Search results for: genetic profile
Commenced in January 2007
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Edition: International
Paper Count: 3545

Search results for: genetic profile

155 Embryonic Aneuploidy – Morphokinetic Behaviors as a Potential Diagnostic Biomarker

Authors: Banafsheh Nikmehr, Mohsen Bahrami, Yueqiang Song, Anuradha Koduru, Ayse K. Vuruskan, Hongkun Lu, Mallory Pitts, Tolga B. Mesen, Tamer M. Yalcinkaya

Abstract:

The number of people who receive in vitro fertilization (IVF) treatment has increased on a startling trajectory over the past two decades. Despite advances in this field, particularly the introduction of intracytoplasmic sperm injection (ICSI) and the preimplantation genetic screening (PGS), the IVF success remains low. A major factor contributing to IVF failure is embryonic aneuploidy (abnormal chromosome content), which often results in miscarriage and birth defects. Although PGS is often used as the standard diagnostic tool to identify aneuploid embryos, it is an invasive approach that could affect the embryo development, and yet inaccessible to many patients due its high costs. As such, there is a clear need for a non-invasive cost-effective approach to identify euploid embryos for single embryo transfer (SET). The reported differences between morphokinetic behaviors of aneuploid and euploid embryos has shown promise to address this need. However, current literature is inconclusive and further research is urgently needed to translate current findings into clinical diagnostics. In this ongoing study, we found significant differences between morphokinetic behaviors of euploid and aneuploid embryos that provides important insights and reaffirms the promise of such behaviors for developing non-invasive methodologies. Methodology—A total of 242 embryos (euploid: 149, aneuploid: 93) from 74 patients who underwent IVF treatment in Carolinas Fertility Clinics in Winston-Salem, NC, were analyzed. All embryos were incubated in an EmbryoScope incubator. The patients were randomly selected from January 2019 to June 2021 with most patients having both euploid and aneuploid embryos. All embryos reached the blastocyst stage and had known PGS outcomes. The ploidy assessment was done by a third-party testing laboratory on day 5-7 embryo biopsies. The morphokinetic variables of each embryo were measured by the EmbryoViewer software (Uniesense FertiliTech) on time-lapse images using 7 focal depths. We compared the time to: pronuclei fading (tPNf), division to 2,3,…,9 cells (t2, t3,…,t9), start of embryo compaction (tSC), Morula formation (tM), start of blastocyst formation (tSC), blastocyst formation (tB), and blastocyst expansion (tEB), as well as intervals between them (e.g., c23 = t3 – t2). We used a mixed regression method for our statistical analyses to account for the correlation between multiple embryos per patient. Major Findings— The average age of the patients was 35.04 yrs. The average patient age associated with euploid and aneuploid embryos was not different (P = 0.6454). We found a significant difference in c45 = t5-t4 (P = 0.0298). Our results indicated this interval on average lasts significantly longer for aneuploid embryos - c45(aneuploid) = 11.93hr vs c45(euploid) = 7.97hr. In a separate analysis limited to embryos from the same patients (patients = 47, total embryos=200, euploid=112, aneuploid=88), we obtained the same results (P = 0.0316). The statistical power for this analysis exceeded 87%. No other variable was different between the two groups. Conclusion— Our results demonstrate the importance of morphokinetic variables as potential biomarkers that could aid in non-invasively characterizing euploid and aneuploid embryos. We seek to study a larger population of embryos and incorporate the embryo quality in future studies.

Keywords: IVF, embryo, euploidy, aneuploidy, morphokinteic

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154 Defective Autophagy Disturbs Neural Migration and Network Activity in hiPSC-Derived Cockayne Syndrome B Disease Models

Authors: Julia Kapr, Andrea Rossi, Haribaskar Ramachandran, Marius Pollet, Ilka Egger, Selina Dangeleit, Katharina Koch, Jean Krutmann, Ellen Fritsche

Abstract:

It is widely acknowledged that animal models do not always represent human disease. Especially human brain development is difficult to model in animals due to a variety of structural and functional species-specificities. This causes significant discrepancies between predicted and apparent drug efficacies in clinical trials and their subsequent failure. Emerging alternatives based on 3D in vitro approaches, such as human brain spheres or organoids, may in the future reduce and ultimately replace animal models. Here, we present a human induced pluripotent stem cell (hiPSC)-based 3D neural in a vitro disease model for the Cockayne Syndrome B (CSB). CSB is a rare hereditary disease and is accompanied by severe neurologic defects, such as microcephaly, ataxia and intellectual disability, with currently no treatment options. Therefore, the aim of this study is to investigate the molecular and cellular defects found in neural hiPSC-derived CSB models. Understanding the underlying pathology of CSB enables the development of treatment options. The two CSB models used in this study comprise a patient-derived hiPSC line and its isogenic control as well as a CSB-deficient cell line based on a healthy hiPSC line (IMR90-4) background thereby excluding genetic background-related effects. Neurally induced and differentiated brain sphere cultures were characterized via RNA Sequencing, western blot (WB), immunocytochemistry (ICC) and multielectrode arrays (MEAs). CSB-deficiency leads to an altered gene expression of markers for autophagy, focal adhesion and neural network formation. Cell migration was significantly reduced and electrical activity was significantly increased in the disease cell lines. These data hint that the cellular pathologies is possibly underlying CSB. By induction of autophagy, the migration phenotype could be partially rescued, suggesting a crucial role of disturbed autophagy in defective neural migration of the disease lines. Altered autophagy may also lead to inefficient mitophagy. Accordingly, disease cell lines were shown to have a lower mitochondrial base activity and a higher susceptibility to mitochondrial stress induced by rotenone. Since mitochondria play an important role in neurotransmitter cycling, we suggest that defective mitochondria may lead to altered electrical activity in the disease cell lines. Failure to clear the defective mitochondria by mitophagy and thus missing initiation cues for new mitochondrial production could potentiate this problem. With our data, we aim at establishing a disease adverse outcome pathway (AOP), thereby adding to the in-depth understanding of this multi-faced disorder and subsequently contributing to alternative drug development.

Keywords: autophagy, disease modeling, in vitro, pluripotent stem cells

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153 Application of Harris Hawks Optimization Metaheuristic Algorithm and Random Forest Machine Learning Method for Long-Term Production Scheduling Problem under Uncertainty in Open-Pit Mines

Authors: Kamyar Tolouei, Ehsan Moosavi

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In open-pit mines, the long-term production scheduling optimization problem (LTPSOP) is a complicated problem that contains constraints, large datasets, and uncertainties. Uncertainty in the output is caused by several geological, economic, or technical factors. Due to its dimensions and NP-hard nature, it is usually difficult to find an ideal solution to the LTPSOP. The optimal schedule generally restricts the ore, metal, and waste tonnages, average grades, and cash flows of each period. Past decades have witnessed important measurements of long-term production scheduling and optimal algorithms since researchers have become highly cognizant of the issue. In fact, it is not possible to consider LTPSOP as a well-solved problem. Traditional production scheduling methods in open-pit mines apply an estimated orebody model to produce optimal schedules. The smoothing result of some geostatistical estimation procedures causes most of the mine schedules and production predictions to be unrealistic and imperfect. With the expansion of simulation procedures, the risks from grade uncertainty in ore reserves can be evaluated and organized through a set of equally probable orebody realizations. In this paper, to synthesize grade uncertainty into the strategic mine schedule, a stochastic integer programming framework is presented to LTPSOP. The objective function of the model is to maximize the net present value and minimize the risk of deviation from the production targets considering grade uncertainty simultaneously while satisfying all technical constraints and operational requirements. Instead of applying one estimated orebody model as input to optimize the production schedule, a set of equally probable orebody realizations are applied to synthesize grade uncertainty in the strategic mine schedule and to produce a more profitable and risk-based production schedule. A mixture of metaheuristic procedures and mathematical methods paves the way to achieve an appropriate solution. This paper introduced a hybrid model between the augmented Lagrangian relaxation (ALR) method and the metaheuristic algorithm, the Harris Hawks optimization (HHO), to solve the LTPSOP under grade uncertainty conditions. In this study, the HHO is experienced to update Lagrange coefficients. Besides, a machine learning method called Random Forest is applied to estimate gold grade in a mineral deposit. The Monte Carlo method is used as the simulation method with 20 realizations. The results specify that the progressive versions have been considerably developed in comparison with the traditional methods. The outcomes were also compared with the ALR-genetic algorithm and ALR-sub-gradient. To indicate the applicability of the model, a case study on an open-pit gold mining operation is implemented. The framework displays the capability to minimize risk and improvement in the expected net present value and financial profitability for LTPSOP. The framework could control geological risk more effectively than the traditional procedure considering grade uncertainty in the hybrid model framework.

Keywords: grade uncertainty, metaheuristic algorithms, open-pit mine, production scheduling optimization

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152 Networked Media, Citizen Journalism and Political Participation in Post-Revolutionary Tunisia: Insight from a European Research Project

Authors: Andrea Miconi

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The research will focus on the results of the Tempus European Project eMEDia dedicated to Cross-Media Journalism. The project is founded by the European Commission as it involves four European partners - IULM University, Tampere University, University of Barcelona, and the Mediterranean network Unimed - and three Tunisian Universities – IPSI La Manouba, Sfax and Sousse – along with the Tunisian Ministry for Higher Education and the National Syndicate of Journalists. The focus on Tunisian condition is basically due to the role played by digital activists in its recent history. The research is dedicated to the relationship between political participation, news-making practices and the spread of social media, as it is affecting Tunisian society. As we know, Tunisia during the Arab Spring had been widely considered as a laboratory for the analysis the use of new technologies for political participation. Nonetheless, the literature about the Arab Spring actually fell short in explaining the genesis of the phenomenon, on the one hand by isolating technologies as a casual factor in the spread of demonstrations, and on the other by analyzing North-African condition through a biased perspective. Nowadays, it is interesting to focus on the consolidation of the information environment three years after the uprisings. And what is relevant, only a close, in-depth analysis of Tunisian society is able to provide an explanation of its history, and namely of the part of digital media in the overall evolution of political system. That is why the research is based on different methodologies: desk stage, interviews, and in-depth analysis of communication practices. Networked journalism is the condition determined by the technological innovation on news-making activities: a condition upon which professional journalist can no longer be considered the only player in the information arena, and a new skill must be developed. Along with democratization, nonetheless, the so-called citizen journalism is also likely to produce some ambiguous effects, such as the lack of professional standards and the spread of information cascades, which may prove to be particularly dangerous in an evolving media market as the Tunisian one. This is why, according to the project, a new profile must be defined, which is able to manage this new condition, and which can be hardly reduced to the parameters of traditional journalistic work. Rather than simply using new devices for news visualization, communication professionals must also be able to dialogue with all new players and to accept the decentralized nature of digital environments. This networked nature of news-making seemed to emerge during the Tunisian revolution, when bloggers, journalists, and activists used to retweet each other. Nonetheless, this intensification of communication exchange was inspired by the political climax of the uprising, while all media, by definition, are also supposed to bring some effects on people’s state of mind, culture and daily life routines. That is why it is worth analyzing the consolidation of these practices in a normal, post-revolutionary situation.

Keywords: cross-media, education, Mediterranean, networked journalism, social media, Tunisia

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151 Evaluation of Bagh Printing Motifs and Processes of Madhya Pradesh: From Past to Contemporary

Authors: Kaveri Dutta, Ratna Sharma

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Indian traditional textile is a synthesis of various cultures. Art and crafts of a country showcases the rich cultural and artistic history of that nation. Prehistorically Indian handicrafts were basically made for day to day use; the yearning for aesthetic application soon saw the development of flooding designs and motifs. Similarly, Bagh print a traditional hand block Print with natural colours an Indian handicraft practiced in Bagh, Madhya Pradesh(India). Bagh print has its roots in Sindh, which is now a part of Pakistan. The present form of Bagh printing actually started in 1962 when the craftsmen migrated from Manavar to the neighboring town of Bagh situated in Madhya Pradesh and hence Bagh has always been associated with this printing style. Bagh printing basically involved blocks that are carved onto motifs that represent flora such as Jasmine, Mushroom leheriya and so on. There are some prints that were inspired by the jaali work that embellished the Taj Mahal and various other forts. Inspiration is also drawn from the landscapes and geometrical figures. The motifs evoke various moods in the serenity of the prints and that is the catchy element of Bagh prints. The development in this traditional textile is as essential as in another field. Nowadays fashion trends are fragile and innovative changes over existing fashion field in the short span is the demand of times. We must make efforts to preserve this cultural heritage of arts and crafts and this is done either by documenting the various ancient traditions or by making a blend of it. Since this craft is well known over the world, but the need is to document the original motif, fabric, technology and colors used in contemporary fashion. Hence keeping above points in mind this study on bagh print textiles of Madhya Pradesh work has been formulated. The information incorporated in the paper was based on secondary data taken from relevant books, journals, museum visit and articles. Besides for the demographic details and working profile of the artisans dealt with printing, an interview schedule was carried out in three regions of Madhya Pradesh. This work of art was expressed in Cotton fabric. For this study selected traditional motifs for Bang printing was used. Some of the popular traditional Bagh motifs are Jasmine, Mushroom leheriya, geometrical figures and jaali work. The Bagh printed cotton fabrics were developed into a range of men’s ethic wear in combination with embroideries from Rajasthan. Products developed were bandhgala jackets, kurtas, serwani and dupattas. From the present study, it can be observed that the embellished traditional Bang printed range of ethnic men’s wear resulted in the fresh and colourful pattern. The embroidered Bagh printed cotton fabric also created a huge change in a positive way among artisans of the three regions.

Keywords: art and craft of Madhya Pradesh, evolution of printing in India, history of Bagh printing, sources of inspiration

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150 Influence of Cryo-Grinding on Particle Size Distribution of Proso Millet Bran Fraction

Authors: Maja Benkovic, Dubravka Novotni, Bojana Voucko, Duska Curic, Damir Jezek, Nikolina Cukelj

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Cryo-grinding is an ultra-fine grinding method used in the pharmaceutical industry, production of herbs and spices and in the production and handling of cereals, due to its ability to produce powders with small particle sizes which maintain their favorable bioactive profile. The aim of this study was to determine the particle size distributions of the proso millet (Panicum miliaceum) bran fraction grinded at cryogenic temperature (using liquid nitrogen (LN₂) cooling, T = - 196 °C), in comparison to non-cooled grinding. Proso millet bran is primarily used as an animal feed, but has a potential in food applications, either as a substrate for extraction of bioactive compounds or raw material in the bakery industry. For both applications finer particle sizes of the bran could be beneficial. Thus, millet bran was ground for 2, 4, 8 and 12 minutes using the ball mill (CryoMill, Retsch GmbH, Haan, Germany) at three grinding modes: (I) without cooling, (II) at cryo-temperature, and (III) at cryo-temperature with included 1 minute of intermediate cryo-cooling step after every 2 minutes of grinding, which is usually applied when samples require longer grinding times. The sample was placed in a 50 mL stainless steel jar containing one grinding ball (Ø 25 mm). The oscillation frequency in all three modes was 30 Hz. Particle size distributions of the bran were determined by a laser diffraction particle sizing method (Mastersizer 2000) using the Scirocco 2000 dry dispersion unit (Malvern Instruments, Malvern, UK). Three main effects of the grinding set-up were visible from the results. Firstly, grinding time at all three modes had a significant effect on all particle size parameters: d(0.1), d(0.5), d(0.9), D[3,2], D[4,3], span and specific surface area. Longer grinding times resulted in lower values of the above-listed parameters, e.g. the averaged d(0.5) of the sample (229.57±1.46 µm) dropped to 51.29±1.28 µm after 2 minutes grinding without LN₂, and additionally to 43.00±1.33 µm after 4 minutes of grinding without LN₂. The only exception was the sample ground for 12 minutes without cooling, where an increase in particle diameters occurred (d(0.5)=62.85±2.20 µm), probably due to particles adhering to one another and forming larger particle clusters. Secondly, samples with LN₂ cooling exhibited lower diameters in comparison to non-cooled. For example, after 8 minutes of non-cooled grinding d(0.5)=46.97±1.05 µm was achieved, while the LN₂ cooling enabled collection of particles with average sizes of d(0.5)=18.57±0.18 µm. Thirdly, the application of intermediate cryo-cooling step resulted in similar particle diameters (d(0.5)=15.83±0.36 µm, 12 min of grinding) as cryo-milling without this step (d(0.5)=16.33±2.09 µm, 12 min of grinding). This indicates that intermediate cooling is not necessary for the current application, which consequently reduces the consumption of LN₂. These results point out the potential beneficial effects of millet bran grinding at cryo-temperatures. Further research will show if the lower particle size achieved in comparison to non-cooled grinding could result in increased bioavailability of bioactive compounds, as well as protein digestibility and solubility of dietary fibers of the proso millet bran fraction.

Keywords: ball mill, cryo-milling, particle size distribution, proso millet (Panicum miliaceum) bran

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149 Atypical Intoxication Due to Fluoxetine Abuse with Symptoms of Amnesia

Authors: Ayse Gul Bilen

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Selective serotonin reuptake inhibitors (SSRIs) are commonly prescribed antidepressants that are used clinically for the treatment of anxiety disorders, obsessive-compulsive disorder (OCD), panic disorders and eating disorders. The first SSRI, fluoxetine (sold under the brand names Prozac and Sarafem among others), had an adverse effect profile better than any other available antidepressant when it was introduced because of its selectivity for serotonin receptors. They have been considered almost free of side effects and have become widely prescribed, however questions about the safety and tolerability of SSRIs have emerged with their continued use. Most SSRI side effects are dose-related and can be attributed to serotonergic effects such as nausea. Continuous use might trigger adverse effects such as hyponatremia, tremor, nausea, weight gain, sleep disturbance and sexual dysfunction. Moderate toxicity can be safely observed in the hospital for 24 hours, and mild cases can be safely discharged (if asymptomatic) from the emergency department once cleared by Psychiatry in cases of intentional overdose and after 6 to 8 hours of observation. Although fluoxetine is relatively safe in terms of overdose, it might still be cardiotoxic and inhibit platelet secretion, aggregation, and plug formation. There have been reported clinical cases of seizures, cardiac conduction abnormalities, and even fatalities associated with fluoxetine ingestions. While the medical literature strongly suggests that most fluoxetine overdoses are benign, emergency physicians need to remain cognizant that intentional, high-dose fluoxetine ingestions may induce seizures and can even be fatal due to cardiac arrhythmia. Our case is a 35-year old female patient who was sent to ER with symptoms of confusion, amnesia and loss of orientation for time and location after being found wandering in the streets unconsciously by police forces that informed 112. Upon laboratory examination, no pathological symptom was found except sinus tachycardia in the EKG and high levels of aspartate transaminase (AST) and alanine transaminase (ALT). Diffusion MRI and computed tomography (CT) of the brain all looked normal. Upon physical and sexual examination, no signs of abuse or trauma were found. Test results for narcotics, stimulants and alcohol were negative as well. There was a presence of dysrhythmia which required admission to the intensive care unit (ICU). The patient gained back her conscience after 24 hours. It was discovered from her story afterward that she had been using fluoxetine due to post-traumatic stress disorder (PTSD) for 6 months and that she had attempted suicide after taking 3 boxes of fluoxetine due to the loss of a parent. She was then transferred to the psychiatric clinic. Our study aims to highlight the need to consider toxicologic drug use, in particular, the abuse of selective serotonin reuptake inhibitors (SSRIs), which have been widely prescribed due to presumed safety and tolerability, for diagnosis of patients applying to the emergency room (ER).

Keywords: abuse, amnesia, fluoxetine, intoxication, SSRI

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148 Quantification of the Non-Registered Electrical and Electronic Equipment for Domestic Consumption and Enhancing E-Waste Estimation: A Case Study on TVs in Vietnam

Authors: Ha Phuong Tran, Feng Wang, Jo Dewulf, Hai Trung Huynh, Thomas Schaubroeck

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The fast increase and complex components have made waste of electrical and electronic equipment (or e-waste) one of the most problematic waste streams worldwide. Precise information on its size on national, regional and global level has therefore been highlighted as prerequisite to obtain a proper management system. However, this is a very challenging task, especially in developing countries where both formal e-waste management system and necessary statistical data for e-waste estimation, i.e. data on the production, sale and trade of electrical and electronic equipment (EEE), are often lacking. Moreover, there is an inflow of non-registered electronic and electric equipment, which ‘invisibly’ enters the EEE domestic market and then is used for domestic consumption. The non-registration/invisibility and (in most of the case) illicit nature of this flow make it difficult or even impossible to be captured in any statistical system. The e-waste generated from it is thus often uncounted in current e-waste estimation based on statistical market data. Therefore, this study focuses on enhancing e-waste estimation in developing countries and proposing a calculation pathway to quantify the magnitude of the non-registered EEE inflow. An advanced Input-Out Analysis model (i.e. the Sale–Stock–Lifespan model) has been integrated in the calculation procedure. In general, Sale-Stock-Lifespan model assists to improve the quality of input data for modeling (i.e. perform data consolidation to create more accurate lifespan profile, model dynamic lifespan to take into account its changes over time), via which the quality of e-waste estimation can be improved. To demonstrate the above objectives, a case study on televisions (TVs) in Vietnam has been employed. The results show that the amount of waste TVs in Vietnam has increased four times since 2000 till now. This upward trend is expected to continue in the future. In 2035, a total of 9.51 million TVs are predicted to be discarded. Moreover, estimation of non-registered TV inflow shows that it might on average contribute about 15% to the total TVs sold on the Vietnamese market during the whole period of 2002 to 2013. To tackle potential uncertainties associated with estimation models and input data, sensitivity analysis has been applied. The results show that both estimations of waste and non-registered inflow depend on two parameters i.e. number of TVs used in household and the lifespan. Particularly, with a 1% increase in the TV in-use rate, the average market share of non-register inflow in the period 2002-2013 increases 0.95%. However, it decreases from 27% to 15% when the constant unadjusted lifespan is replaced by the dynamic adjusted lifespan. The effect of these two parameters on the amount of waste TV generation for each year is more complex and non-linear over time. To conclude, despite of remaining uncertainty, this study is the first attempt to apply the Sale-Stock-Lifespan model to improve the e-waste estimation in developing countries and to quantify the non-registered EEE inflow to domestic consumption. It therefore can be further improved in future with more knowledge and data.

Keywords: e-waste, non-registered electrical and electronic equipment, TVs, Vietnam

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147 Comparison of Nutritional Status of Asthmatic vs Non-Asthmatic Adults

Authors: Ayesha Mushtaq

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Asthma is a pulmonary disease in which blockade of the airway takes place due to inflammation as a response to certain allergens. Breathing troubles, cough, and dyspnea are one of the few symptoms. Several studies have indicated a significant effect on asthma due to changes in dietary routines. Certain food items, such as oily foods and other materials, are known to cause an increase in the symptoms of asthma. Low dietary intake of fruits and vegetables may be important in relation to asthma prevalence. The objective of this study is to assess and compare the nutritional status of asthmatic and non-asthmatic patients. The significance of this study lies in the factor that it will help nutritionists to arrange a feasible dietary routine for asthmatic patients. This research was conducted at the Pulmonology Department of the Pakistan Institute of Medical Science Islamabad. About thirty hundred thirty-four million people are affected by asthma worldwide. Pakistan is on the verge of being an uplifted urban population and asthma cases are increasingly high these days. Several studies suggest an increase in the Asthmatic patient population due to improper diet. This is a cross-sectional study aimed at assessing the nutritious standing of Asthmatic and non-asthmatic patients. This research took place at the Pakistan Institute of Medical Sciences (PIMS), Islamabad, Pakistan. The research included asthmatic and non-asthmatic patients coming to the pulmonology department clinic at the Pakistan Institute of Medical Sciences (PIMS). These patients were aged between 20-60 years. A questionnaire was developed for these patients to estimate their dietary plans in these patients. The methodology included four sections. The first section was the Socio-Demographic profile, which included age, gender, monthly income and occupation. The next section was anthropometric measurements which included the weight, height and body mass index (BMI) of an individual. The next section, section three, was about the biochemical attributes, such as for biochemical profiling, pulmonary function testing (PFT) was performed. In the next section, Dietary habits were assessed by a food frequency questionnaire (FFQ) through food habits and consumption pattern was assessed. The next section life style data, in which the person's level of physical activity, sleep and smoking habits were assessed. The next section was statistical analysis. All the data obtained from the study were statistically analyzed and assessed. Most of the asthma Patients were females, with weight more than normal or even obese. Body Mass Index (BMI) was higher in asthma Patients than those in non-Asthmatic ones. When the nutritional Values were assessed, we came to know that these patients were low on certain nutrients and their diet included more junk and oily food than healthy vegetables and fruits. Beverages intake was also included in the same assessment. It is evident from this study that nutritional status has a contributory effect on asthma. So, patients on the verge of developing asthma or those who have developed asthma should focus on their diet, maintain good eating habits and take healthy diets, including fruits and vegetables rather than oily foods. Proper sleep may also contribute to the control of asthma.

Keywords: BMI, nutrition, PAL, diet

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146 Moringa olifera Curate The Toxic Potential of CuO Nanoparticles in Oreochromis mossambicus

Authors: Farhat Jabeen, Muhammad Asad

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The study assessed the curative potential of Moringa olifera seeds against copper oxide nanoparticles induced toxicity in Oreochromis mossambicus. In order to investigate the curative potential of M. olifera seeds, firstly we examine its chemical composition, secondary metabolites, and bioactive compounds including hydroxyl-cinnamic acids, flavanols and hydroxybenzoic acids through standard methods and high performance liquid chromatography. In current study, the potential sub-lethal toxic dose of CuO-NPs (0.12 mg/l) was investigated through pilot experiment and three non-lethal doses (low=32, medium=48 and high=96 mg/l) of M. olifera were selected on the basis of its LC50 value for O. mossambicus. The experimental fish, O. mossambicus (n=100 of approximately 20 g each) were procured from Manawan Fisheries Complex, Lahore, and acclimatized for two weeks in glass aquaria. Experiment was conducted in accordance with the guidelines of Institutional Animal Ethics Committee, Government College University Faisalabad, Pakistan. During acclimatization and experimental period, fish received the commercial fish feed at 2.5% body weight daily. In order to assess the curative effect of M. olifera against CuO NPs induced toxicity, O. mossambicus were randomly divided into five groups and were designated as control (C) without any treatment, positive control (G*) exposed to potential toxic dose of CuO-NPs at 0.12 mg/l, and three treated groups namely G1, G2, and G3 co-treated with 0.12 mg/l of CuO-NPs plus different doses of M. olifera seed extract at 32, 48, and 96 mg/l, respectively for 56 days. Fish were exposed to waterborne CuO NPs and M. olifera seed extract. CuO-NPs treatment was ceased after 28 days but the doses of M. olifera were continued for 56 days. Blood was taken after 28 and 56 days through caudal venipuncture. Liver and intestine were taken for oxidative stress and histological studies after 56 days. In M. olifera seeds, moisture contents, crude protein, lipids, carbohydrates and ash were recorded as 3.8, 37.83, 32.52, 46.12, and 7.75%, respectively on dry weight basis. Total energy was recorded as 627.36 kcal/100g. Qualitative analysis of M. olifera seeds showed the presence of terpenoids, saponins, flavonoids, alkaloids and phenolics, while its quantitative analysis showed the considerable amount of total phenolics, flavonoids, saponins, and alkaloids as 134.75, 170.15, 1.57, and 0.4 µg/mg, respectively. Analysis of bioactive compounds in M. olifera seeds showed the presence of hydroxy-cinnamic acids (6.07 µg/ml), flavanols (71.72 µg/ml), and hydroxyl benzoic acids (97.82 µg/ml). The results showed that M. oliefera seed extract at 48 and 56 mg/l was able to cure against the toxic effects of CuO-NPs. The significant changes were observed in G* and G1 for sero-hepatic enzymes, anti-oxidants and histological profile. The investigations of this study showed that M. olifera is a good curative agent against potential induced toxicity of CuO-NPs in O. mossambicus. The curative effect of M. olifera is attributed to the presence of higher amount of secondary metabolites and bioactive compounds. This study suggested the use of M. olifera to curate different ailments in fish and other organisms.

Keywords: CuO nanoparticles, curative, Moringa olifera, Oreochromis mossambicus

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145 Optimization of Multi-Disciplinary Expertise and Resource for End-Stage Renal Failure (ESRF) Patient Care

Authors: Mohamed Naser Zainol, P. P. Angeline Song

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Over the years, the profile of end-stage renal patients placed under The National Kidney Foundation Singapore (NKFS) dialysis program has evolved, with a gradual incline in the number of patients with behavior-related issues. With these challenging profiles, social workers and counsellors are often expected to oversee behavior management, through referrals from its partnering colleagues. Due to the segregation of tasks usually found in many hospital-based multi-disciplinary settings, social workers’ and counsellors’ interventions are often seen as an endpoint, limiting other stakeholders’ involvement that could otherwise be potentially crucial in managing such patients. While patients’ contact in local hospitals often leads to eventual discharge, NKFS patients are mostly long term. It is interesting to note that these patients are regularly seen by a team of professionals that includes doctors, nurses, dietitians, exercise specialists in NKFS. The dynamism of relationships presents an opportunity for any of these professionals to take ownership of their potentials in leading interventions that can be helpful to patients. As such, it is important to have a framework that incorporates the strength of these professionals and also channels empowerment across the multi-disciplinary team in working towards wholistic patient care. This paper would like to suggest a new framework for NKFS’s multi-disciplinary team, where the group synergy and dynamics are used to encourage ownership and promote empowerment. The social worker and counsellor use group work skills and his/her knowledge of its members’ strengths, to generate constructive solutions that are centered towards patient’s growth. Using key ideas from Karl’s Tomm Interpersonal Communications, the Communication Management of Meaning and Motivational Interviewing, the social worker and counsellor through a series of guided meeting with other colleagues, facilitates the transmission of understanding, responsibility sharing and tapping on team resources for patient care. As a result, the patient can experience personal and concerted approach and begins to flow in a direction that is helpful for him. Using seven case studies of identified patients with behavioral issues, the social worker and counsellor apply this framework for a period of six months. Patient’s overall improvement through interventions as a result of this framework are recorded using the AB single case design, with baseline measured three months before referral. Interviews with patients and their families, as well as other colleagues that are not part of the multi-disciplinary team are solicited at mid and end points to gather their experiences about patient’s progress as a by-product of this framework. Expert interviews will be conducted on each member of the multi-disciplinary team to study their observations and experience in using this new framework. Hence, this exploratory framework hopes to identify the inherent usefulness in managing patients with behavior related issues. Moreover, it would provide indicators in improving aspects of the framework when applied to a larger population.

Keywords: behavior management, end-stage renal failure, satellite dialysis, multi-disciplinary team

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144 Review of the Nutritional Value of Spirulina as a Potential Replacement of Fishmeal in Aquafeed

Authors: Onada Olawale Ahmed

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As the intensification of aquaculture production increases on global scale, the growing concern of fish farmers around the world is related to cost of fish production, where cost of feeding takes substantial percentage. Fishmeal (FM) is one of the most expensive ingredients, and its high dependence in aqua-feed production translates to high cost of feeding of stocked fish. However, to reach a sustainable aquaculture, new alternative protein sources including cheaper plant or animal origin proteins are needed to be introduced for stable aqua-feed production. Spirulina is a cyanobacterium that has good nutrient profile that could be useful in aquaculture. This review therefore emphasizes on the nutritional value of Spirulina as a potential replacement of FM in aqua-feed. Spirulina is a planktonic photosynthetic filamentous cyanobacterium that forms massive populations in tropical and subtropical bodies of water with high levels of carbonate and bicarbonate. Spirulina grows naturally in nutrient rich alkaline lake with water salinity ( > 30 g/l) and high pH (8.5–11.0). Its artificial production requires luminosity (photo-period 12/12, 4 luxes), temperature (30 °C), inoculum, water stirring device, dissolved solids (10–60 g/litre), pH (8.5– 10.5), good water quality, and macro and micronutrient presence (C, N, P, K, S, Mg, Na, Cl, Ca and Fe, Zn, Cu, Ni, Co, Se). Spirulina has also been reported to grow on agro-industrial waste such as sugar mill waste effluent, poultry industry waste, fertilizer factory waste, and urban waste and organic matter. Chemical composition of Spirulina indicates that it has high nutritional value due to its content of 55-70% protein, 14-19% soluble carbohydrate, high amount of polyunsaturated fatty acids (PUFAs), 1.5–2.0 percent of 5–6 percent total lipid, all the essential minerals are available in spirulina which contributes about 7 percent (average range 2.76–3.00 percent of total weight) under laboratory conditions, β-carotene, B-group vitamin, vitamin E, iron, potassium and chlorophyll are also available in spirulina. Spirulina protein has a balanced composition of amino acids with concentration of methionine, tryptophan and other amino acids almost similar to those of casein, although, this depends upon the culture media used. Positive effects of spirulina on growth, feed utilization and stress and disease resistance of cultured fish have been reported in earlier studies. Spirulina was reported to replace up to 40% of fishmeal protein in tilapia (Oreochromis mossambicus) diet and even higher replacement of fishmeal was possible in common carp (Cyprinus carpio), partial replacement of fish meal with spirulina in diets for parrot fish (Oplegnathus fasciatus) and Tilapia (Orechromis niloticus) has also been conducted. Spirulina have considerable potential for development, especially as a small-scale crop for nutritional enhancement and health improvement of fish. It is important therefore that more research needs to be conducted on its production, inclusion level in aqua-feed and its possible potential use of aquaculture.

Keywords: aquaculture, spirulina, fish nutrition, fish feed

Procedia PDF Downloads 490
143 Safety Considerations of Furanics for Sustainable Applications in Advanced Biorefineries

Authors: Anitha Muralidhara, Victor Engelen, Christophe Len, Pascal Pandard, Guy Marlair

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Production of bio-based chemicals and materials from lignocellulosic biomass is gaining tremendous importance in advanced bio-refineries while aiming towards progressive replacement of petroleum based chemicals in transportation fuels and commodity polymers. One such attempt has resulted in the production of key furan derivatives (FD) such as furfural, HMF, MMF etc., via acid catalyzed dehydration (ACD) of C6 and C5 sugars, which are further converted into key chemicals or intermediates (such as Furandicarboxylic acid, Furfuryl alcohol etc.,). In subsequent processes, many high potential FD are produced, that can be converted into high added value polymers or high energy density biofuels. During ACD, an unavoidable polyfuranic byproduct is generated which is called humins. The family of FD is very large with varying chemical structures and diverse physicochemical properties. Accordingly, the associated risk profiles may largely vary. Hazardous Material (Haz-mat) classification systems such as GHS (CLP in the EU) and the UN TDG Model Regulations for transport of dangerous goods are one of the preliminary requirements for all chemicals for their appropriate classification, labelling, packaging, safe storage, and transportation. Considering the growing application routes of FD, it becomes important to notice the limited access to safety related information (safety data sheets available only for famous compounds such as HMF, furfural etc.,) in these internationally recognized haz-mat classification systems. However, these classifications do not necessarily provide information about the extent of risk involved when the chemical is used in any specific application. Factors such as thermal stability, speed of combustion, chemical incompatibilities, etc., can equally influence the safety profile of a compound, that are clearly out of the scope of any haz-mat classification system. Irrespective of the bio-based origin, FD has so far received inconsistent remarks concerning their toxicity profiles. With such inconsistencies, there is a fear that, a large family of FD may also follow extreme judgmental scenarios like ionic liquids, by ranking some compounds as extremely thermally stable, non-flammable, etc., Unless clarified, these messages could lead to misleading judgements while ranking the chemical based on its hazard rating. Safety is a key aspect in any sustainable biorefinery operation/facility, which is often underscored or neglected. To fill up these existing data gaps and to address ambiguities and discrepancies, the current study focuses on giving preliminary insights on safety assessment of FD and their potential targeted by-products. With the available information in the literature and obtained experimental results, physicochemical safety, environmental safety as well as (a scenario based) fire safety profiles of key FD, as well as side streams such as humins and levulinic acid, will be considered. With this, the study focuses on defining patterns and trends that gives coherent safety related information for existing and newly synthesized FD in the market for better functionality and sustainable applications.

Keywords: furanics, humins, safety, thermal and fire hazard, toxicity

Procedia PDF Downloads 149
142 Temperature-Dependent Post-Mortem Changes in Human Cardiac Troponin-T (cTnT): An Approach in Determining Postmortem Interval

Authors: Sachil Kumar, Anoop Kumar Verma, Wahid Ali, Uma Shankar Singh

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Globally approximately 55.3 million people die each year. In the India there were 95 lakh annual deaths in 2013. The number of deaths resulted from homicides, suicides and unintentional injuries in the same period was about 5.7 lakh. The ever-increasing crime rate necessitated the development of methods for determining time since death. An erroneous time of death window can lead investigators down the wrong path or possibly focus a case on an innocent suspect. In this regard a research was carried out by analyzing the temperature dependent degradation of a Cardiac Troponin-T protein (cTnT) in the myocardium postmortem as a marker for time since death. Cardiac tissue samples were collected from (n=6) medico-legal autopsies, (in the Department of Forensic Medicine and Toxicology, King George’s Medical University, Lucknow India) after informed consent from the relatives and studied post-mortem degradation by incubation of the cardiac tissue at room temperature (20±2 OC), 12 0C, 25 0C and 37 0C for different time periods ((~5, 26, 50, 84, 132, 157, 180, 205, and 230 hours). The cases included were the subjects of road traffic accidents (RTA) without any prior history of disease who died in the hospital and their exact time of death was known. The analysis involved extraction of the protein, separation by denaturing gel electrophoresis (SDS-PAGE) and visualization by Western blot using cTnT specific monoclonal antibodies. The area of the bands within a lane was quantified by scanning and digitizing the image using Gel Doc. The data shows a distinct temporal profile corresponding to the degradation of cTnT by proteases found in cardiac muscle. The disappearance of intact cTnT and the appearance of lower molecular weight bands are easily observed. Western blot data clearly showed the intact protein at 42 kDa, two major (27 kDa, 10kDa) fragments, two additional minor fragments (32 kDa) and formation of low molecular weight fragments as time increases. At 12 0C the intensity of band (intact cTnT) decreased steadily as compared to RT, 25 0C and 37 0C. Overall, both PMI and temperature had a statistically significant effect where the greatest amount of protein breakdown was observed within the first 38 h and at the highest temperature, 37 0C. The combination of high temperature (37 0C) and long Postmortem interval (105.15 hrs) had the most drastic effect on the breakdown of cTnT. If the percent intact cTnT is calculated from the total area integrated within a Western blot lane, then the percent intact cTnT shows a pseudo-first order relationship when plotted against the log of the time postmortem. These plots show a good coefficient of correlation of r = 0.95 (p=0.003) for the regression of the human heart at different temperature conditions. The data presented demonstrates that this technique can provide an extended time range during which Postmortem interval can be more accurately estimated.

Keywords: degradation, postmortem interval, proteolysis, temperature, troponin

Procedia PDF Downloads 362
141 Cryotopic Macroporous Polymeric Matrices for Regenerative Medicine and Tissue Engineering Applications

Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

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Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

Procedia PDF Downloads 197
140 A Study of Interleukin-1β Genetic Polymorphisms in Gastric Carcinoma and Colorectal Carcinoma in Egyptian Patients

Authors: Mariam Khaled, Noha Farag, Ghada Mohamed Abdel Salam, Khaled Abu-Aisha, Mohamed El-Azizi

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Gastric and colorectal cancers are among the most frequent causes of cancer-associated mortalities in Africa. They have been considered as a global public health concern, as nearly one million new cases are reported per year. IL-1β is a pro-inflammatory cytokine-produced by activated macrophages and monocytes- and a member of the IL-1 family. The inactive IL-1β precursor is cleaved and activated by caspase-1 enzyme, which itself is activated by the assembly of intracellular structures defined as NLRP3 (Nod Like receptor P3) inflammasomes. Activated IL-1β stimulates the Interleukin-1 receptor type-1 (IL-1R1), which is responsible for the initiation of a signal transduction pathway leading to cell proliferation. It has been proven that the IL-1β gene is a highly polymorphic gene in which single nucleotide polymorphisms (SNPs) may affect its expression. It has been previously reported that SNPs including base transitions between C and T at positions, -511 (C-T; dbSNP: rs16944) and -31 (C-T; dbSNP: rs1143627), from the transcriptional start site, contribute to the pathogenesis of gastric and colorectal cancers by affecting IL-1β levels. Altered production of IL-1β due to such polymorphisms is suspected to stimulate an amplified inflammatory response and promote Epithelial Mesenchymal Transition leading to malignancy. Allele frequency distribution of the IL-1β-31 and -511 SNPs, in different populations, and their correlation to the incidence of gastric and colorectal cancers, has been intriguing to researchers worldwide. The current study aims to investigate allele distributions of the IL-1β SNPs among gastric and colorectal cancers Egyptian patients. In order to achieve to that, 89 Biopsy and surgical specimens from the antrum and corpus mucosa of chronic gastritis subjects and gastric and colorectal carcinoma patients was collected for DNA extraction followed by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). The amplified PCR products of IL-1β-31C > T and IL-1β-511T > C were digested by incubation with the restriction endonuclease enzymes ALu1 and Ava1. Statistical analysis was carried out to determine the allele frequency distribution in the three studied groups. Also, the effect of the IL-1β -31 and -511 SNPs on nuclear factor binding was analyzed using Fluorescence Electrophoretic Mobility Shift Assay (EMSA), preceded by nuclear factor extraction from gastric and colorectal tissue samples and LPS stimulated monocytes. The results of this study showed that a significantly higher percentage of Egyptian gastric cancer patients have a homozygous CC genotype at the IL-1β-31 position and a heterozygous TC genotype at the IL-1β-511 position. Moreover, a significantly higher percentage of the colorectal cancer patients have a homozygous CC genotype at the IL-1β-31 and -511 positions as compared to the control group. In addition, the EMSA results showed that IL-1β-31C/T and IL-1β-511T/C SNPs do not affect nuclear factor binding. Results of this study suggest that the IL-1β-31 C/T and IL-1β-511 T/C may be correlated to the incidence of gastric cancer in Egyptian patients; however, similar findings couldn’t be proven in the colorectal cancer patients group for the IL-1β-511 T/C SNP. This is the first study to investigate IL-1β -31 and -511 SNPs in the Egyptian population.

Keywords: colorectal cancer, Egyptian patients, gastric cancer, interleukin-1β, single nucleotide polymorphisms

Procedia PDF Downloads 113
139 School Accidents in Educational Establishment in Tunisia: A Five Years Retrospective Survey in the Governorate of Mahdia

Authors: Lamia Bouzgarrou, Amira Omrane, Leila Mrabet, Taoufik Khalfallah

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Background and aims: School accidents are one of the leading causes of morbidity and mortality among pupils and students. Indeed, they may induce an elevated number of lost school days, heavy emotional and physical disabilities, and financial costs on the victims and their families. This study aims to evaluate the annual incidence of school accidents in the central Tunisian governorate of Mahdia and to identify the epidemiological profile of victims and risk factors of these accidents. Methods: A retrospective study was conducted over the period of 5 school years, focusing on school accidents that occurred in public educational institutions (primary, basic, secondary and university) in the governorate of Mahdia (area = 2 966 km² and number of inhabitants in 2014 = 410 812). All accidents declared near the only official insurance of this type of injuries (MASU: Mutual School and University Accidents), and initially taken in charge at the University Hospital of Mahdia were included. Data was collected from the MASU reporting forms and the medical records of emergency and other specialized hospital departments. Results: With 3248 identified victims, the annual incidence of school accidents was equal to 0.69 per 100 pupils and students per year. The average age of victims was 14.51 ± 0.059 years and the sex ratio was 1.58. Pupils aged between 12 and 15 years, were concerned by 46.7% of the identified accidents. The practice of sports was the most relevant circumstances of these accidents (76.2 %). In 56.58 % of cases, falls were the leading mechanism. Bruises and fractures were the most frequent lesions (32.43 % and 30.51 %). Serious school accidents were noted in 28% of cases with hospitalization in 2.27 % of them. The average lost school days, was 12.23±1.73 days. Accidents occurring during sports or leisure activities were significantly more serious (p= 0.021). Furthermore, the frequency of hospitalization was significantly higher among boys (2.81% vs. 1.43%; p= 0.035), students ≤11 years (p= 0.008), and following crush trauma (p= 0.000). In addition, the surgical interventions were statistically more frequent among male victims (p=0.00), accidents occurring during physical education sessions (p=0.000); those associated to falls (p=0.000) and to crushes mechanisms (p=0.002), and injuries affecting lower limbs (p=0.000). Following this Multi-varied analysis concluded that the severity of school accident is correlated to the activity practiced during the trauma and the geographical location of the school. Conclusion: Children and adolescents are one of the most vulnerable groups against incidents with the risk of permanent disability, mainly related to the perturbation of the growth process and physiological limitations. Our five-year study, objectified a real elevate incidence of school accident among children and adolescents, with a considerable rate of severe injuries. In any community, the promotion of adolescents and children’s health is an important indicator of the public health level. Thus, it’s important to develop a multidisciplinary prevention strategy of school accident, based on safety and security rules and adapted to the specificity of our context.

Keywords: children and adolescents, children health, injuries and disability, school accident

Procedia PDF Downloads 96
138 A Novel Upregulated circ_0032746 on Sponging with MIR4270 Promotes the Proliferation and Migration of Esophageal Squamous Cell Carcinoma

Authors: Sachin Mulmi Shrestha, Xin Fang, Hui Ye, Lihua Ren, Qinghua Ji, Ruihua Shi

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Background: Esophageal squamous cell carcinoma (ESCC) is a tumor arising from esophageal epithelial cells and is one of the major disease subtype in Asian countries, including China. Esophageal cancer is the 7th highest incidence based on the 2020 data of GLOBOCAN. The pathogenesis of cancer is still not well understood as many molecular and genetic basis of esophageal carcinogenesis has yet to be clearly elucidated. Circular RNAs are RNA molecules that are formed by back-splicing covalently joined 3′- and 5′-endsrather than canonical splicing, and recent data suggest circular RNAs could sponge miRNAs and are enriched with functional miRNA binding sites. Hence, we studied the mechanism of circular RNA, its biological function, and the relationship between microRNA in the carcinogenesis of ESCC. Methods: 4 pairs of normal and esophageal cancer tissues were collected in Zhongda hospital, affiliated to Southeast University, and high-throughput RNA sequencing was done. The result revealed that circ_0032746 was upregulated, and thus we selected circ_0032746 for further study. The backsplice junction of circRNA was validated by sanger sequence, and stability was determined by RNASE R assay. The binding site of circRNA and microRNA was predicted by circinteractome,mirandaand RNAhybrid database. Furthermore, circRNA was silenced by siRNA and then by lentivirus. The regulatory axis of circ0032746/miR4270 was validated by shRNA, mimic, and inhibitor transfection. Then, in vitro experiments were performed to assess the role of circ0032746 on proliferation (CCK-8 assay and colon formation assay), migration and invasion (Transewell assay), and apoptosis of ESCC. Results: The upregulated circ0032746 was validated in 9 pairs of tissues and 5 types of cell lines by qPCR, which showed high expression and was statistically significant (P<0.005) ). Upregulated circ0032746 was silenced by shRNA, which showed significant knockdown in KYSE 30 and TE-1 cell lines expression compared to control. Nuclear and cytoplasmic mRNA fraction experiment displayed the cytoplasmic location of circ0032746. The sponging of miR4270 was validated by co-transfection of sh-circ0032746 and mimic or inhibitor. Transfection with mimic showed the decreased expression of circ_0032746, whereas inhibitor inhibited the result. In vitro experiments showed that silencing of circ_0032746 inhibited the proliferation, migration, and invasion compared to the negative control group. The apoptosis was seen higher in a knockdown group than in the control group. Furthermore, 11 common mircoRNA target mRNAs were predicted by Targetscan, MirTarbase, and miRanda database, which may further play role in the pathogenesis. Conclusion: Our results showed that novel circ_0032746 is upregulated in ESCC and plays role in itsoncogenicity. Silencing of circ_0032746 inhibits the proliferation and migration of ESCC whereas increases the apoptosis of cancer cells. Hence, circ0032746 acts as an oncogene in ESCC by sponging with miR4270 and could be a potential biomarker in the diagnosis of ESCC in the future.

Keywords: circRNA, esophageal squamous cell carcinoma, microRNA, upregulated

Procedia PDF Downloads 85
137 Assessing Brain Targeting Efficiency of Ionisable Lipid Nanoparticles Encapsulating Cas9 mRNA/gGFP Following Different Routes of Administration in Mice

Authors: Meiling Yu, Nadia Rouatbi, Khuloud T. Al-Jamal

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Background: Treatment of neurological disorders with modern medical and surgical approaches remains difficult. Gene therapy, allowing the delivery of genetic materials that encodes potential therapeutic molecules, represents an attractive option. The treatment of brain diseases with gene therapy requires the gene-editing tool to be delivered efficiently to the central nervous system. In this study, we explored the efficiency of different delivery routes, namely intravenous (i.v.), intra-cranial (i.c.), and intra-nasal (i.n.), to deliver stable nucleic acid-lipid particles (SNALPs) containing gene-editing tools namely Cas9 mRNA and sgRNA encoding for GFP as a reporter protein. We hypothesise that SNALPs can reach the brain and perform gene-editing to different extents depending on the administration route. Intranasal administration (i.n.) offers an attractive and non-invasive way to access the brain circumventing the blood–brain barrier. Successful delivery of gene-editing tools to the brain offers a great opportunity for therapeutic target validation and nucleic acids therapeutics delivery to improve treatment options for a range of neurodegenerative diseases. In this study, we utilised Rosa26-Cas9 knock-in mice, expressing GFP, to study brain distribution and gene-editing efficiency of SNALPs after i.v.; i.c. and i.n. routes of administration. Methods: Single guide RNA (sgRNA) against GFP has been designed and validated by in vitro nuclease assay. SNALPs were formulated and characterised using dynamic light scattering. The encapsulation efficiency of nucleic acids (NA) was measured by RiboGreen™ assay. SNALPs were incubated in serum to assess their ability to protect NA from degradation. Rosa26-Cas9 knock-in mice were i.v., i.n., or i.c. administered with SNALPs to test in vivo gene-editing (GFP knockout) efficiency. SNALPs were given as three doses of 0.64 mg/kg sgGFP following i.v. and i.n. or a single dose of 0.25 mg/kg sgGFP following i.c.. knockout efficiency was assessed after seven days using Sanger Sequencing and Inference of CRISPR Edits (ICE) analysis. In vivo, the biodistribution of DiR labelled SNALPs (SNALPs-DiR) was assessed at 24h post-administration using IVIS Lumina Series III. Results: Serum-stable SNALPs produced were 130-140 nm in diameter with ~90% nucleic acid loading efficiency. SNALPs could reach and stay in the brain for up to 24h following i.v.; i.n. and i.c. administration. Decreasing GFP expression (around 50% after i.v. and i.c. and 20% following i.n.) was confirmed by optical imaging. Despite the small number of mice used, ICE analysis confirmed GFP knockout in mice brains. Additional studies are currently taking place to increase mice numbers. Conclusion: Results confirmed efficient gene knockout achieved by SNALPs in Rosa26-Cas9 knock-in mice expressing GFP following different routes of administrations in the following order i.v.= i.c.> i.n. Each of the administration routes has its pros and cons. The next stages of the project involve assessing gene-editing efficiency in wild-type mice and replacing GFP as a model target with therapeutic target genes implicated in Motor Neuron Disease pathology.

Keywords: CRISPR, nanoparticles, brain diseases, administration routes

Procedia PDF Downloads 67
136 Solar Power Forecasting for the Bidding Zones of the Italian Electricity Market with an Analog Ensemble Approach

Authors: Elena Collino, Dario A. Ronzio, Goffredo Decimi, Maurizio Riva

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The rapid increase of renewable energy in Italy is led by wind and solar installations. The 2017 Italian energy strategy foresees a further development of these sustainable technologies, especially solar. This fact has resulted in new opportunities, challenges, and different problems to deal with. The growth of renewables allows to meet the European requirements regarding energy and environmental policy, but these types of sources are difficult to manage because they are intermittent and non-programmable. Operationally, these characteristics can lead to instability on the voltage profile and increasing uncertainty on energy reserve scheduling. The increasing renewable production must be considered with more and more attention especially by the Transmission System Operator (TSO). The TSO, in fact, every day provides orders on energy dispatch, once the market outcome has been determined, on extended areas, defined mainly on the basis of power transmission limitations. In Italy, six market zone are defined: Northern-Italy, Central-Northern Italy, Central-Southern Italy, Southern Italy, Sardinia, and Sicily. An accurate hourly renewable power forecasting for the day-ahead on these extended areas brings an improvement both in terms of dispatching and reserve management. In this study, an operational forecasting tool of the hourly solar output for the six Italian market zones is presented, and the performance is analysed. The implementation is carried out by means of a numerical weather prediction model, coupled with a statistical post-processing in order to derive the power forecast on the basis of the meteorological projection. The weather forecast is obtained from the limited area model RAMS on the Italian territory, initialized with IFS-ECMWF boundary conditions. The post-processing calculates the solar power production with the Analog Ensemble technique (AN). This statistical approach forecasts the production using a probability distribution of the measured production registered in the past when the weather scenario looked very similar to the forecasted one. The similarity is evaluated for the components of the solar radiation: global (GHI), diffuse (DIF) and direct normal (DNI) irradiation, together with the corresponding azimuth and zenith solar angles. These are, in fact, the main factors that affect the solar production. Considering that the AN performance is strictly related to the length and quality of the historical data a training period of more than one year has been used. The training set is made by historical Numerical Weather Prediction (NWP) forecasts at 12 UTC for the GHI, DIF and DNI variables over the Italian territory together with corresponding hourly measured production for each of the six zones. The AN technique makes it possible to estimate the aggregate solar production in the area, without information about the technologic characteristics of the all solar parks present in each area. Besides, this information is often only partially available. Every day, the hourly solar power forecast for the six Italian market zones is made publicly available through a website.

Keywords: analog ensemble, electricity market, PV forecast, solar energy

Procedia PDF Downloads 127
135 Genomic and Proteomic Variability in Glycine Max Genotypes in Response to Salt Stress

Authors: Faheema Khan

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To investigate the ability of sensitive and tolerant genotype of Glycine max to adapt to a saline environment in a field, we examined the growth performance, water relation and activities of antioxidant enzymes in relation to photosynthetic rate, chlorophyll a fluorescence, photosynthetic pigment concentration, protein and proline in plants exposed to salt stress. Ten soybean genotypes (Pusa-20, Pusa-40, Pusa-37, Pusa-16, Pusa-24, Pusa-22, BRAGG, PK-416, PK-1042, and DS-9712) were selected and grown hydroponically. After 3 days of proper germination, the seedlings were transferred to Hoagland’s solution (Hoagland and Arnon 1950). The growth chamber was maintained at a photosynthetic photon flux density of 430 μmol m−2 s−1, 14 h of light, 10 h of dark and a relative humidity of 60%. The nutrient solution was bubbled with sterile air and changed on alternate days. Ten-day-old seedlings were given seven levels of salt in the form of NaCl viz., T1 = 0 mM NaCl, T2=25 mM NaCl, T3=50 mM NaCl, T4=75 mM NaCl, T5=100 mM NaCl, T6=125 mM NaCl, T7=150 mM NaCl. The investigation showed that genotype Pusa-24, PK-416 and Pusa-20 appeared to be the most salt-sensitive. genotypes as inferred from their significantly reduced length, fresh weight and dry weight in response to the NaCl exposure. Pusa-37 appeared to be the most tolerant genotype since no significant effect of NaCl treatment on growth was found. We observed a greater decline in the photosynthetic variables like photosynthetic rate, chlorophyll fluorescence and chlorophyll content, in salt-sensitive (Pusa-24) genotype than in salt-tolerant Pusa-37 under high salinity. Numerous primers were verified on ten soybean genotypes obtained from Operon technologies among which 30 RAPD primers shown high polymorphism and genetic variation. The Jaccard’s similarity coefficient values for each pairwise comparison between cultivars were calculated and similarity coefficient matrix was constructed. The closer varieties in the cluster behaved similar in their response to salinity tolerance. Intra-clustering within the two clusters precisely grouped the 10 genotypes in sub-cluster as expected from their physiological findings.Salt tolerant genotype Pusa-37, was further analysed by 2-Dimensional gel electrophoresis to analyse the differential expression of proteins at high salt stress. In the Present study, 173 protein spots were identified. Of these, 40 proteins responsive to salinity were either up- or down-regulated in Pusa-37. Proteomic analysis in salt-tolerant genotype (Pusa-37) led to the detection of proteins involved in a variety of biological processes, such as protein synthesis (12 %), redox regulation (19 %), primary and secondary metabolism (25 %), or disease- and defence-related processes (32 %). In conclusion, the soybean plants in our study responded to salt stress by changing their protein expression pattern. The photosynthetic, biochemical and molecular study showed that there is variability in salt tolerance behaviour in soybean genotypes. Pusa-24 is the salt-sensitive and Pusa-37 is the salt-tolerant genotype. Moreover this study gives new insights into the salt-stress response in soybean and demonstrates the power of genomic and proteomic approach in plant biology studies which finally could help us in identifying the possible regulatory switches (gene/s) controlling the salt tolerant genotype of the crop plants and their possible role in defence mechanism.

Keywords: glycine max, salt stress, RAPD, genomic and proteomic variability

Procedia PDF Downloads 395
134 Absolute Quantification of the Bexsero Vaccine Component Factor H Binding Protein (fHbp) by Selected Reaction Monitoring: The Contribution of Mass Spectrometry in Vaccinology

Authors: Massimiliano Biagini, Marco Spinsanti, Gabriella De Angelis, Sara Tomei, Ilaria Ferlenghi, Maria Scarselli, Alessia Biolchi, Alessandro Muzzi, Brunella Brunelli, Silvana Savino, Marzia M. Giuliani, Isabel Delany, Paolo Costantino, Rino Rappuoli, Vega Masignani, Nathalie Norais

Abstract:

The gram-negative bacterium Neisseria meningitidis serogroup B (MenB) is an exclusively human pathogen representing the major cause of meningitides and severe sepsis in infants and children but also in young adults. This pathogen is usually present in the 30% of healthy population that act as a reservoir, spreading it through saliva and respiratory fluids during coughing, sneezing, kissing. Among surface-exposed protein components of this diplococcus, factor H binding protein is a lipoprotein proved to be a protective antigen used as a component of the recently licensed Bexsero vaccine. fHbp is a highly variable meningococcal protein: to reflect its remarkable sequence variability, it has been classified in three variants (or two subfamilies), and with poor cross-protection among the different variants. Furthermore, the level of fHbp expression varies significantly among strains, and this has also been considered an important factor for predicting MenB strain susceptibility to anti-fHbp antisera. Different methods have been used to assess fHbp expression on meningococcal strains, however, all these methods use anti-fHbp antibodies, and for this reason, the results are affected by the different affinity that antibodies can have to different antigenic variants. To overcome the limitations of an antibody-based quantification, we developed a quantitative Mass Spectrometry (MS) approach. Selected Reaction Monitoring (SRM) recently emerged as a powerful MS tool for detecting and quantifying proteins in complex mixtures. SRM is based on the targeted detection of ProteoTypicPeptides (PTPs), which are unique signatures of a protein that can be easily detected and quantified by MS. This approach, proven to be highly sensitive, quantitatively accurate and highly reproducible, was used to quantify the absolute amount of fHbp antigen in total extracts derived from 105 clinical isolates, evenly distributed among the three main variant groups and selected to be representative of the fHbp circulating subvariants around the world. We extended the study at the genetic level investigating the correlation between the differential level of expression and polymorphisms present within the genes and their promoter sequences. The implications of fHbp expression on the susceptibility of the strain to killing by anti-fHbp antisera are also presented. To date this is the first comprehensive fHbp expression profiling in a large panel of Neisseria meningitidis clinical isolates driven by an antibody-independent MS-based methodology, opening the door to new applications in vaccine coverage prediction and reinforcing the molecular understanding of released vaccines.

Keywords: quantitative mass spectrometry, Neisseria meningitidis, vaccines, bexsero, molecular epidemiology

Procedia PDF Downloads 282
133 Autophagy Promotes Vascular Smooth Muscle Cell Migration in vitro and in vivo

Authors: Changhan Ouyang, Zhonglin Xie

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In response to proatherosclerotic factors such as oxidized lipids, or to therapeutic interventions such as angioplasty, stents, or bypass surgery, vascular smooth muscle cells (VSMCs) migrate from the media to the intima, resulting in intimal hyperplasia, restenosis, graft failure, or atherosclerosis. These proatherosclerotic factors also activate autophagy in VSMCs. However, the functional role of autophagy in vascular health and disease remains poorly understood. In the present study, we determined the role of autophagy in the regulation of VSMC migration. Autophagy activity in cultured human aortic smooth muscle cells (HASMCs) and mouse carotid arteries was measured by Western blot analysis of microtubule-associated protein 1 light chain 3 B (LC3B) and P62. The VSMC migration was determined by scratch wound assay and transwell migration assay. Ex vivo smooth muscle cell migration was determined using aortic ring assay. The in vivo SMC migration was examined by staining the carotid artery sections with smooth muscle alpha actin (alpha SMA) after carotid artery ligation. To examine the relationship between autophagy and neointimal hyperplasia, C57BL/6J mice were subjected to carotid artery ligation. Seven days after injury, protein levels of Atg5, Atg7, Beclin1, and LC3B drastically increased and remained higher in the injured arteries three weeks after the injury. In parallel with the activation of autophagy, vascular injury-induced neointimal hyperplasia as estimated by increased intima/media ratio. The en face staining of carotid artery showed that vascular injury enhanced alpha SMA staining in the intimal cells as compared with the sham operation. Treatment of HASMCs with platelet-derived growth factor (PDGF), one of the major factors for vascular remodeling in response to vascular injury, increased Atg7 and LC3 II protein levels and enhanced autophagosome formation. In addition, aortic ring assay demonstrated that PDGF treated aortic rings displayed an increase in neovessel formation compared with control rings. Whole mount staining for CD31 and alpha SMA in PDGF treated neovessels revealed that the neovessel structures were stained by alpha SMA but not CD31. In contrast, pharmacological and genetic suppression of autophagy inhibits VSMC migration. Especially, gene silencing of Atg7 inhibited VSMC migration induced by PDGF. Furthermore, three weeks after ligation, markedly decreased neointimal formation was found in mice treated with chloroquine, an inhibitor of autophagy. Quantitative morphometric analysis of the injured vessels revealed a marked reduction in the intima/media ratio in the mice treated with chloroquine. Conclusion: Autophagy activation increases VSMC migration while autophagy suppression inhibits VSMC migration. These findings suggest that autophagy suppression may be an important therapeutic strategy for atherosclerosis and intimal hyperplasia.

Keywords: autophagy, vascular smooth muscle cell, migration, neointimal formation

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132 Surveillance of Artemisinin Resistance Markers and Their Impact on Treatment Outcomes in Malaria Patients in an Endemic Area of South-Western Nigeria

Authors: Abiodun Amusan, Olugbenga Akinola, Kazeem Akano, María Hernández-Castañeda, Jenna Dick, Akintunde Sowunmi, Geoffrey Hart, Grace Gbotosho

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Introduction: Artemisinin-based Combination Therapy (ACTs) is the cornerstone malaria treatment option in most malaria-endemic countries. Unfortunately, the malaria control effort is constantly being threatened by resistance of Plasmodium falciparum to ACTs. The recent evidence of artemisinin resistance in East Africa and its possibility of spreading to other African regions portends an imminent health catastrophe. This study aimed at evaluating the occurrence, prevalence, and influence of artemisinin-resistance markers on treatment outcomes in Ibadan before and after post-adoption of artemisinin combination therapy (ACTs) in Nigeria in 2005. Method: The study involved day zero dry blood spot (DBS) obtained from malaria patients during retrospective (2000-2005) and prospective (2021) studies. A cohort in the prospective study received oral dihydroartemisinin-piperaquine and underwent a 42-day follow-up to observe treatment outcomes. Genomic DNA was extracted from the DBS samples using a QIAamp blood extraction kit. Fragments of P. falciparum kelch13 (Pfkelch13), P. falciparum coronin (Pfcoronin), P. falciparum multidrug resistance 2 (PfMDR2), and P. falciparum chloroquine resistance transporter (PfCRT) genes were amplified and sequenced on a sanger sequencing platform to identify artemisinin resistance-associated mutations. Mutations were identified by aligning sequenced data with reference sequences obtained from the National Center for Biotechnology Information. Data were analyzed using descriptive statistics and student t-tests. Results: Mean parasite clearance time (PCT) and fever clearance time (FCT) were 2.1 ± 0.6 days (95% CI: 1.97-2.24) and 1.3 ± 0.7 days (95% CI: 1.1-1.6) respectively. Four mutations, K189T [34/53(64.2%)], R255K [2/53(3.8%)], K189N [1/53(1.9%)] and N217H [1/53(1.9%)] were identified within the N-terminal (Coiled-coil containing) domain of Pfkelch13. No artemisinin resistance-associated mutation usually found within the β-propeller domain of the Pfkelch13 gene was found in these analyzed samples. However, K189T and R255K mutations showed a significant correlation with longer parasite clearance time in the patients (P<0.002). The observed Pfkelch13 gene changes did not influence the baseline mean parasitemia (P = 0.44). P76S [17/100 (17%)] and V62M [1/100 (1%)] changes were identified in the Pfcoronin gene fragment without any influence on the parasitological parameters. No change was observed in the PfMDR2 gene, while no artemisinin resistance-associated mutation was found in the PfCRT gene. Furthermore, a sample each in the retrospective study contained the Pfkelch13 K189T and Pfcoronin P76S mutations. Conclusion: The study revealed absence of genetic-based evidence of artemisinin resistance in the study population at the time of study. The high frequency of K189T Pfkelch13 mutation and its correlation with increased parasite clearance time in this study may depict geographical variation of resistance mediators and imminent artemisinin resistance, respectively. The study also revealed an inherent potential of parasites to harbour drug-resistant genotypes before the introduction of ACTs in Nigeria.

Keywords: artemisinin resistance, plasmodium falciparum, Pfkelch13 mutations, Pfcoronin

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131 Estimation of State of Charge, State of Health and Power Status for the Li-Ion Battery On-Board Vehicle

Authors: S. Sabatino, V. Calderaro, V. Galdi, G. Graber, L. Ippolito

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Climate change is a rapidly growing global threat caused mainly by increased emissions of carbon dioxide (CO₂) into the atmosphere. These emissions come from multiple sources, including industry, power generation, and the transport sector. The need to tackle climate change and reduce CO₂ emissions is indisputable. A crucial solution to achieving decarbonization in the transport sector is the adoption of electric vehicles (EVs). These vehicles use lithium (Li-Ion) batteries as an energy source, making them extremely efficient and with low direct emissions. However, Li-Ion batteries are not without problems, including the risk of overheating and performance degradation. To ensure its safety and longevity, it is essential to use a battery management system (BMS). The BMS constantly monitors battery status, adjusts temperature and cell balance, ensuring optimal performance and preventing dangerous situations. From the monitoring carried out, it is also able to optimally manage the battery to increase its life. Among the parameters monitored by the BMS, the main ones are State of Charge (SoC), State of Health (SoH), and State of Power (SoP). The evaluation of these parameters can be carried out in two ways: offline, using benchtop batteries tested in the laboratory, or online, using batteries installed in moving vehicles. Online estimation is the preferred approach, as it relies on capturing real-time data from batteries while operating in real-life situations, such as in everyday EV use. Actual battery usage conditions are highly variable. Moving vehicles are exposed to a wide range of factors, including temperature variations, different driving styles, and complex charge/discharge cycles. This variability is difficult to replicate in a controlled laboratory environment and can greatly affect performance and battery life. Online estimation captures this variety of conditions, providing a more accurate assessment of battery behavior in real-world situations. In this article, a hybrid approach based on a neural network and a statistical method for real-time estimation of SoC, SoH, and SoP parameters of interest is proposed. These parameters are estimated from the analysis of a one-day driving profile of an electric vehicle, assumed to be divided into the following four phases: (i) Partial discharge (SoC 100% - SoC 50%), (ii) Partial discharge (SoC 50% - SoC 80%), (iii) Deep Discharge (SoC 80% - SoC 30%) (iv) Full charge (SoC 30% - SoC 100%). The neural network predicts the values of ohmic resistance and incremental capacity, while the statistical method is used to estimate the parameters of interest. This reduces the complexity of the model and improves its prediction accuracy. The effectiveness of the proposed model is evaluated by analyzing its performance in terms of square mean error (RMSE) and percentage error (MAPE) and comparing it with the reference method found in the literature.

Keywords: electric vehicle, Li-Ion battery, BMS, state-of-charge, state-of-health, state-of-power, artificial neural networks

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130 Electrical Decomposition of Time Series of Power Consumption

Authors: Noura Al Akkari, Aurélie Foucquier, Sylvain Lespinats

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Load monitoring is a management process for energy consumption towards energy savings and energy efficiency. Non Intrusive Load Monitoring (NILM) is one method of load monitoring used for disaggregation purposes. NILM is a technique for identifying individual appliances based on the analysis of the whole residence data retrieved from the main power meter of the house. Our NILM framework starts with data acquisition, followed by data preprocessing, then event detection, feature extraction, then general appliance modeling and identification at the final stage. The event detection stage is a core component of NILM process since event detection techniques lead to the extraction of appliance features. Appliance features are required for the accurate identification of the household devices. In this research work, we aim at developing a new event detection methodology with accurate load disaggregation to extract appliance features. Time-domain features extracted are used for tuning general appliance models for appliance identification and classification steps. We use unsupervised algorithms such as Dynamic Time Warping (DTW). The proposed method relies on detecting areas of operation of each residential appliance based on the power demand. Then, detecting the time at which each selected appliance changes its states. In order to fit with practical existing smart meters capabilities, we work on low sampling data with a frequency of (1/60) Hz. The data is simulated on Load Profile Generator software (LPG), which was not previously taken into consideration for NILM purposes in the literature. LPG is a numerical software that uses behaviour simulation of people inside the house to generate residential energy consumption data. The proposed event detection method targets low consumption loads that are difficult to detect. Also, it facilitates the extraction of specific features used for general appliance modeling. In addition to this, the identification process includes unsupervised techniques such as DTW. To our best knowledge, there exist few unsupervised techniques employed with low sampling data in comparison to the many supervised techniques used for such cases. We extract a power interval at which falls the operation of the selected appliance along with a time vector for the values delimiting the state transitions of the appliance. After this, appliance signatures are formed from extracted power, geometrical and statistical features. Afterwards, those formed signatures are used to tune general model types for appliances identification using unsupervised algorithms. This method is evaluated using both simulated data on LPG and real-time Reference Energy Disaggregation Dataset (REDD). For that, we compute performance metrics using confusion matrix based metrics, considering accuracy, precision, recall and error-rate. The performance analysis of our methodology is then compared with other detection techniques previously used in the literature review, such as detection techniques based on statistical variations and abrupt changes (Variance Sliding Window and Cumulative Sum).

Keywords: electrical disaggregation, DTW, general appliance modeling, event detection

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129 Application of Laser-Induced Breakdown Spectroscopy for the Evaluation of Concrete on the Construction Site and in the Laboratory

Authors: Gerd Wilsch, Tobias Guenther, Tobias Voelker

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In view of the ageing of vital infrastructure facilities, a reliable condition assessment of concrete structures is becoming of increasing interest for asset owners to plan timely and appropriate maintenance and repair interventions. For concrete structures, reinforcement corrosion induced by penetrating chlorides is the dominant deterioration mechanism affecting the serviceability and, eventually, structural performance. The determination of the quantitative chloride ingress is required not only to provide valuable information on the present condition of a structure, but the data obtained can also be used for the prediction of its future development and associated risks. At present, wet chemical analysis of ground concrete samples by a laboratory is the most common test procedure for the determination of the chloride content. As the chloride content is expressed by the mass of the binder, the analysis should involve determination of both the amount of binder and the amount of chloride contained in a concrete sample. This procedure is laborious, time-consuming, and costly. The chloride profile obtained is based on depth intervals of 10 mm. LIBS is an economically viable alternative providing chloride contents at depth intervals of 1 mm or less. It provides two-dimensional maps of quantitative element distributions and can locate spots of higher concentrations like in a crack. The results are correlated directly to the mass of the binder, and it can be applied on-site to deliver instantaneous results for the evaluation of the structure. Examples for the application of the method in the laboratory for the investigation of diffusion and migration of chlorides, sulfates, and alkalis are presented. An example for the visualization of the Li transport in concrete is also shown. These examples show the potential of the method for a fast, reliable, and automated two-dimensional investigation of transport processes. Due to the better spatial resolution, more accurate input parameters for model calculations are determined. By the simultaneous detection of elements such as carbon, chlorine, sodium, and potassium, the mutual influence of the different processes can be determined in only one measurement. Furthermore, the application of a mobile LIBS system in a parking garage is demonstrated. It uses a diode-pumped low energy laser (3 mJ, 1.5 ns, 100 Hz) and a compact NIR spectrometer. A portable scanner allows a two-dimensional quantitative element mapping. Results show the quantitative chloride analysis on wall and floor surfaces. To determine the 2-D distribution of harmful elements (Cl, C), concrete cores were drilled, split, and analyzed directly on-site. Results obtained were compared and verified with laboratory measurements. The results presented show that the LIBS method is a valuable addition to the standard procedures - the wet chemical analysis of ground concrete samples. Currently, work is underway to develop a technical code of practice for the application of the method for the determination of chloride concentration in concrete.

Keywords: chemical analysis, concrete, LIBS, spectroscopy

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128 4D Monitoring of Subsurface Conditions in Concrete Infrastructure Prior to Failure Using Ground Penetrating Radar

Authors: Lee Tasker, Ali Karrech, Jeffrey Shragge, Matthew Josh

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Monitoring for the deterioration of concrete infrastructure is an important assessment tool for an engineer and difficulties can be experienced with monitoring for deterioration within an infrastructure. If a failure crack, or fluid seepage through such a crack, is observed from the surface often the source location of the deterioration is not known. Geophysical methods are used to assist engineers with assessing the subsurface conditions of materials. Techniques such as Ground Penetrating Radar (GPR) provide information on the location of buried infrastructure such as pipes and conduits, positions of reinforcements within concrete blocks, and regions of voids/cavities behind tunnel lining. This experiment underlines the application of GPR as an infrastructure-monitoring tool to highlight and monitor regions of possible deterioration within a concrete test wall due to an increase in the generation of fractures; in particular, during a time period of applied load to a concrete wall up to and including structural failure. A three-point load was applied to a concrete test wall of dimensions 1700 x 600 x 300 mm³ in increments of 10 kN, until the wall structurally failed at 107.6 kN. At each increment of applied load, the load was kept constant and the wall was scanned using GPR along profile lines across the wall surface. The measured radar amplitude responses of the GPR profiles, at each applied load interval, were reconstructed into depth-slice grids and presented at fixed depth-slice intervals. The corresponding depth-slices were subtracted from each data set to compare the radar amplitude response between datasets and monitor for changes in the radar amplitude response. At lower values of applied load (i.e., 0-60 kN), few changes were observed in the difference of radar amplitude responses between data sets. At higher values of applied load (i.e., 100 kN), closer to structural failure, larger differences in radar amplitude response between data sets were highlighted in the GPR data; up to 300% increase in radar amplitude response at some locations between the 0 kN and 100 kN radar datasets. Distinct regions were observed in the 100 kN difference dataset (i.e., 100 kN-0 kN) close to the location of the final failure crack. The key regions observed were a conical feature located between approximately 3.0-12.0 cm depth from surface and a vertical linear feature located approximately 12.1-21.0 cm depth from surface. These key regions have been interpreted as locations exhibiting an increased change in pore-space due to increased mechanical loading, or locations displaying an increase in volume of micro-cracks, or locations showing the development of a larger macro-crack. The experiment showed that GPR is a useful geophysical monitoring tool to assist engineers with highlighting and monitoring regions of large changes of radar amplitude response that may be associated with locations of significant internal structural change (e.g. crack development). GPR is a non-destructive technique that is fast to deploy in a production setting. GPR can assist with reducing risk and costs in future infrastructure maintenance programs by highlighting and monitoring locations within the structure exhibiting large changes in radar amplitude over calendar-time.

Keywords: 4D GPR, engineering geophysics, ground penetrating radar, infrastructure monitoring

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127 Fighting the Crisis with 4.0 Competences: Higher Education Projects in the Times of Pandemic

Authors: Jadwiga Fila, Mateusz Jezowski, Pawel Poszytek

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The outbreak of the global COVID-19 pandemic started the times of crisis full of uncertainty, especially in the field of transnational cooperation projects based on the international mobility of their participants. This is notably the case of Erasmus+ Program for higher education, which is the flagship European initiative boosting cooperation between educational institutions, businesses, and other actors, enabling students and staff mobility, as well as strategic partnerships between different parties. The aim of this abstract is to study whether competences 4.0 are able to empower Erasmus+ project leaders in sustaining their international cooperation in times of global crisis, widespread online learning, and common project disruption or cancellation. The concept of competences 4.0 emerged from the notion of the industry 4.0, and it relates to skills that are fundamental for the current labor market. For the aim of the study presented in this abstract, four main 4.0 competences were distinguished: digital, managerial, social, and cognitive competence. The hypothesis for the study stipulated that the above-mentioned highly-developed competences may act as a protective shield against the pandemic challenges in terms of projects’ sustainability and continuation. The objective of the research was to assess to what extent individual competences are useful in managing projects in times of crisis. For this purpose, the study was conducted, involving, among others, 141 Polish higher education project leaders who were running their cooperation projects during the peak of the COVID-19 pandemic (Mar-Nov 2020). The research explored the self-perception of the above-mentioned competences among Erasmus+ project leaders and the contextual data regarding the sustainability of the projects. The quantitative character of data permitted validation of scales (Cronbach’s Alfa measure), and the use of factor analysis made it possible to create a distinctive variable for each competence and its dimensions. Finally, logistic regression was used to examine the association of competences and other factors on project status. The study shows that the project leaders’ competence profile attributed the highest score to digital competence (4.36 on the 1-5 scale). Slightly lower values were obtained for cognitive competence (3.96) and managerial competence (3.82). The lowest score was accorded to one specific dimension of social competence: adaptability and ability to manage stress (1.74), which proves that the pandemic was a real challenge which had to be faced by project coordinators. For higher education projects, 10% were suspended or prolonged because of the COVID-19 pandemic, whereas 90% were undisrupted (continued or already successfully finished). The quantitative analysis showed a positive relationship between the leaders’ levels of competences and the projects status. In the case of all competences, the scores were higher for project leaders who finished projects successfully than for leaders who suspended or prolonged their projects. The research demonstrated that, in the demanding times of the COVID-19 pandemic, competences 4.0, to a certain extent, do play a significant role in the successful management of Erasmus+ projects. The implementation and sustainability of international educational projects, despite mobility and sanitary obstacles, depended, among other factors, on the level of leaders’ competences.

Keywords: Competences 4.0, COVID-19 pandemic, Erasmus+ Program, international education, project sustainability

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126 Diversification of Productivity of the Oxfordian Subtidal Carbonate Factory in the Holy Cross Mountains

Authors: Radoslaw Lukasz Staniszewski

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The aim of the research was to verify lateral extent and thickness variability of individual limestone layers within early-Jurassic medium- and thick-bedded limestone interbedded with marlstones. Location: The main research area is located in the south-central part of Poland in the south-western part of Permo-Mesozoic margin of the Holy Cross Mountains. It includes outcroppings located on the line between Mieczyn and Wola Morawicka. The analyses were carried out on six profiles (Mieczyn, Gniezdziska, Tokarnia, Wola Morawicka, Morawica and Wolica) representing three early-Jurassic links: Jasna Gora layers, grey limestone, Morawica limestone. Additionally, an attempt was made to correlate the thickness sequence from the Holy Cross Mountains to the profile from the quarry in Zawodzie located 3 km east of Czestochowa. The distance between the outermost profiles is 122 km in a straight line. Methodology of research: The Callovian-Oxfordian border was taken as the reference point during the correlation. At the same time, ammonite-based stratigraphic studies were carried out, which allowed to identify individual packages in the remote outcroppings. The analysis of data collected during fieldwork was mainly devoted to the correlation of thickness sequences of limestone layers in subsequent profiles. In order to check the objectivity of the subsequent outcroppings, the profiles have been presented in the form of the thickness functions of the subsequent layers. The generated functions were auto-correlated, and the Pearson correlation coefficient was calculated. The next step in the research was to statistically determine the percentage increment of the individual layers thickness in the subsequent profiles, and on this basis to plot the function of relative carbonate productivity. Results: The result of the above-mentioned procedures consists in illustrating the extent of 34 rock layers across the examined area in demonstrating the repeatability of their success in subsequent outcroppings. It can also be observed that the thickness of individual layers in the Holy Cross Mountains is increasing from north-west towards south-east. Despite changes in the thickness of the layers in the profiles, their relations within the sequence remain constant. The lowest matching ratio of thickness sequence calculated using the Pearson correlation coefficient formula is 0.67, while the highest is 0.84. The thickness of individual layers changes between 4% and 230% over the examined area. Interpretation: Layers in the outcroppings covered by the research show continuity throughout the examined area and it is possible to precisely correlate them, which means that the process determining the formation of the layers was regional and probably included both the fringe of the Holy Cross Mountains and the north-eastern part of the Krakow-Czestochowa Jura Upland. Local changes in the sedimentation environment affecting the productivity of the subtidal carbonate factory only cause the thickness of the layers to change without altering the thickness proportions of the profiles. Based on the percentage of changes in the thickness of individual layers in the subsequent profiles, it can be concluded that the local productivity of the subtidal carbonate factory is increasing logarithmically.

Keywords: Oxfordian, Holy Cross Mountains, carbonate factory, Limestone

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