Search results for: dose rate

Authors: Hadjer Sadoune, Liza Lamini, Scherazade Krim, Amel Djouadi, Rachida Rihani

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Deep neural networks are highly regarded for their accuracy in predicting intricate fermentation processes. Their ability to learn from a large amount of datasets through artificial intelligence makes them particularly effective models. The primary obstacle in improving the performance of these models is to carefully choose the suitable hyperparameters, including the neural network architecture (number of hidden layers and hidden units), activation function, optimizer, learning rate, and other relevant factors. This study predicts biogas production from real wastewater treatment plant data using a sophisticated approach: hybrid Bayesian optimization with a tree-structured Parzen estimator (BO-TPE) for an optimised deep neural network (DNN) model. The plant utilizes an Upflow Anaerobic Sludge Blanket (UASB) digester that treats industrial wastewater from soft drinks and breweries. The digester has a working volume of 1574 m3 and a total volume of 1914 m3. Its internal diameter and height were 19 and 7.14 m, respectively. The data preprocessing was conducted with meticulous attention to preserving data quality while avoiding data reduction. Three normalization techniques were applied to the pre-processed data (MinMaxScaler, RobustScaler and StandardScaler) and compared with the Non-Normalized data. The RobustScaler approach has strong predictive ability for estimating the volume of biogas produced. The highest predicted biogas volume was 2236.105 Nm³/d, with coefficient of determination (R2), mean absolute error (MAE), and root mean square error (RMSE) values of 0.712, 164.610, and 223.429, respectively.

Keywords: anaerobic digestion, biogas production, deep neural network, hybrid bo-tpe, hyperparameters tuning

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Authors: Yi Yang, Jiasong Sun

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Almost every aspect of our daily lives is now intertwined with some degree of human-computer interaction (HCI). HCI courses draw on knowledge from disciplines as diverse as computer science, psychology, design principles, anthropology, and more. Our HCI courses, named the Media and Cognition course, are constantly updated to reflect state-of-the-art technological advancements such as virtual reality, augmented reality, and artificial intelligence-based interactions. For more than a decade, our course has used an interest-based approach to teaching, in which students proactively propose some research-based questions and collaborate with teachers, using course knowledge to explore potential solutions. Semantic communication plays a key role in facilitating understanding and interaction between users and computer systems, ultimately enhancing system usability and user experience. The advancements in AI-generated technology, which have gained significant attention from both academia and industry in recent years, are exemplified by language models like GPT-3 that generate human-like dialogues from given prompts. Our latest version of the Human-Computer Interaction course practices a semantic communication platform based on AI-generated techniques. The purpose of this semantic communication is twofold: to extract and transmit task-specific information while ensuring efficient end-to-end communication with minimal latency. An AI-generated semantic communication platform evaluates the retention of signal sources and converts low-retain ability visual signals into textual prompts. These data are transmitted through AI-generated techniques and reconstructed at the receiving end; on the other hand, visual signals with a high retain ability rate are compressed and transmitted according to their respective regions. The platform and associated research are a testament to our students' growing ability to independently investigate state-of-the-art technologies.

Keywords: human-computer interaction, media and cognition course, semantic communication, retainability, prompts

Procedia PDF Downloads 113

Authors: Hui Liu, Xiaozan Wang, Jiarong Zhong, Ziming Shao

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Introduction: Related research shows that students don’t concentrate on teacher’s speaking in the classroom. The d2 attention test is a time-limited test about selective attention. The d2 attention test can be used to evaluate individual selective attention. Purpose: To use the d2 attention test tool to measure the difference between the attention level of the experimental class and the control class before and after Brain-Break and to explore the effect of Brain-Break in the classroom on students' selective attention. Methods: According to the principle of no difference in pre-test data, two classes in the fourth- grade of Shenzhen Longhua Central Primary School were selected. After 20 minutes of class in the third class in the morning and the third class in the afternoon, about 3-minute Brain-Break intervention was performed in the experimental class for 10 weeks. The normal class in the control class did not intervene. Before and after the experiment, the d2 attention test tool was used to test the attention level of the two-class students. The paired sample t-test and independent sample t-test in SPSS 23.0 was used to test the change in the attention level of the two-class classes around 10 weeks. This article only presents results with significant differences. Results: The independent sample t-test results showed that after ten-week of Brain-Break, the missed errors (E1 t = -2.165 p = 0.042), concentration performance (CP t = 1.866 p = 0.05), and the degree of omissions (Epercent t = -2.375 p = 0.029) in experimental class showed significant differences compared with control class. The students’ error level decreased and the concentration increased. Conclusions: Adding Brain-Break interventions in the classroom can effectively improve the attention level of fourth-grade primary school students to a certain extent, especially can improve the concentration of attention and decrease the error rate in the tasks. The new sport's learning model is worth promoting

Keywords: cultural class, micromotor, attention, D2 test

Procedia PDF Downloads 131

Authors: Mohammed Alassim

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In 2014, a UN survey stated that: "The greatest challenge to the adoption of whole-of government, which fundamentally rests on increased collaboration, is resistance to change among government actors". Change management has experienced both theoretically and practically many transformation over the years. When organizations have to implement radical changes, they have to encounter a plethora of issues which leads to ineffective or inefficient implementation of change in most cases. 70% of change projects fail because of human issues. It has been cited that” most studies still show a 60-70% failure rate for organizational change projects — a statistic that has stayed constant from the 1970’s to the present.”. E-government involves not just technical change but cultural, policy, social and organizational evolution. Managing change and overcoming resistance to change is seen as crucial in the success of E-government projects. Resistance can be from different levels in the organization (top management, middle management or employees at operational levels). There can be many reasons for resistance including fear of change and insecurity, lack of knowledge and absence of commitment from management to implement the change. The purpose of this study is to conduct in-depth research to understand the process of change and to identify the critical factors that have led to resistance from employees at different levels (top management, Middle management and operational employees) during e-government initiatives in the public sector in Saudi Arabia. The study is based on qualitative and empirical research methods conducted in the public sector in the Kingdom of Saudi Arabia. This research will use triangulation in data method (interview, group discussion and document review). This research will contribute significantly to knowledge in this field and will identify the measures that can be taken to reduce resistance to change, Upon analysis recommendations or model will be offered which can enable decision makers in public sector in Saudi Arabia how to plan, implement and evaluate change in e-government initiatives via change management strategy.

Keywords: change management, e-government, managing change, resistance to change

Procedia PDF Downloads 315

Authors: Lulu Liao, Huijing Chen, Yinan Zhao, Hongting Ning, Hui Feng

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Urinary tract infection rate is an important index of care quality in nursing homes. The aim of the study is to understand the nursing assistant's current knowledge and attitudes of urinary incontinence and to explore related stakeholders' viewpoint about urinary incontinence training. This explanatory sequential study used Knowledge, Practice, and Attitude Model (KAP) and Adult Learning Theories, as the conceptual framework. The researchers collected data from 509 nursing assistants in sixteen nursing homes in Hunan province in China. The questionnaire survey was to assess the knowledge and attitude of urinary incontinence of nursing assistants. On the basis of quantitative research and combined with focus group, training demands were identified, which nurse managers should adopt to improve nursing assistants’ professional practice ability in urinary incontinence. Most nursing assistants held the poor knowledge (14.0 ± 4.18) but had positive attitudes (35.5 ± 3.19) toward urinary incontinence. There was a significant positive correlation between urinary incontinence knowledge and nursing assistants' year of work and educational level, urinary incontinence attitude, and education level (p < 0.001). Despite a general awareness of the importance of prevention of urinary tract infections, not all nurse managers fully valued the training in urinary incontinence compared with daily care training. And the nursing assistants required simple education resources to equip them with skills to address problem about urinary incontinence. The variety of learning methods also highlighted the need for educational materials, and nursing assistants had shown a strong interest in online learning. Related education material should be developed to meet the learning need of nurse assistants and provide suitable training method for planned quality improvement in urinary incontinence.

Keywords: mixed methods, nursing assistants, nursing homes, urinary incontinence

Procedia PDF Downloads 136

Authors: Xin Chen, Xiao Yang, Rongrong Han, Lu Chen, Lingling Gao

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Background: Leisure time physical activity (LTPA) benefits both pregnant women and their fetuses. The guidelines recommended that pregnant women should do at least 150 minutes of moderate-intensity aerobic physical activity throughout the week. The aim of this study was to investigate the rate of LTPA participation among Chinese pregnant women and to identify its predictors based on the health belief model. Methods: A cross-sectional study was conducted from June 2019 to September 2019 in Changchun, China. A total of 225 pregnant women aged 18 years or older with no severe physical or mental disease were recruited in the obstetric clinic. Self-administered questionnaires were used to collect data. LTPA was assessed by a pregnant physical activity questionnaire (PPAQ). A revised pregnancy physical activity health belief scale and social-demographic and perinatal characteristics factors were collected and used to predict LTPA participation. Data were analyzed using descriptive statistics and multivariate logistic regression. Results: The participants had a high level of perceived susceptibility, perceived severity, perceived benefits, and action clues, with mean item scores above 3.5. The predictors of LTPA in Chinese pregnant women were pre-pregnancy exercise habits [OR 3.236 (95% CI:1.632, 6.416)], perceived susceptibility score [OR 2.083 (95% CI:1.002, 4.331)], and perceived barriers score [OR 3.113 (95%CI:1.462, 6.626)]. Conclusions: The results of this study will lead to better identification of pregnant women who may not participate in LTPA. Healthcare professionals should be cognizant of issues that may affect LTPA participation among pregnant women, including pre-pregnancy exercise habits, perceived susceptibility, and perceived barriers.

Keywords: pregnancy, health belief model., leisure time physical activity, factors

Procedia PDF Downloads 78

Authors: B. Baba Ahmed, P. Yanogo, B. Djibryl. N. Medas

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Introduction: Polio is a highly contagious viral infection, with children under 5 years of age being the most affected. It is a public health emergency of international concern. Polio surveillance in Mauritania has been ongoing since 1998 and has achieved "polio free" status in 2007. our objective is to analyse a pidemiological surveillance database of poliomyélitis in Mauritania from 2012-2019. Method: A transversal descriptive analysis of poliomyélitis database was carried out in Mauritania from 2012-2019.An exhaustive sampling was done on all suspected polio cases recorded in the database from 2012 -2019. This study used EPI-INFO 7.4 for frequency calculation for qualitative variables, mean and standard deviation for quantitative variables. Results: We found 459 suspected cases of polio over the study period with an average rate of acute non-polio flaccid paralysis of 25.4 cases/100,000 children under 15 years of age. The age group 0-6 years represented 75.2%. Males constituted 50.2%. Females represented 49.78% with a ratio of M/F=1.Among the 422 observations, the average age is 4 years +/- 3.38. The four regions, TIRIS-ZEMMOUR, INCHIRI, TAGANT, NOUACHCHOTT OUEST recorded the lowest percentages of notifications, respectively (3.28%; 3.93%; 4.37%; 4.8%). 99.34% [98.09-99.78] of cases presented acute flaccid paralysis. And 56.77% [52.19-61.23], had limb asymmetry. We showed that 82.93% [79.21-86.10], had fever. we found that 89.5% of suspected polio cases were investigated before 48 hours. And 88.39% of suspected cases had two adequate samples taken 48 hours apart and within 14 days after the onset of symptoms. Only 30.95% of samples arrived at the referral laboratory before 72 hours. Conclusion: This study has shown that Mauritania has achieved the objectives in most of the quantitative performance indicators of polio surveillance. This study has shown a low notification of cases in the northern and central regions of the country. There is a problem with the transport of samples to the laboratory.

Keywords: analysis, data base, Epi-Info, polio

Procedia PDF Downloads 175

Authors: Sajjad Seifi Mofarah, S. K. Sadrnezhaad, Shokooh Moghadam, Javad Tavakoli

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In recent years a new method of combination treatment for cancer has been developed and studied that has led to significant advancements in the field of cancer therapy. Hyperthermia is a traditional therapy that, along with a creation of a medically approved level of heat with the help of an alternating magnetic AC current, results in the destruction of cancer cells by heat. This paper gives details regarding the production of the spherical nanocomposite PVA/γ-Fe2O3 in order to be used for medical purposes such as tumor treatment by hyperthermia. To reach a suitable and evenly distributed temperature, the nanocomposite with core-shell morphology and spherical form within a 100 to 200 nanometer size was created using phase separation emulsion, in which the magnetic nano-particles γ-Fe2O3 with an average particle size of 20 nano-meters and with different percentages of 0.2, 0.4, 0.5, and 0.6 were covered by polyvinyl alcohol. The main concern in hyperthermia and heat treatment is achieving desirable specific absorption rate (SAR) and one of the most critical factors in SAR is particle size. In this project all attempts has been done to reach minimal size and consequently maximum SAR. The morphological analysis of the spherical structure of the nanocomposite PVA/γ-Fe2O3 was achieved by SEM analyses and the study of the chemical bonds created was made possible by FTIR analysis. To investigate the manner of magnetic nanocomposite particle size distribution a DLS experiment was conducted. Moreover, to determine the magnetic behavior of the γ-Fe2O3 particle and the nanocomposite PVA/γ-Fe2O3 in different concentrations a VSM test was conducted. To sum up, creating magnetic nanocomposites with a spherical morphology that would be employed for drug loading opens doors to new approaches in developing nanocomposites that provide efficient heat and a controlled release of drug simultaneously inside the magnetic field, which are among their positive characteristics that could significantly improve the recovery process in patients.

Keywords: nanocomposite, hyperthermia, cancer therapy, drug releasing

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Authors: Nino Chikhladze, Tamar Dochviri, Nato Pitskhelauri, Maia Bitskhinashvili

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Population aging is a key social and demographic trend of the 21st century. Falls represent a prevalent geriatric syndrome that poses significant risks to the health and independence of older adults. The World Health Organization notes a lack of comprehensive data on falls in low- and middle-income countries, complicating the creation of effective prevention programs. To the authors’ best knowledge, no such studies have been conducted in Georgia. The aim of the study is to explore the epidemiology of falls in the elderly population. The hospitalization database of the National Center for Disease Control and Public Health of Georgia was used for the retrospective study. Falls-related injuries were identified using ICD-10 classifications using the class XIX (S and T codes) and class XX for the type of injury (V-Y codes). Statistical data analyses were done using SPSS software version 23.0. The total number of fall-related hospitalizations for individuals aged 65 and older from 2015 to 2021 was 29,697. The study revealed that falls accounted for an average of 63% (ranging from 59% to 66%) of all hospitalizations and 68% (ranging from 65% to 70%) of injury-related hospitalizations during this period. The 69% of all patients were women and 31%-men (Chi2=4482.1, p<0.001). The highest rate of hospitalization was in the age groups 80-84 and 75-79. The probability of fall-related hospitalization was significantly higher in women (p<0.001) compared to men in all age groups except 65-69 years. In the target age group of 65 years and older, the probability of hospitalization increased significantly with an increase in age (p<0.001). The study's results can be leveraged to create evidence-based awareness programs, design targeted multi-domain interventions addressing specific risk factors, and enhance the quality of geriatric healthcare services in Georgia.

Keywords: elderly population, falls, geriatric patients, hospitalization, injuries

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Authors: Abbasali Abouei Mehrizi, Hao Wang

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The thin film region is so important in heat transfer process due to its low thermal resistance. On the other hand, the dynamic contact angle is crucial boundary condition in numerical simulations. While different modeling contains different assumption of the microscopic contact angle, none of them has experimental evidence for their assumption, and the contact line movement mechanism still remains vague. The experimental investigation in complete wetting is more popular than partial wetting, especially in nanoscale resolution when there is sharp variation in thin film profile in partial wetting. In the present study, an experimental investigation of water film morphology near the triple phase contact line during the condensation is performed. The state-of-the-art tapping-mode atomic force microscopy (TM-AFM) was used to get the high-resolution film profile goes down to 2 nm from the contact line. The droplet was put in saturated chamber. The pristine silicon wafer was used as a smooth substrate. The substrate was heated by PI film heater. So the chamber would be over saturated by droplet evaporation. By turning off the heater, water vapor gradually started condensing on the droplet and the droplet advanced. The advancing speed was less than 20 nm/s. The dominant results indicate that in contrast to nonvolatile liquid, the film profile goes down straightly to the surface till 2 nm from the substrate. However, small bending has been observed below 20 nm, occasionally. So, it can be claimed that for the low condensation rate the microscopic contact angle equals to the optically detectable macroscopic contact angle. This result can be used to simplify the heat transfer modeling in partial wetting. The experimental result of the equality of microscopic and macroscopic contact angle can be used as a solid evidence for using this boundary condition in numerical simulation.

Keywords: advancing, condensation, microscopic contact angle, partial wetting

Procedia PDF Downloads 294

Authors: Ghada Esheba, Ghadeer Aldoobi, Salwa Almalk, Abrar Alshareef, Eman Al-khairi, Eman Yaseen

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Background: In Saudi Arabia, ovarian cancer ranked seventh among female population and is the most common female genital tract malignancy after endometrial cancer. A slight increase in the incidence of ovarian cancer was observed from 2001–2008. Makkah, Riyadh, and the eastern region of Saudi Arabia had the highest incidence rate ratio for the number of ovarian cancer cases (1). Differentiating metastatic adenocarcinomas from primary ovarian carcinomas, especially those of endometrioid and mucinous type is clinically significant and a challenge for clinicians and pathologists, yet the distinction has important therapeutic and prognostic implications. Aim: To clarify the most important histopathological criteria to differentiate between primary ovarian surface epithelial tumors especially mucinous and endometrioid subtypes, and metastatic adenocarcinoma and to evaluate the value of a panel of antibodies consisting of CK7, CK20, and CDX-2 in the distinction between primary ovarian surface epithelial tumors and metastatic adenocarcinoma. Material and methods: This study was carried out on 26 cases of primary ovarian surface epithelial neoplasms and 14 cases of metastatic ovarian adenocarcinoma. All cases were studied immunohistochemically using CK7, CK20, and CDX-2. Results: All cases of primary ovarian adenocarcinoma were positive for CK7. 25% and 58% of mucinous borderline mucinous tumor and mucinous carcinoma respectively were positive for CK20. Only 42% of mucinous carcinoma were positive for CDX-2. All cases of endometrioid carcinomas were negative for both CK20 and CDX-2. All cases of metastatic adenocarcinoma from the colon were negative for CK7 and positive for CK20 and CDX-2. Conclusions: CK7 is an important positive marker for primary ovarian tumors, while CK20 and CDX-2 are useful markers for colorectal carcinoma metastatic to the ovary. Caution should be taken as primary ovarian mucinous tumors may stain positive for CK20, CDX-2, or both, however, they usually exhibit a focal pattern of reactivity.

Keywords: adenoma, endometrioid, malignancy, ovarian

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Authors: Abdulaziz Abdulrahman Albahar

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This paper aims to answer the question of: is oil as a natural resource with all the wealth that it brings an economic burden? And how can resource curse be mitigated in such oil dependent nations? The case of Kuwait will be used as an example. The paper begins with an introduction of the resource curse and the Kuwaiti economy in general. Then there is an attempt to see that does the curse exist in the case for Kuwait. Furthermore, in the analysis section, an exploration on how the economy is dependent on oil and how oil is more of a burden if there is mismanagement is conducted. Later on, in answering on how to mitigate the problem of a resource curse, the case of Norway is explored. In concluding the paper, the results do show that oil rentals affects the Kuwaiti economy via 2 main channels, these are government spending that are mainly financed via oil rentals and exportation of oil based products. The surprising result was that government spending had a negative impact on GDP (gross domestic product) growth when oil rentals where instrumented on government expenditure, this is due to the issue of rent seeking in which government spending in Kuwait is financing things such as stimulus packages and raising the nominal wages. Yet, when comparing the magnitude of both oil exportation and government spending, the latter has a stronger effect on the GDP (gross domestic product) growth than the former. A resource curse doesn’t seem to exist in the case of Kuwait however, the characteristics of a curse do show in the form of rent seeking in the political sphere, the disruption of the traditional sectors like that of pearl trade and fishing markets. Yet, a curse doesn’t show due to the fact that the currency of the nation is very stable and hasn’t experienced any appreciation because of the fixed exchange rate system. Moreover, even if we can’t say that a curse exists, it is clear to see that the Kuwaiti economy is heading towards one. Whether or not it faces a resource curse will be based on how judicious the nation will be in exploiting their sovereign wealth fund and implementing diversification strategies to be less oil dependent like the vision “New Kuwait-2035” which has been underway since 2017.

Keywords: economic development, Kuwait, oil curse, dutch disease

Procedia PDF Downloads 75

Authors: Shirin Ghatrehsamani

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Higher efficiency and lower environmental impact are the consequence of using advanced technology in farming. They also help to decrease yield variability by diminishing weather variability impact, optimizing nutrient and pest management as well as reducing competition from weeds. A better understanding of the pros and cons of applying technology and finding the main reason for preventing the utilization of the technology has a significant impact on developing technology adoption among farmers and producers in the digital agriculture era. The results from two surveys carried out in 2019 and 2021 were used to investigate whether the crop types had an impact on the willingness to utilize technology on the farms. The main focus of the questionnaire was on utilizing precision agriculture (PA) technologies among farmers in some parts of the united states. Collected data was analyzed to determine the practical application of various technologies. The survey results showed more similarities in the main reason not to use PA between the two crop types, but the present application of using technology in specialty crops is generally five times larger than in row crops. GPS receiver applications were reported similar for both types of crops. Lack of knowledge and high cost of data handling were cited as the main problems. The most significant difference was among using variable rate technology, which was 43% for specialty crops while was reported 0% for row crops. Pest scouting and mapping were commonly used for specialty crops, while they were rarely applied for row crops. Survey respondents found yield mapping, soil sampling map, and irrigation scheduling were more valuable for specialty crops than row crops in management decisions. About 50% of the respondents would like to share the PA data in both types of crops. Almost 50 % of respondents got their PA information from retailers in both categories, and as the second source, using extension agents were more common in specialty crops than row crops.

Keywords: precision agriculture, smart farming, digital agriculture, technology adoption

Procedia PDF Downloads 113

Authors: Jegede Kehinde Jacob, Ola Akeem Bayonle, Adewale Yemi Yekeen

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The rapid rate of urban growth in most developing countries of the world in recent times is alarming. Mass movement of people from rural areas to the urban centres, the consequence of the uncontrolled rapid urbanisation resulting to many un-conforming environmental challenges such as inadequate infrastructure, land, water and air pollution, poor environmental sanitation, poor and inadequate housing, urban degradation, sprawl and slums, urban violence, crime, robbery and prostitution as well as many other social vices that make the cities unsustainable. The resultant effects of all these are abysmal failure in the management of cities on the part of the governing authorities and other relevant stakeholders as well as unconducive and unwholesome condition of living of the people. This paper attempts to examine holistically the issue of environmental planning management (EPM) process development and management concept with a view for dynamic and interactive approach for various stakeholders as partners in achieving sustainable cities of our dream. The areas of discussion including conceptual and contextual issues, sustainable cities concept, good urban governance including literature review. The paper goes further to examine opportunities and challenges of built environment generally, the nature and context of environmental problems in particular, the role and duties of environmental planning and management (EPM) process in sustainable urban development. The paper further reviewed briefly the various levels of institutionalisation of EPM process with a typical case study of sustainable Ibadan project (SIP). The paper concludes with a list of recommendations to ensure effective and lasting solutions to cities problems through initiation of EPM process achievable in a sustainable manner.

Keywords: built environment, environmental planning, sustainable cities, sustainable development, urbanization

Procedia PDF Downloads 262

Authors: Kara Rosenblatt, Kevin Badgett, James Eldridge

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There is a preponderance of information regarding the overall effectiveness of charter schools and their ability to increase academic achievement compared to traditional district schools. Most research on the topic is focused on comparing long and short-term outcomes, academic achievement in mathematics and reading, and locale (i.e., urban, v. Rural). While the lingering unanswered questions regarding effectiveness continue to loom for school leaders, data on charter schools suggests that enrollment increases by 10% annually and that charter schools educate more than 2 million U.S. students across 40 states each year. Given the increasing share of U.S. students educated in charter schools, it is important to better understand possible differences in student achievement defined in multiple ways for students in charter schools and for those in Independent School District (ISD) settings in the state of Texas. Data were retrieved from the Texas Education Agency’s (TEA) repository that includes data organized annually and available on the TEA website. Specific data points and definitions of achievement were based on characterizations of achievement found in the relevant literature. Specific data points include but were not limited to graduation rate, student performance on standardized testing, and teacher-related factors such as experience and longevity in the district. Initial findings indicate some similarities with the current literature on long-term student achievement in English/Language Arts; however, the findings differ substantially from other recent research related to long-term student achievement in social studies. There are a number of interesting findings also related to differences between achievement for students in charters and ISDs and within different types of charter schools in Texas. In addition to findings, implications for leadership in different settings will be explored.

Keywords: charter schools, ISDs, student achievement, implications for PK-12 school leadership

Procedia PDF Downloads 124

Authors: Sarah Gaßmeyer, Nadine Hülsemann, Raphael Stoll, Kenji Miyamoto, Robert Kourist

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Enzymatic racemization allows the smooth interconversion of stereocenters under very mild reaction conditions. Racemases find frequent applications in deracemization and dynamic kinetic resolutions. Arylmalonate decarboxylase (AMDase) from Bordetella Bronchiseptica has high structural similarity to amino acid racemases. These cofactor-free racemases are able to break chemically strong CH-bonds under mild conditions. The racemase-like catalytic machinery of mutant G74C conveys it a unique activity in the racemisation of pharmacologically relevant derivates of 2-phenylpropionic acid (profenes), which makes AMDase G74C an interesting object for the mechanistic investigation of cofactor-independent racemases. Structure-guided protein engineering achieved a variant of this unique racemase with 40-fold increased activity in the racemisation of several arylaliphatic carboxylic acids. By saturation–transfer–difference NMR spectroscopy (STD-NMR), substrate binding during catalysis was investigated. All atoms of the substrate showed interactions with the enzyme. STD-NMR measurements revealed distinct nuclear Overhauser effects in experiments with and without molecular conversion. The spectroscopic analysis led to the identification of several amino acid residues whose variation increased the activity of G74C. While single-amino acid exchanges increased the activity moderately, structure-guided saturation mutagenesis yielded a quadruple mutant with a 40 times higher reaction rate. This study presents STD-NMR as versatile tool for the analysis of enzyme-substrate interactions in catalytically competent systems and for the guidance of protein engineering.

Keywords: racemase, rational protein design, STD-NMR, structure guided saturation mutagenesis

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Authors: Marisa Chrysochoou, James Mahoney, Kay Wille

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Pyrrhotite is an iron-sulfide mineral which releases sulfuric acid when exposed to water and oxygen. The presence of this mineral in concrete foundations across Connecticut and Massachusetts in the US is causing in some cases premature failure. This has resulted in a devastating crisis for all parties affected by this type of failure which can take up to 15-25 years before internal damage becomes visible on the surface. This study shares laboratory results aimed to investigate the fundamental mechanisms of pyrrhotite reaction and to further the understanding of its deterioration kinetics within concrete. This includes the following analyses: total sulfur, wavelength dispersive X-ray fluorescence, expansion, reaction rate combined with ion-chromatography, as well as damage evolution using electro-chemical acceleration. This information is coupled to a statistical analysis of over 150 analyzed concrete foundations. Those samples were obtained and process using a developed and validated sampling method that is minimally invasive to the foundation in use, provides representative samples of the concrete matrix across the entire foundation, and is time and cost-efficient. The processed samples were then analyzed using a developed modular testing method based on total sulfur and wavelength dispersive X-ray fluorescence analysis to quantify the amount of pyrrhotite. As part of the statistical analysis the results were grouped into the following three categories: no damage observed and no pyrrhotite detected, no damage observed and pyrrhotite detected and damaged observed and pyrrhotite detected. As expected, a strong correlation between amount of pyrrhotite, age of the concrete and damage is observed. Information from the laboratory investigation and from the statistical analysis of field samples will aid in forming a scientific basis to support the decision process towards sustainable financial and administrative solutions by state and local stakeholders.

Keywords: concrete, pyrrhotite, risk of failure, statistical analysis

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Authors: Kalaashini Ramachandran, Maznah Dahlui, Nik Daliana Nik Farid

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WHO recommends all babies to be exclusively breastfed and donor milk is the next best alternative in the absence of mother’s own milk. The establishment of a human milk bank (HMB) is still being debated due to religious concerns in Malaysia leading to informal milk sharing practices, but little is known on the knowledge, attitude and perception of women towards HMB and its benefits. This study hypothesizes that there is no association between knowledge and attitude and the acceptance towards the establishment of human milk bank among Malaysian women and healthcare providers. The aim of this study is to determine the drivers and barriers among Malaysian towards the acceptance of an HMB. A cross-sectional study with 367 participants was enrolled within a period of 3 months to answer an online self-administered questionnaire. Data on sociodemographic, knowledge on breastfeeding benefits, knowledge and attitude on HMB and its specific issues were analyzed in terms of frequency and then proceed to multiple logistic regression. Majority of the respondents are of Islamis religion (73.3%), have succeesfully completed their tertiary education (82.8%), and are employed (70.8%). Only 55.9% of respondents have heard of an HMB stating internet as their main source of information but a higher prevalence is agreeable to the establishment of a human milk bank (67.8%). Most respondents have a good score on knowledge of breastfeeding benefits and on HMB specific issues (70% and 54.2% respectively) while 63.8% of them have a positive attitude towards HMB. In the multivariate analysis, mothers with a good score on general knowledge of breastfeeding (AOR: 1.715) were more likely to accept the establishment of an HMB while Islamic religion was negatively associated with its establishment (AOR:0.113). This study has found a high prevalence rate of mothers who are willing to accept the establishment of an HMB. This action can be potentially shaped by educating mothers on the benefits of breastfeeding as well as addressing their religious concerns so the establishment of a religiously abiding HMB in Malaysia may be accepted without compromising their belief or the health benefit of donor milk.

Keywords: acceptability, attitude, human milk bank, knowledge

Procedia PDF Downloads 101

Authors: Nadir Atayev, Mehman Hasanov

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Future spacecraft will require a structural change in the way data is transmitted due to the increase in the volume of data required for space communication. Current radio frequency communication systems are already facing a bottleneck in the volume of data sent to the ground segment due to their technological and regulatory characteristics. To overcome these issues, free space optics communication plays an important role in the integrated terrestrial space network due to its advantages such as significantly improved data rate compared to traditional RF technology, low cost, improved security, and inter-satellite free space communication, as well as uses a laser beam, which is an optical signal carrier to establish satellite-ground & ground-to-satellite links. In this approach, there is a need for high-speed and energy-efficient systems as a base platform for sending high-volume video & audio data. Nano Satellite and its branch CubeSat platforms have more technical functionality than large satellites, wheres cover an important part of the space sector, with their Low-Earth-Orbit application area with low-cost design and technical functionality for building networks using different communication topologies. Along the research theme developed in this regard, the output parameter indicators for the FSO of the optical communication transceiver subsystem on the existing CubeSat platforms, and in the direction of improving the mentioned parameters of this communication methodology, 3D optical switch and laser beam controlled optical transponder with 2U CubeSat structural subsystems and application in the Low Earth Orbit satellite network topology, as well as its functional performance and structural parameters, has been studied accordingly.

Keywords: cubesat, free space optics, nano satellite, optical laser communication.

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Authors: Lubna M. Zakaryia Mahmoud, Mohammed A. Dawood, Doaa A. Heiba

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A prospective double-blind placebo controlled trial carried out on 60 children known to be asthmatic who presented to the emergency department at Alexandria University of Children’s Hospital at El-Shatby with acute asthma exacerbations to assess the efficacy of adding inhaled magnesium sulfate to β-agonist, compared with β-agonist in saline, in the management of acute asthma exacerbations in children. The participants in the study were divided in two groups; Group A (study group) received inhaled salbutamol solution (0.15 ml/kg) plus isotonic magnesium sulfate 2 ml in a nebulizer chamber. Group B (control group): received nebulized salbutamol solution (0.15 ml/kg) diluted with placebo (2 ml normal saline). Both groups received inhaled solution every 20 minutes that was repeated for three doses. They were evaluated using the Pediatric Asthma Severity Score (PASS), oxygen saturation using portable pulse oximetry and peak expiratory flow rate using a portable peak expiratory flow meter at initially recorded as zero-minute assessment and every 20 minutes from the end of each nebulization (nebulization lasts 5-10 minutes) recorded as 20, 40 and 60-minute assessments. Regarding PASS, comparison showed non-significant difference with p-value 0.463, 0.472, 0.0766 at 20, 40 and 60 minutes. Regarding oxygen saturation, improvement was more significant towards group A starting from 40 min with significant p-value=0.000. At 60 min p-value=0.000. Although mean PEFR significantly improved from zero-min in both groups; however, improvement was more significant in group A with significant p-value = 0.015, 0.001, 0.001 at 20 min, 40 min and 60 min, respectively. The conclusion this study suggests is that inhaled magnesium sulfate is an efficient add on drug to standard β- agonist inhalation used in the treatment of moderate to severe asthma exacerbations.

Keywords: nebulized, magnesium sulfate, acute asthma , pediatric

Procedia PDF Downloads 181

Authors: K. Jankowski, M. Pawlikowski, A. Makuch, K. Skalski

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Depth-sensing indentation (DSI) or nanoindentation is becoming a more and more popular method of measuring mechanical properties of various materials and tissues at a micro-scale. This technique allows measurements without complicated sample preparation procedures which makes this method very useful. As a result of measurement force and displacement of the intender are obtained. It is also possible to determine three measures of hardness i.e. Martens hardness (HM), nanohardness (HIT), Vickers hardness (HV) and Young modulus EIT. In this work trabecular bone mechanical properties were investigated. The bone samples were harvested from human femoral heads during hip replacement surgery. Patients were of different age, sexes and stages of tissue degeneration caused by osteoarthritis. The specimens were divided into three groups. Each group contained samples harvested from patients of different range of age. All samples were investigated with the same measurement conditions. The maximum load was Pmax=500 mN and the loading rate was 500 mN/min. The tests were held without hold at the peak force. The tests were conducted with indenter Vickers tip and spherical tip of the diameter 0.2 mm. Each trabecular bone sample was tested 7 times in a close area of the same trabecula. The measured loading P as a function of indentation depth allowed to obtain hysteresis loop and HM, HIT, HV, EIT. Results for arbitrarily chosen sample are HM=289.95 ± 42.31 MPa, HIT=430.75 ± 45.37 MPa, HV=40.66 ± 4.28 Vickers, EIT=7.37 ± 1.84 GPa for Vickers tip and HM=115.19 ± 15.03 MPa, HIT=165.80 ± 19.30 MPa, HV=16.90 ± 1.97 Vickers, EIT=5.30 ± 1.31 GPa for spherical tip. Results of nanoindentation tests show that this method is very useful and is perfect for obtaining mechanical properties of trabecular bone. Estimated values of elastic modulus are similar. The differences between hardness are significant but it is a result of using two different types of tips. However, it has to be emphasised that the differences in the values of elastic modulus and hardness result from different testing protocols, anisotropy and asymmetry of the micro-samples and the hydration of bone.

Keywords: human bone, mechanical properties, nano hardness nanoindentation, trabecular bone

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Authors: Ivan Zupan, Tomislav Saric, Suzana Tkalcic

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IMUNO-2865® is a commercially available, β–glucan based, natural hemicellulose compound with proven immunostimulative properties in people, domestic and some aquatic animals. During the experimental feeding trial with IMUNO-2865® in juvenile wild-caught chub under laboratory conditions, supplementation resulted in overall higher growth performance for all experimental groups regardless of the concentration of the added compound. The maximum, 5% concentration of the supplement, resulted in highest weight gain and calculated specific growth rate. In sea bream, as economically most important species in the Mediterranean aquaculture, significant increases in numbers of monocytes and heterophils were observed in the group supplemented with 2.5 % of IMUNO-2865® in the feed. An overall increase of erythrocytes was noted by the end of the experiment, although with variable distribution among groups. Blood Ca++ levels, total proteins, and total NH₃ were significantly higher after 60 days of feeding in all treatment groups compared to the control and remained elevated in the treated group following the secession of supplementation. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum paraoxonase PON1 (U/L) showed similar trends. All these parameters are playing a significant role in either oxygen supplementation of tissues, or anabolic and catabolic processes that on molecular levels contribute to the overall health and immune-building capacity of cells and tissues. The complete lack of mortality in sea bream and presented increases in cellular, biochemical and oxidative stress parameters in the blood suggest that the IMUNO-2865® represents a safe dietary supplement for in aquaculture, with an overall positive and potentially immunostimulative effect on farmed fish.

Keywords: IMUNO-2865®, β–glucans, Mediterranean aquaculture, fish imunnostimulans

Procedia PDF Downloads 141

Authors: Andres Diaz Garcia

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The successful technological development of any bioproduct, including those of the biostimulant type, requires to adequately completion of a series of stages allied to different disciplines that are related to microbiological, engineering, pharmaceutical chemistry, legal and market components, among others. Engineering as a discipline has a key contribution in different aspects of fermentation processes such as the design and optimization of culture media, the standardization of operating conditions within the bioreactor and the scaling of the production process of the active ingredient that it will be used in unit operations downstream. However, all aspects mentioned must take into account many biological factors of the microorganism such as the growth rate, the level of assimilation to various organic and inorganic sources and the mechanisms of action associated with its biological activity. This paper focuses on the practical experience within the Colombian Corporation for Agricultural Research (Agrosavia), which led to the development of a biostimulant bioproduct based on native rhizobacteria Bacillus amyloliquefaciens, oriented mainly to plant growth promotion in cape gooseberry nurseries and fruit crops in Colombia, and the challenges that were overcome from the expertise in the area of engineering. Through the application of strategies and engineering tools, a culture medium was optimized to obtain concentrations higher than 1E09 CFU (colony form units)/ml in liquid fermentation, the process of biomass production was standardized and a scale-up strategy was generated based on geometric (H/D of bioreactor relationships), and operational criteria based on a minimum dissolved oxygen concentration and that took into account the differences in the capacity of control of the process in the laboratory and pilot scales. Currently, the bioproduct obtained through this technological process is in stages of registration in Colombia for cape gooseberry fruits for export.

Keywords: biochemical engineering, liquid fermentation, plant growth promoting, scale-up process

Procedia PDF Downloads 111

Authors: Sekena H. Abd El-Aziem, Heba A. M. Abd El-Kader, Sally S. Alam, Othman E. Othman

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Growth hormone (GH) is an anabolic hormone synthesized and secreted by the somatotroph cells of the anterior lobe of the pituitary gland in a circadian and pulsatile manner, the pattern of which plays an important role in postnatal longitudinal growth and development, tissue growth, lactation, reproduction as well as protein, lipid and carbohydrate metabolism. The aim of this study was to detect the genetic polymorphism of GH gene in five camel breeds reared in Egypt; Sudany, Somali, Mowaled, Maghrabi and Falahy, using PCR-RFLP technique. Also this work aimed to identify the single nucleotide polymorphism between different genotypes detected in these camel breeds. The amplified fragment of camel GH at 613-bp was digested with the restriction enzyme MspI and the result revealed the presence of three different genotypes; CC, CT and TT in tested breeds and significant differences were recorded in the genotype frequencies between these camel breeds. The result showed that the Maghrabi breed that is classified as a dual purpose camels had higher frequency for allele C (0.75) than those in the other tested four breeds. The sequence analysis declared the presence of a SNP (C→T) at position 264 in the amplified fragment which is responsible for the destruction of the restriction site C^CGG and consequently the appearance of two different alleles C and T. The nucleotide sequences of camel GH alleles T and C were submitted to nucleotide sequences database NCBI/Bankit/GenBank and have accession numbers: KP143517 and KP143518, respectively. It is concluded that only one SNP C→T was detected in GH gene among the five tested camel breeds reared in Egypt and this nucleotide substitution can be used as a marker for the genetic biodiversity between camel breeds reared in Egypt. Also, due to the possible association between allele C and higher growth rate, we can used it in MAS for camels and enter the camels possess this allele in breeding program as a way for enhancement of growth trait in camel breeds reared in Egypt.

Keywords: camel breeds in Egypt, GH, PCR-RFLP, SNPs

Procedia PDF Downloads 461

Authors: Ali Javaid, Rizwan Latif, Syed Adnan Qasim, Imran Shafi

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To control combustion inside a direct injection diesel engine, fuel atomization is the best tool. Controlling combustion helps in reducing emissions and improves efficiency. Cavitation is one of the most important factors that significantly affect the nature of spray before it injects into combustion chamber. Typical fuel injector nozzles are small and operate at a very high pressure, which limits the study of internal nozzle behavior especially in case of diesel engine. Simulating cavitation in a fuel injector will help in understanding the phenomenon and will assist in further development. There is a parametric variation between high speed and high torque low speed diesel engines. The objective of this study is to simulate internal spray characteristics for a low speed high torque diesel engine. In-nozzle cavitation has strong effects on the parameters e.g. mass flow rate, fuel velocity, and momentum flux of fuel that is to be injected into the combustion chamber. The external spray dynamics and subsequently the air – fuel mixing depends on a lot of the parameters of fuel injecting the nozzle. The approach used to model turbulence induced in – nozzle cavitation for high-torque low-speed diesel engine, is homogeneous equilibrium model. The governing equations were modeled using Matlab. Complete Model in question was extensively evaluated by performing 3-D time-dependent simulations on Open FOAM, which is an open source flow solver and implemented in CFD (Computational Fluid Dynamics). Results thus obtained will be analyzed for better evaporation in the near-nozzle region. The proposed analyses will further help in better engine efficiency, low emission, and improved fuel economy.

Keywords: cavitation, HEM model, nozzle flow, open foam, turbulence

Procedia PDF Downloads 289

Authors: Ibrahim Umar, S Chindo, A Rajab

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Background: TB remains a disease of public health concern in Nigeria with an estimated incidence rate of 219/100,000. Kano has the second highest TB burden in Nigeria and is the leading state with the highest consistent yearly TB notification. House-to-house (H2H) active case search in the community was found to have major contribution to the total TB notification in the state. Aims and Objective: To showcase the impact of H2H community active TB case search (ACF) to yearly TB notification in Kano State, Northern Nigeria from 2012 to 2022. Methodology: This is a retrospective descriptive study based on the analysis of data collected during the routine quarterly and yearly TB data collected in the state. Data was analyzed using the Power BI with statistical alpha level of significance <0.05. Results: Between 2012 and 2013 there was no House-to-house active TB case search in Nigeria and Kano had zero contribution to TB notification from the community in those years. However, in 2014 with the introduction of H2H Active TB Case Search Kano notified 6,014 TB cases out of which 113 came from the community ACF that translated to 2% contribution to total TB notification. From 2014 to 2022 there was progressive increase in community contribution to TB case notification from 113 out of 6,014 total TB patients notified (2012) to 11,799 out of 26,371 TB patients notified (2022) in Kano State. This translated to 45% increase in community contribution to total TB case notification. Discussion: Remarkable increase in community contribution to total TB case notification in Kano State was achieved in 2022 with 11,799 TB cases notified from the community Active TB case search to the total of 26,731 TB cases notified in Kano State, Nigeria. Conclusion: in research has shown that Community-based H2H Active TB Case Search through Community TB Workers (CTWs) is an excellent strategy in finding the missing TB cases towards Ending TB in the world.

Keywords: tuberculosis(TB), active case search (ACF), house-to-house (H2H), community TB workers (CTWs)

Procedia PDF Downloads 88

Authors: Mohammad Aladwan, Adelia Skripova

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Bioremediation aspects of crude oil-polluted fields can be achieved by isolation and identification of bacterial species from oil-contaminated soil in order to choose the most active isolates and increase the strength of others. In this study, oil-degrading bacteria were isolated and identified from oil-contaminated soil samples in northeastern Jordan. The bacterial growth count (CFU/g) was between 1.06×10⁵ and 0.75×10⁹. Eighty-two bacterial isolates were characterized by their morphology and biochemical tests. The identified bacterial genera included: Klebsiella, Staphylococcus, Citrobacter, Lactobacillus, Alcaligenes, Pseudomonas, Hafnia, Micrococcus, Rhodococcus, Serratia, Enterobacter, Bacillus, Salmonella, Mycobacterium, Corynebacterium, and Acetobacter. Molecular identification of a universal primer 16S rDNA gene was used to identify four bacterial isolates: Microbacterium esteraromaticum strain L20, Pseudomonas stutzeri strain 13636M, Klebsilla pneumoniae, and uncultured Klebsilla sp., known as new strains. Our results indicate that their specific oil-degrading bacteria isolates might have a high strength of oil degradation from oil-contaminated sites. Staphylococcus intermedius (75%), Corynebacterium xerosis (75%), and Pseudomonas fluorescens (50%) showed a high growth rate on different types of hydrocarbons, such as crude oil, toluene, naphthalene, and hexane. In addition, monooxygenase and catechol 2,3-dioxygenase were detected in 17 bacterial isolates, indicating their superior hydrocarbon degradation potential. Total petroleum hydrocarbons were analyzed using gas chromatography for soil samples. Soil samples M5, M7, and M8 showed the highest levels (43,645, 47,805, and 45,991 ppm, respectively), and M4 had the lowest level (7,514 ppm). All soil samples were analyzed for heavy metal contamination (Cu, Cd, Mn, Zn, and Pb). Site M7 contains the highest levels of Cu, Mn, and Pb, while Site M8 contains the highest levels of Mn and Zn. In the future, these isolates of bacteria can be used for the cleanup of oil-contaminated soil.

Keywords: bioremediation, 16S rDNA gene, oil-degrading bacteria, hydrocarbons

Procedia PDF Downloads 123

Authors: M. G. Ghanem, A. M. M. A. Allam, Diaa E. Fawzy, Mehmet Faruk Cengiz

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This paper is devoted to the design and implementation of a smartwatch textile antenna for Wi-Fi bio-medical applications in millimetric wave bands. The antenna is implemented on a leather textile-based substrate to be embedded in a smartwatch. It enables the watch to pick Wi-Fi signals without the need to be connected to a mobile through Bluetooth. It operates at 60 GHz or WiGig (Wireless Gigabit Alliance) band with a wide band for higher rate applications. It also could be implemented over many stratified layers of the body organisms to be used in the diagnosis of many diseases like diabetes and cancer. The structure is designed and simulated using CST (Studio Suite) program. The wearable patch antenna has an octagon shape, and it is implemented on leather material that acts as a flexible substrate with a size of 5.632 x 6.4 x 2 mm3, a relative permittivity of 2.95, and a loss tangent of 0.006. The feeding is carried out using differential feed (discrete port in CST). The work provides five antenna implementations; antenna without ground, a ground is added at the back of the antenna in order to increase the antenna gain, the substrate dimensions are increased to 15 x 30 mm2 to resemble the real hand watch size, layers of skin and fat are added under the ground of the antenna to study the effect of human body tissues human on the antenna performance. Finally, the whole structure is bent. It is found that the antenna can achieve a simulated peak realized gain in dB of 5.68, 7.28, 6.15, 3.03, and 4.37 for antenna without ground, antenna with the ground, antenna with larger substrate dimensions, antenna with skin and fat, and bent structure, respectively. The antenna with ground exhibits high gain; while adding the human organisms absorption, the gain is degraded because of human absorption. The bent structure contributes to higher gain.

Keywords: bio medical engineering, millimetric wave, smart watch, textile antennas, Wi-Fi

Procedia PDF Downloads 119

Authors: Firsan Nova

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The average Indonesian watches 5.5 hours of TV a day. With a population of 242 million people and a Free-to-Air (FTA) TV penetration rate of 56%, that equates to 745 million hours of television watched each day. With such potential, it is no wonder that many companies are now attempting to get into the Pay TV market. Research firm Media Partner Asia has forecast in its study that the number of Indonesian pay-television subscribers will climb from 2.4 million in 2012 to 8.7 million by 2020, with penetration scaling up from 7 percent to 21 percent. Key drivers of market growth, the study says, include macro trends built around higher disposable income and a rising middle class, with leading players continuing to invest significantly in sales, distribution and content. New entrants, in the meantime, will boost overall prospects. This study aims to examine and analyze the effect of Market Attractiveness and the Core Competence on Value Creation and Competitive Advantage and its impact to Business Performance in the pay TV industry in Indonesia. The study using strategic management science approach with the census method in which all members of the population are as sample. Verification method is used to examine the relationship between variables. The unit of analysis in this research is all Indonesian Pay TV business units totaling 19 business units. The unit of observation is the director and managers of each business unit. Hypothesis testing is performed by using statistical Partial Least Square (PLS). The conclusion of the study shows that the market attractiveness affects business performance through value creation and competitive advantage. The appropriate value creation comes from the company ability to optimize its core competence and exploit market attractiveness. Value creation affects competitive advantage. The competitive advantage can be determined based on the company's ability to create value for customers and the competitive advantage has an impact on business performance.

Keywords: market attractiveness, core competence, value creation, competitive advantage, business performance

Procedia PDF Downloads 346

Authors: Mohammad K. Parvez, Mohammed S. Al-Dosari

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Hepatitis E virus (HEV) is an emerging RNA virus that causes acute and chronic liver disease with a global mortality rate of about 2%. Despite milestone developments in understanding of HEV biology, there is still lack of a robust culture system or animal model. Therefore, in a novel approach, two recombinant-baculoviruses (vBac-ORF2 and vBac-ORF3) that could overexpress HEV ORF2 (structural/capsid) and ORF3 (nonstructural/regulatory) proteins, respectively were constructed. The established HEV-SAR55 (genotype 1) replicon that contained GFP gene, in place of ORF2/ORF3 sequences was in vitro transcribed, and GFP production in RNA transfected S10-3 cells was scored by FACS. Enhanced infectivity, if any, of nascent virions produced by exogenously-supplied ORF2 and viral RNA by co-expression of ORF3 was tested on naïve HepG2 cells. Co-transduction with vBac-ORF2/vBac-ORF3 (108 pfu/microL) produced high amounts of native ORF2/ORF3 in approximately 60% of S10-3 cells, determined by immunofluorescence microscopy and Western analysis. FACS analysis showed about 9% GFP positivity of S10-3 cells on day6 post-transfection (i.e, day5 post-transduction). Further, FACS scoring indicated that lysates from S10-3 cultures receiving the RNA plus vBac-ORF2 were capable of producing HEV particles with about 4% infectivity in HepG2 cells. However, lysates of cultures co-transduced with vBac-ORF3, were found to further enhance virion infectivity by approximately 17%. This supported a previously proposed role of ORF3 as a minor-structural protein in HEV virion assembly and infectivity. In conclusion, the present model for efficient genomic RNA packaging and production of infectious virions could be a valuable tool to study various aspects of HEV molecular biology, in vitro.

Keywords: chronic liver disease, hepatitis E virus, ORF2, ORF3, replicon

Procedia PDF Downloads 254