wireless ad-hoc networks is contact-based architecture. In this architecture, each node assigns a unique zone for itself and each node keeps all information from inside the zone, as well as some from outside the zone, which is called contact. Reducing the overlap between different zones of a node and its contacts increases its performance, therefore Edge Method (EM) is designed for this purpose. Contacts selected by EM do not have any overlap with their sources, but for choosing the contact a vast amount of information must be transmitted. In this article, we will offer a new protocol for contact selection, which is called PEM. The objective would be reducing the volume of transmitted information, using Non-Uniform Dissemination Probabilistic Protocols. Consumed energy for contact selection is a function of the size of transmitted information between nodes. Therefore, by reducing the content of contact selection message using the PEM will decrease the consumed energy. For evaluation of the PEM we applied the simulation method. Results indicated that PEM consumes less energy compared to EM, and by increasing the number of nodes (level of nodes), performance of PEM will improve in comparison with EM.

Keywords: wireless ad-hoc networks, contact selection, method for CARD, energy-efficient

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6415 Alternating Electric fields-Induced Senescence in Glioblastoma

Authors: Eun Ho Kim

Abstract:

Innovations have conjured up a mode of treating GBM cancer cells in the newly diagnosed patients in a period of 4.9 months at an improved median OS, which brings along only a few minor side effects in the phase III of the clinical trial. This mode has been termed the Alternating Electric Fields (AEF). The study at hand is aimed at determining whether the AEF treatment is beneficial in sensitizing the GBM cancer cells through the process of increasing the AEF –induced senescence. The methodology to obtain the findings for this research ranged across various components, such as obtaining and testing SA-β-gal staining, flow cytometry, Western blotting, morphology, and Positron Emission Tomography (PET) / Computed Tomography (CT), immunohistochemical staining and microarray. The number of cells that displayed a senescence-specific morphology and positive SA-ß-Gal activity gradually increased up to 5 days. These results suggest that p16, p21 and p27 are essential regulators of AEF -induced senescence via NF-κB activation. The results showed that the AEF treatment is functional in enhancing the AEF –induced senescence in the GBM cells via an apoptosis- independent mechanism. This research concludes that this mode of treatment is a trustworthy protocol that can be effectively employed to overcome the limitations of the conventional mode of treatment on GBM.

Keywords: alternating electric fields, senescence, glioblastoma, cell death

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6414 Traditional Management Systems and the Conservation of Cultural and Natural Heritage: Multiple Case Studies in Zimbabwe

Authors: Nyasha Agnes Gurira, Petronella Katekwe

Abstract:

Traditional management systems (TMS) are a vital source of knowledge for conserving cultural and natural heritage. TMS’s are renowned for their ability to preserve both tangible and intangible manifestations of heritage. They are a construct of the intricate relationship that exists between heritage and host communities, where communities are recognized as owners of heritage and so, set up management mechanisms to ensure its adequate conservation. Multiple heritage condition surveys were conducted to assess the effectiveness of using TMS in the conservation of both natural and cultural heritage. Surveys were done at Nharira Hills, Mahwemasimike, Dzimbahwe, Manjowe Rock art sites and Norumedzo forest which are heritage places in Zimbabwe. It assessed the state of conservation of the five case studies and assessed the role that host communities play in the management of these heritage places. It was revealed that TMS’s are effective in the conservation of natural heritage, however in relation to heritage forms with cultural manifestations, there are major disparities. These range from differences in appreciation and perception of value within communities leading to vandalism, over emphasis in the conservation of the intangible element as opposed to the tangible. This leaves the tangible element at risk. Despite these issues, TMS are a reliable knowledge base which enables more holistic conservation approaches for cultural and natural heritage.

Keywords: communities, cultural intangible, tangible heritage, traditional management systems, natural

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6413 Lifelong Learning and Digital Literacies in Language Learning

Authors: Selma Karabinar

Abstract:

Lifelong learning can be described as a system where learning takes place for a person over the course of a lifespan and comprises formal, non-formal and informal learning to achieve the maximum possible improvement in personal, social, and vocational life. 21st century is marked with the digital technologies and people need to learn and adapt to new literacies as part of their lifelong learning. Our current knowledge gap brings to mind several questions: Do people with digital mindsets have different assumptions about affordances of digital technologies? How do digital mindsets lead language learners use digital technologies within and beyond classrooms? Does digital literacies have different significance for the learners? The presentation is based on a study attempted to answer these questions and show the relationship between lifelong learning and digital literacies. The study was conducted with learners of English language at a state university in Istanbul. The quantitative data in terms of participants' lifelong learning perception was collected through a lifelong learning scale from 150 students. Then 5 students with high and 5 with low lifelong learning perception were interviewed. They were questioned about their personal sense of agency in lifelong learning and how they use digital technologies in their language learning. Therefore, the qualitative data was analyzed in terms of their knowledge about digital literacies and actual use of it in their personal and educational life. The results of the study suggest why teaching new literacies are important for lifelong learning and also suggests implications for language teachers' education and language pedagogy.

Keywords: digital mindsets, language learning, lifelong learning, new literacies

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6412 Children Protection in the Digital Space

Authors: Beverly Komen

Abstract:

Online crimes have been on the rise in the recent days, especially with the hit of the covid-19 pandemic. The coronavirus pandemic has led to an unprecedented rise in screen time, this means more families are relying on technology and digital solutions to keep children learning, spending more time on the virtual platforms can leave children vulnerable to online abuse and exploitation. With ease access of affordable phones, internet, and increased online activities, all children are at risk of being abused online hence making the digital space unsafe for children. With these increased use of technology and its accessibility, children are at risk of facing challenges such as access to inappropriate content, online grooming, identity theft, cyber bullying, among other risks. The big question is; as we enjoy the benefits brought in by technology, how do we ensure that our children are save in this digital space? With the analysis of the current trends, there is a gap in knowledge on people’s understanding on child online protection and safety measures when using the digital space. A survey conducted among 50 parents in Nairobi in Kenya indicated that there is a gap in knowledge on online protection of children and over 50 % of the participants shared that for sure they have no idea on how to protect children online. This paper seeks to address the concept of child protection in the digital space and come up with viable solutions in protecting children from online vices.

Keywords: child protection, digital space, online risks, online grooming, cyber bulying, online child sexual exploitation, and abuse

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6411 Conceptual Model for Massive Open Online Blended Courses Based on Disciplines’ Concepts Capitalization and Obstacles’ Detection

Authors: N. Hammid, F. Bouarab-Dahmani, T. Berkane

Abstract:

Since its appearance, the MOOC (massive open online course) is gaining more and more intention of the educational communities over the world. Apart from the current MOOCs design and purposes, the creators of MOOC focused on the importance of the connection and knowledge exchange between individuals in learning. In this paper, we present a conceptual model for massive open online blended courses where teachers over the world can collaborate and exchange their experience to get a common efficient content designed as a MOOC opened to their students to live a better learning experience. This model is based on disciplines’ concepts capitalization and the detection of the obstacles met by their students when faced with problem situations (exercises, projects, case studies, etc.). This detection is possible by analyzing the frequently of semantic errors committed by the students. The participation of teachers in the design of the course and the attendance by their students can guarantee an efficient and extensive participation (an important number of participants) in the course, the learners’ motivation and the evaluation issues, in the way that the teachers designing the course assess their students. Thus, the teachers review, together with their knowledge, offer a better assessment and efficient connections to their students.

Keywords: massive open online course, MOOC, online learning, e-learning

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6410 Infant and Young Child Dietary Diversification Using Locally Available Foods after Nutrition Education in Rural Malawi

Authors: G. C. Phiri, E. A. Heil, A. A. Kalimbira, E. Muehlhoff, C. Masangano, B. M. Mtimuni, J. Herrmann, M. B. Krawinkel, I. Jordan

Abstract:

Background and objectives: High prevalence of undernutrition in Malawi is caused by poor complementary foods. Lack of knowledge of age appropriate food within the household might affect utilization of available resources. FAO-Malawi implemented nutrition education (NE) sessions in 200 villages in Kasungu and Mzimba districts from December 2012 to April 2013 targeting 15 caregivers per village of children aged 6-18 months, grandmothers, spouses and community leaders. Two trained volunteers per village facilitated 10 NE sessions on breastfeeding, food safety and hygiene and complementary feeding using locally available resources. This study assessed the reported dietary diversification practices of infant and young child after nutrition education and the factors that influenced adoption of the practice. Methodology: Questionnaire-based interviews with caregivers were conducted in 16 randomly selected villages (n=108) before training-(t1) and seven months after training-(t2). Knowledge score (KS) was calculated on the indicators breastfeeding, hygiene and complementary feeding. Count regression was performed using SPSS 22. Eight focus group discussions (FGDs) were separately conducted among caregivers and grandmothers in 4 villages. Content analysis was used to analyze FGDs data. Results: Following NE, caregivers' KS significantly increased (p<0.001) between t1 and t2 for breastfeeding (7.7 vs. 9.8, max=18), hygiene (3.8 vs. 5.9, max=7) and complementary feeding (10.2 vs. 16.2, max=26). Caregivers indicated that they stopped preparation of plain-refined maize meal porridge after they gained knowledge on dietary diversification of complementary foods. They learnt mushing and pounding of ingredients for enriched porridge. Whole-maize meal or potatoes were often enriched with vegetables, legumes, small fish or eggs and cooking oil. Children liked the taste of enriched porridge. Amount of enriched porridge consumed at each sitting increase among previously fussy-eater children. Meal frequency increased by including fruits as snacks in child’s diet. Grandmothers observed preparation of enriched porridge among the mothers using locally available foods. Grandmothers liked the taste of enriched porridge and not the greenish color of the porridge. Both grandmothers and mothers reported that children were playing independently after consuming enriched porridge and were strong and healthy. These motivated adoption of the practice. Conclusion: Increased knowledge and skill of preparation and utilisation of locally available foods promoted children’s dietary diversification. Children liking the enriched porridge motivated adoption of dietary diversification.

Keywords: behaviour change, complementary feeding, dietary diversification, IYCN

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6409 Local People’s Livelihoods and Coping Strategies in the Wake of a Co-management System in the Campo Ma'an National Park, Cameroon

Authors: Nchanji Yvonne Kiki, Mala William Armand, Nchanji Eileen Bogweh, Ramcilovik-Suominen Sabaheta, Kotilainen Juha

Abstract:

The Campo Ma'an National Park was created as part of an environmental and biodiversity compensation for the Chad-Cameroon Oil Pipeline Project, which was meant to help alleviate poverty and boost the livelihood of rural communities around the area. This paper examines different strategies and coping mechanisms employed by the indigenous people and local communities to deal with the national and internationally driven conservation policies and initiatives in the case of the Campo Ma'an National Park. While most literature on park management/co-management/nature conservation has focused on the negative implications for local peoples’ livelihoods, fewer studies have investigated the strategies of local people to respond to these policies and renegotiate their position in a way that enables them to continue their traditional livelihoods using the existing local knowledge systems. This study contributes to the current literature by zooming into not only the impacts of nature conservation policies but also the local individual and collective strategies and responses to such policies and initiatives. We employ a qualitative research approach using ethnomethodology and a convivial lens to analyze data collected from October to November 2018. We find that conservation policies have worsened some existing livelihoods on the one hand and constrained livelihood improvement of indigenous people and local communities (IPLC) on the other hand. Nonetheless, the IPLC has devised individual and collective coping mechanisms to deal with these conservation interventions and the negative effects they have caused. Upon exploring these mechanisms and their effectiveness, this study proposes a management approach to conservation centered on both people and nature, based on indigenous and local people's knowledge and practices, promoting nature for and by humans and strengthening both livelihood and conservation. We take inspiration from the convivial conservation approach and thinking by Bucher and Fletcher.

Keywords: conservation policies, national park management, indigenous and local people’s experiences, livelihoods, local knowledge, coping strategies, conviviality

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6408 Examples of Techniques and Algorithms Used in Wlan Security

Authors: Vahid Bairami Rad

Abstract:

Wireless communications offer organizations and users many benefits such as portability and flexibility, increased productivity, and lower installation costs. Wireless networks serve as the transport mechanism between devices and among devices and the traditional wired networks (enterprise networks and the internet). Wireless networks are many and diverse but are frequently categorized into three groups based on their coverage range: WWAN, WLAN, and WPAN. WWAN, representing wireless wide area networks, includes wide coverage area technologies such as 2G cellular, Cellular Digital Packet Data (CDPD), Global System for Mobile Communications (GSM), and Mobitex. WLAN, representing wireless local area networks, includes 802.11, Hyper lan, and several others. WPAN, represents wireless personal area network technologies such as Bluetooth and Infrared. The security services are provided largely by the WEP (Wired Equivalent Privacy) protocol to protect link-level data during wireless transmission between clients and access points. That is, WEP does not provide end-to-end security but only for the wireless portion of the connection.

Keywords: wireless lan, wired equivalent privacy, wireless network security, wlan security

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6407 Detection and Quantification of Viable but Not Culturable Vibrio Parahaemolyticus in Frozen Bivalve Molluscs

Authors: Eleonora Di Salvo, Antonio Panebianco, Graziella Ziino

Abstract:

Background: Vibrio parahaemolyticus is a human pathogen that is widely distributed in marine environments. It is frequently isolated from raw seafood, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to acute gastroenteritis. Vibrio spp. has excellent resistance to low temperatures so it can be found in frozen products for a long time. Recently, the viable but non-culturable state (VBNC) of bacteria has attracted great attention, and more than 85 species of bacteria have been demonstrated to be capable of entering this state. VBNC cells cannot grow in conventional culture medium but are viable and maintain metabolic activity, which may constitute an unrecognized source of food contamination and infection. Also V. parahaemolyticus could exist in VBNC state under nutrient starvation or low-temperature conditions. Aim: The aim of the present study was to optimize methods and investigate V. parahaemolyticus VBNC cells and their presence in frozen bivalve molluscs, regularly marketed. Materials and Methods: propidium monoazide (PMA) was integrated with real-time polymerase chain reaction (qPCR) targeting the tl gene to detect and quantify V. parahaemolyticus in the VBNC state. PMA-qPCR resulted highly specific to V. parahaemolyticus with a limit of detection (LOD) of 10-1 log CFU/mL in pure bacterial culture. A standard curve for V. parahaemolyticus cell concentrations was established with the correlation coefficient of 0.9999 at the linear range of 1.0 to 8.0 log CFU/mL. A total of 77 samples of frozen bivalve molluscs (35 mussels; 42 clams) were subsequently subjected to the qualitative (on alkaline phosphate buffer solution) and quantitative research of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose (TCBS) agar (DIFCO) NaCl 2.5%, and incubation at 30°C for 24-48 hours. Real-time PCR was conducted on homogenate samples, in duplicate, with and without propidium monoazide (PMA) dye, and exposed for 45 min under halogen lights (650 W). Total DNA was extracted from cell suspension in homogenate samples according to bolliture protocol. The Real-time PCR was conducted with species-specific primers for V. parahaemolitycus. The RT-PCR was performed in a final volume of 20 µL, containing 10 µL of SYBR Green Mixture (Applied Biosystems), 2 µL of template DNA, 2 µL of each primer (final concentration 0.6 mM), and H2O 4 µL. The qPCR was carried out on CFX96 TouchTM (Bio-Rad, USA). Results: All samples were negative both to the quantitative and qualitative detection of V. parahaemolyticus by the classical culturing technique. The PMA-qPCR let us individuating VBNC V. parahaemolyticus in the 20,78% of the samples evaluated with a value between the Log 10-1 and Log 10-3 CFU/g. Only clams samples were positive for PMA-qPCR detection. Conclusion: The present research is the first evaluating PMA-qPCR assay for detection of VBNC V. parahaemolyticus in bivalve molluscs samples, and the used method was applicable to the rapid control of marketed bivalve molluscs. We strongly recommend to use of PMA-qPCR in order to identify VBNC forms, undetectable by the classic microbiological methods. A precise knowledge of the V.parahaemolyticus in a VBNC form is fundamental for the correct risk assessment not only in bivalve molluscs but also in other seafood.

Keywords: food safety, frozen bivalve molluscs, PMA dye, Real-time PCR, VBNC state, Vibrio parahaemolyticus

Procedia PDF Downloads 128