Search results for: I. Ferguson

Authors: Moustafa Elwan, Sohier Shehata, Fatma Sedek, Manar Hussine

Abstract:

One of the most risky factors that lead to a foot injury during physical activities are both high and low arched feet. Normally the medial longitudinal arch of the foot develops in the first 10 years of life, so flexible flat foot has an inversely relationship with age in the first decade, all over the world, the prevalence of flat foot is increasing. In approximately 15% of foot deformities cases, the deformity does not disappear and remains throughout adulthood, 90% of the clinical cases are complaining from foot problems are due to flatfoot. Flatfoot creates subtalar over pronation, which creates tibial and femoral medial rotation, and that is accompanied with increases of pelvic tilting anteriorly, which may influence the lumbar vertebrae alignment by increasing muscle tension and rotation. Objective: To study the impact of the flexible flatfoot on lumbosacral angle (angle of Ferguson). Methods: This experiment included 40 volunteers (14 females &26 males) gathered from the Faculty of Physical Therapy, Modern University of Technology and Information, Cairo, Egypt, for each participant, four angles were measured in the foot( talar first metatarsal angle, lateral talocalcaneal angle, , Calcaneal first metatarsal angle, calcaneal inclination angle) and one angle in the lumbar region (lumbosacral angle). Measurement of these angles was conducted by using Surgimap Spine software (version 2.2.9.6). Results: The results demonstrated that there was no significant correlation betweenFerguson angle and lateral talocalcaneal (r=0.164, p=0.313). Also, there was no significant correlation between Ferguson angle and talo first metatarsal “Meary’s angle" (r=0.007, p=0.968). Moreover, there was no significant correlation between Ferguson angle and calcaneal-first metatarsal angle (r=0.083, p=0.612). Also, there was no significant correlation between Ferguson angle and calcaneal inclination angle (r= 0.032, p= 0.846). Conclusion: It can be concluded that there is no significant correlation between the flexible flat foot and lumbosacral angle So, more study should be conducted in large sample and different ages and conditions of foot problems.

Keywords: calcaneal first metatarsal, calcaneal inclination, flatfoot, ferguson’s angle, lateral talocalcaneal angle, lumbosacral angle, and talar first metatarsal angle

Procedia PDF Downloads 132

Authors: Nidal F. Shilbayeh, Reem A. Al-Saidi, Ahmed H. Alsswey

Abstract:

In this paper, we presented an evaluation and analysis of E-Voting Authentication Preparation Scheme (EV-APS). EV-APS applies some modified security aspects that enhance the security measures and adds a strong wall of protection, confidentiality, non-repudiation and authentication requirements. Some of these modified security aspects are Kerberos authentication protocol, PVID scheme, responder certificate validation, and the converted Ferguson e-cash protocol. Authentication and privacy requirements have been evaluated and proved. Authentication guaranteed only eligible and authorized voters were permitted to vote. Also, the privacy guaranteed that all votes will be kept secret. Evaluation and analysis of some of these security requirements have been given. These modified aspects will help in filtering the counter buffer from unauthorized votes by ensuring that only authorized voters are permitted to vote.

Keywords: e-voting preparation stage, blind signature protocol, Nonce based authentication scheme, Kerberos Authentication Protocol, pseudo voter identity scheme PVID

Procedia PDF Downloads 297

Authors: Brian Kozak, Ryan Ferguson, Evan Hockeridge

Abstract:

Aerospace engineering and aeronautical engineering students studying propulsion often learn about propellers and their importance in aviation propulsion. In order to reinforce concepts introduced in the classroom, laboratory projects are used. However, to test a full scale propeller, an engine mounted on a test stand must be used. This engine needs to be enclosed in a test cell for appropriated safety requirements, is expensive to operate, and requires a significant amount of time to change propellers. In order to decrease costs and time requirements, the authors designed and built an electric motor powered desktop Arduino controlled test stand. This test stand is used to enhance student understanding of propeller size and pitch on thrust. The test stand can accommodate propellers up to 25 centimeters in diameter. The code computer allowed for the motor speed to be increased or decreased by 1% per second. Outputs that are measured are thrust, motor rpm, amperes, voltage, and motor temperature. These data are exported as a .CVS file and can be imported into a graphing program for data analysis.

Keywords: Arduino, Laboratory Project, Test stand, Propeller

Procedia PDF Downloads 219

Authors: M. Asghar, K. Mahmood, F. Malik, Lu Na, Y-H Xie, Yasin A. Raja, I. Ferguson

Abstract:

In this paper, we have reported an enhancement in Seebeck coefficient of un-doped zinc oxide (ZnO) grown by molecular beam epitaxy (MBE) on silicon (001) substrate by annealing treatment. The grown ZnO thin films were annealed in oxygen environment at 500°C – 800°C, keeping a step of 100°C for one hour. Room temperature Seebeck measurements showed that Seebeck coefficient and power factor increased from 222 to 510 µV/K and 8.8×10^-6 to 2.6×10^-4 Wm^-1K^-2 as annealing temperature increased from 500°C to 800°C respectively. This is the highest value of Seebeck coefficient ever reported for un-doped MBE grown ZnO according to best of our knowledge. This observation was related with the improvement of crystal structure of grown films with annealing temperature. X-ray diffraction (XRD) results demonstrated that full width half maximum (FWHM) of ZnO (002) plane decreased and crystalline size increased as the annealing temperature increased. Photoluminescence study revealed that the intensity of band edge emission increased and defect emission decreased as annealing temperature increased because the density of oxygen vacancy related donor defects decreased with annealing temperature. This argument was further justified by the Hall measurements which showed a decreasing trend of carrier concentration with annealing temperature.

Keywords: ZnO, MBE, thermoelectric properties, annealing temperature, crystal structure

Procedia PDF Downloads 445

Authors: G. Culache, M. P. Byfield, N. S. Ferguson, A. Tyas

Abstract:

Steel joints in building structures represent a weak link in the case of accidental transient loading. This type of loading can occur due to blast effects or impact with moving vehicles and will result in large deformations in the material as well as large rotations. This paper addresses the lack of experimental investigations into the response of moment resisting connections subjected to such loading. The current design philosophy was used to create test specimens with flush and extended end plates. The specimens were tested in a specially designed testing rig capable of delivering the sustained loading even beyond the point of failure. Types of failure that the authors attempted to obtain were bolt fracture, flange crushing and end plate fracture. Experimental data is presented, described and analyzed. The tests show that the strength and ductility can be significantly improved by replacing ordinary mild-steel bolts with their stainless steel equivalents. This minor modification is demonstrated to significantly improve the robustness when subjected to loading that results in high deformations and rotation, where loading is maintained during failure. Conclusions are drawn about the wider implications of this research and recommendations made on the direction of future research in this field.

Keywords: steel moment connections, high strain rates, dynamic loading, experimental testing

Procedia PDF Downloads 321

Authors: Peter Ferguson

Abstract:

In 2018, Japan’s Ministry of Education revised the public elementary school curriculum. As part of widespread reforms, the recent Course of Study established English as an academic subject in Grades 5 and 6 plus lowered the starting age of 'foreign language activities' to Grade 3. These changes were implemented in April 2020. This presentation will examine the process and effects that policy implementation had on schools and teachers. A critical analysis of the 2018 Course of Study policy documents revealed several discourses were expressed concerning not only English education and foreign language acquisition, but that larger political and socioeconomic ideological beliefs on globalization, language, nation, culture, and identity were also articulated. Using excerpts from document analysis, the presenter will demonstrate how competing discourses were expressed in policy texts. Data from interviews with national policymakers also exposed several challenges policymakers faced as they tried to balance competing discourses and articulate important pedagogical concepts while having their voices heard. Findings show that some stakeholders were marginalized during the processes of policy creation, transmission, and implementation. This presentation is part of a larger multiple case study that utilized ethnography of language policy and critical analysis of discourse to examine how English education language policy was implemented into the national elementary school curriculum in Japan, and how stakeholders at the various educational levels contended with the creation, interpretation, and appropriation of the language policy.

Keywords: ethnography of language policy, elementary school EFL, language ideologies, discourse analysis

Procedia PDF Downloads 118

Authors: Fumio Kasai, Noriko Hirayama, Jorge Pereira, Azusa Ohtani, Masashi Iemura, Malcolm A. Ferguson Smith, Arihiro Kohara

Abstract:

The VERO cell line was established in 1962 from normal tissue of an African green monkey, Chlorocebus aethiops (2n=60), and has been commonly used worldwide for screening for toxins or as a cell substrate for the production of viral vaccines. The VERO genome was sequenced in 2014; however, its cytogenetic features have not been fully characterized as it contains several chromosome abnormalities and different karyotypes coexist in the cell line. In this study, the VERO cell line (JCRB0111) was compared with one of the sublines. In contrast to 59 chromosomes as the modal chromosome number in the VERO cell line, the subline had two peaks of 56 and 58 chromosomes. M-FISH analysis using human probes revealed that the VERO cell line was characterized by a translocation t(2;25) found in all metaphases, which was absent in the subline. Different abnormalities detected only in the subline show that the cell line is heterogeneous, indicating that the subline has the potential to change its genomic characteristics during cell culture. The various alterations in the two independent lineages suggest that genomic changes in both VERO cells can be accounted for by progressive rearrangements during their evolution in culture. Both t(5;X) and t(8;14) observed in all metaphases of the two cell lines might have a key role in VERO cells and could be used as genetic markers to identify VERO cells. The flow karyotype shows distinct differences from normal. Further analysis of sorted abnormal chromosomes may uncover other characteristics of VERO cells. Because of the absence of STR data, cytogenetic data are important in characterizing animal cell lines and can be an indicator of their quality control.

Keywords: VERO, cell culture passage, chromosome rearrangement, heterogeneous cells

Procedia PDF Downloads 416

Authors: Marcya Burden, Savonna Greer

Abstract:

In the wake of social unrest around the world, protest movements have been captured like never before. As protest movements have evolved, so too have their visibility and sources of coverage. Long gone are the days of print media as our only glimpse into the action surrounding a protest. Now, with social networks such as Facebook, Instagram and Snapchat, we have access to real-time video footage of protest movements and human rights activism that can reach millions of people within seconds. This research paper investigated various social media network platforms’ statistical usage data in the areas of human rights activism and protest movements, paralleling with other past forms of media coverage. This research demonstrates that social networks are extremely important to protest movements and human rights activism. With over 2.9 billion users across social media networks globally, these platforms are the heart of most recent protests and human rights activism. This research shows the paradigm shift from the Selma March of 1965 to the more recent protests of Ferguson in 2014, Ni Una Menos in 2015, and End Sars in 2018. The research findings demonstrate that today, almost anyone may use their social networks to protest movement leaders and human rights activists. From a student to an 80-year-old professor, the possibility of reaching billions of people all over the world is limitless. Findings show that 82% of the world’s internet population is on social networks 1 in every 5 minutes. Over 65% of Americans believe social media highlights important issues. Thus, there is no need to have a formalized group of people or even be known online. A person simply needs to be engaged on their respective social media networks (Facebook, Twitter, Instagram, Snapchat) regarding any cause they are passionate about. Information may be exchanged in real time around the world and a successful protest can begin.

Keywords: activism, protests, human rights, networks

Procedia PDF Downloads 95

Authors: Matthew Ferguson, Tatyana Konkova, Ioannis Violatos

Abstract:

Microstructure analysis of additively manufactured (AM) materials is an important step in understanding the interrelationship between mechanical properties and materials performance. Literature on the effect of laser-based AM process parameters on the microstructure in the substrate-deposit interface is limited. The interface region, the adjoining area of substrate and deposit, is characterized by the presence of the fusion zone (FZ) and heat-affected zone (HAZ), experiencing rapid thermal gyrations resulting in thermal-induced transformations. Inconel 718 was utilized as work material for both the substrate and deposit. Three blocks of Inconel 718 material were deposited by Direct Energy Deposition (DED) using three different laser powers, 550W, 750W and 950W, respectively. A coupled thermo-mechanical transient approach was utilized to correlate temperature history to the evolution of microstructure. The thermal history of the deposition process was monitored with the thermocouples installed inside the substrate material. The interface region of the blocks was analyzed with Optical Microscopy (OM) and Scanning Electron Microscopy (SEM), including the electron back-scattered diffraction (EBSD) technique. Laser power was found to influence the dissolution of intermetallic precipitated phases in the substrate and grain growth in the interface region. Microstructure and thermal history data were utilized to draw conclusive comparisons between the investigated process parameters.

Keywords: additive manufacturing, direct energy deposition, electron back-scattered diffraction, finite element analysis, inconel 718, microstructure, optical microscopy, scanning electron microscopy, substrate-deposit interface region

Procedia PDF Downloads 201

Authors: Khatuna Buskivadze

Abstract:

The aim of the present study is to investigate the sociolinguistic and classroom functions and frequency of Teacher’s Code Switching (CS) in the Content and Language Integrated (CLIL) Lesson. Nowadays, Georgian society struggles to become the part of the European world, the English language itself plays a role in forming new generations with European values. Based on our research conducted in 2019, out of all 114 private schools in Tbilisi, full- programs of CLIL are taught in 7 schools, while only some subjects using CLIL are conducted in 3 schools. The goal of the former research was to define the features of Content and Language Integrated learning (CLIL) methodology within the process of teaching English on the Example of Georgian private high schools. Taking the Georgian reality and cultural features into account, the modified version of the questionnaire, based on the classification of using CS in ESL Classroom proposed By Ferguson (2009) was used. The qualitative research revealed students’ and teacher’s attitudes towards teacher’s code-switching in CLIL lesson. Both qualitative and quantitative research were conducted: the observations of the teacher’s lessons (Recording of T’s online lessons), interview and the questionnaire among Math’s T’s 20 high school students. We came to the several conclusions, some of them are given here: Math’s teacher’s CS behavior mostly serves (1) the conversational function of interjection; (2) the classroom functions of introducing unfamiliar materials and topics, explaining difficult concepts, maintaining classroom discipline and the structure of the lesson; The teacher and 13 students have negative attitudes towards using only Georgian in teaching Math. The higher level of English is the more negative is attitude towards using Georgian in the classroom. Although all the students were Georgian, their competence in English is higher than in Georgian, therefore they consider English as an inseparable part of their identities. The overall results of the case study of teaching Math (Educational discourse) in one of the private schools in Tbilisi will be presented at the conference.

Keywords: attitudes, bilingualism, code-switching, CLIL, conversation analysis, interactional sociolinguistics.

Procedia PDF Downloads 160

Authors: Robin Ferguson

Abstract:

The context which this paper explores is the provision of Technical Vocational Education and Training (TVET) of southern African workers at sea on local and foreign vessels using a blended learning approach. The pedagogical challenge of providing quality education in this context is that multiple African and foreign languages and cultural norms are found amongst the all-male crew; and there are widely differing levels of education, low levels of digital literacy and limited connectivity. The methodology used is a nested case study. The study describes the mechanisms used to provide ongoing, real-time workplace TVET on two foreign vessels. Some training was done in person when the vessels came into port, however, the majority of the TVET was achieved from shore to ship using a combination of commonly available Google and Microsoft Apps and WhatsApp. Voice, video and text in multiple languages were used to accommodate different learning styles. The learning was supported by the development of learning networks using social media. This paper also reflects on the shore-based organisational change processes required to support sea learning. The conceptual framework used is the Theory of Practice Architectures (TPA) as is provides a site-ontological perspective of the sayings/thinkings, doings and relatings of this workplace training which is multiplanar as it plays out at sea and ashore, in-person and on-line. Using TPA, the overarching practice architectures and supporting structures which confound or enable these learning practices are revealed. The contribution which this paper makes is an insight into an innovative vocational pedagogy which promotes ICT-mediated learning amongst workers who suffer from low levels of literacies and limited ICT-access and who work and live in remote places. It is a pedagogy of possibility which crosses the digital divide.

Keywords: theory of practice architecture, microsoft, google, whatsapp, vocational pedagogy, mariners, distributed workplaces

Procedia PDF Downloads 80

Authors: Rajinder Singh, Ram Valluru

Abstract:

Chain Ladder (CL) method, Expected Loss Ratio (ELR) method and Bornhuetter-Ferguson (BF) method, in addition to more complex transition-rate modeling, are commonly used actuarial reserving methods in general insurance. There is limited published research about their relative performance in the context of Mortgage Insurance (MI). In our experience, these traditional techniques pose unique challenges and do not provide stable claim estimates for medium to longer term liabilities. The relative strengths and weaknesses among various alternative approaches revolve around: stability in the recent loss development pattern, sufficiency and reliability of loss development data, and agreement/disagreement between reported losses to date and ultimate loss estimate. CL method results in volatile reserve estimates, especially for accident periods with little development experience. The ELR method breaks down especially when ultimate loss ratios are not stable and predictable. While the BF method provides a good tradeoff between the loss development approach (CL) and ELR, the approach generates claim development and ultimate reserves that are disconnected from the ever-to-date (ETD) development experience for some accident years that have more development experience. Further, BF is based on subjective a priori assumption. The fundamental shortcoming of these methods is their inability to model exogenous factors, like the economy, which impact various cohorts at the same chronological time but at staggered points along their life-time development. This paper proposes an alternative approach of parametrizing the loss development curve and using logistic regression to generate the ultimate loss estimate for each homogeneous group (accident year or delinquency period). The methodology was tested on an actual MI claim development dataset where various cohorts followed a sigmoidal trend, but levels varied substantially depending upon the economic and operational conditions during the development period spanning over many years. The proposed approach provides the ability to indirectly incorporate such exogenous factors and produce more stable loss forecasts for reserving purposes as compared to the traditional CL and BF methods.

Keywords: actuarial loss reserving techniques, logistic regression, parametric function, volatility

Procedia PDF Downloads 130

Authors: Frederick Ferguson, Dehua Feng, Yang Gao

Abstract:

No doubt, current CFD tools have a great many technical limitations, and active research is being done to overcome these limitations. Current areas of limitations include vortex-dominated flows, separated flows, and turbulent flows. In general, turbulent flows are unsteady solutions to the fluid dynamic equations, and instances of these solutions can be computed directly from the equations. One of the approaches commonly implemented is known as the ‘direct numerical simulation’, DNS. This approach requires a spatial grid that is fine enough to capture the smallest length scale of the turbulent fluid motion. This approach is called the ‘Kolmogorov scale’ model. It is of interest to note that the Kolmogorov scale model must be captured throughout the domain of interest and at a correspondingly small-time step. In typical problems of industrial interest, the ratio of the length scale of the domain to the Kolmogorov length scale is so great that the required grid set becomes prohibitively large. As a result, the available computational resources are usually inadequate for DNS related tasks. At this time in its development, DNS is not applicable to industrial problems. In this research, an attempt is made to develop a numerical technique that is capable of delivering DNS quality solutions at the scale required by the industry. To date, this technique has delivered preliminary results for both steady and unsteady, viscous and inviscid, compressible and incompressible, and for both high and low Reynolds number flow fields that are very accurate. Herein, it is proposed that the Integro-Differential Scheme (IDS) be applied to a set of vortex-shockwave interaction problems with the goal of investigating the nonstationary physics within the resulting interaction regions. In the proposed paper, the IDS formulation and its numerical error capability will be described. Further, the IDS will be used to solve the inviscid and viscous Burgers equation, with the goal of analyzing their solutions over a considerable length of time, thus demonstrating the unsteady capabilities of the IDS. Finally, the IDS will be used to solve a set of fluid dynamic problems related to flow that involves highly vortex interactions. Plans are to solve the following problems: the travelling wave and vortex problems over considerable lengths of time, the normal shockwave–vortex interaction problem for low supersonic conditions and the reflected oblique shock–vortex interaction problem. The IDS solutions obtained in each of these solutions will be explored further in efforts to determine the distributed density gradients and vorticity, as well as the Q-criterion. Parametric studies will be conducted to determine the effects of the Mach number on the intensity of vortex-shockwave interactions.

Keywords: vortex dominated flows, shockwave interactions, high Reynolds number, integro-differential scheme

Procedia PDF Downloads 136

Authors: Tuğrul Varol, Halil Barış Özel

Abstract:

In this study which has been conducted in Akçasu Forest Range District of Devrek Forest Directorate; 3 methods (cover removal with human force, cover removal with Hitachi F20 Excavator, and cover removal with agricultural equipment mounted on a Ferguson 240S agriculture tractor) utilized in weed control efforts in regeneration of degraded oriental beech forests have been compared. In this respect, 3 methods have been compared by determining certain work hours and standard durations of unit areas (1 hectare). For this purpose, evaluating the tasks made with human and machine force from the aspects of duration, productivity and costs, it has been aimed to determine the most productive method in accordance with the actual ecological conditions of research field. Within the scope of the study, the time studies have been conducted for 3 methods used in weed control efforts. While carrying out those studies, the performed implementations have been evaluated by dividing them into business stages. Also, the actual data have been used while calculating the cost accounts. In those calculations, the latest formulas and equations which are also used in developed countries have been utilized. The variance of analysis (ANOVA) was used in order to determine whether there is any statistically significant difference among obtained results, and the Duncan test was used for grouping if there is significant difference. According to the measurements and findings carried out within the scope of this study, it has been found during living cover removal efforts in regeneration efforts in demolished oriental beech forests that the removal of weed layer in 1 hectare of field has taken 920 hours with human force, 15.1 hours with excavator and 60 hours with an equipment mounted on a tractor. On the other hand, it has been determined that the cost of removal of living cover in unit area (1 hectare) was 3220.00 TL for man power, 788.70 TL for excavator and 2227.20 TL for equipment mounted on a tractor. According to the obtained results, it has been found that the utilization of excavator in weed control effort in regeneration of degraded oriental beech regions under actual ecological conditions of research field has been found to be more productive from both of aspects of duration and costs. These determinations carried out should be repeated in weed control efforts in degraded forest fields with different ecological conditions, it is compulsory for finding the most efficient weed control method. These findings will light the way of technical staff of forestry directorate in determination of the most effective and economic weed contol method. Thus, the more actual data will be used while preparing the weed control budgets, and there will be significant contributions to national economy. Also the results of this and similar studies are very important for developing the policies for our forestry in short and long term.

Keywords: artificial regeneration, weed control, oriental beech, productivity, mechanization, man power, cost analysis

Procedia PDF Downloads 418

Authors: Jingsi Li, Neil S. Ferguson

Abstract:

Time pressure could have an influence on the productivity, quality of decision making, and the efficiency of problem-solving. This has been mostly stemmed from cognitive research or psychological literature. However, a salient scarce discussion has been held for transport adjacent fields. It is conceivable that in many activity-travel contexts, time pressure is a potentially important factor since an excessive amount of decision time may incur the risk of late arrival to the next activity. The activity-travel rescheduling behavior is commonly explained by costs and benefits of factors such as activity engagements, personal intentions, social requirements, etc. This paper hypothesizes that an additional factor of perceived time pressure could affect travelers’ rescheduling behavior, thus leading to an impact on travel demand management. Time pressure may arise from different ways and is assumed here to be essentially incurred due to travelers planning their schedules without an expectation of unforeseen elements, e.g., transport disruption. In addition to a linear-additive utility-maximization model, the less computationally compensatory heuristic models are considered as an alternative to simulate travelers’ responses. The paper will contribute to travel behavior modeling research by investigating the following questions: how to measure the time pressure properly in an activity-travel day plan context? How do travelers reschedule their plans to cope with the time pressure? How would the importance of the activity affect travelers’ rescheduling behavior? What will the behavioral model be identified to describe the process of making activity-travel rescheduling decisions? How do these identified coping strategies affect the transport network? In this paper, a Mixed Heuristic Model (MHM) is employed to identify the presence of different choice heuristics through a latent class approach. The data about travelers’ activity-travel rescheduling behavior is collected via a web-based interactive survey where a fictitious scenario is created comprising multiple uncertain events on the activity or travel. The experiments are conducted in order to gain a real picture of activity-travel reschedule, considering the factor of time pressure. The identified behavioral models are then integrated into a multi-agent transport simulation model to investigate the effect of the rescheduling strategy on the transport network. The results show that an increased proportion of travelers use simpler, non-compensatory choice strategies instead of compensatory methods to cope with time pressure. Specifically, satisfying - one of the heuristic decision-making strategies - is adopted commonly since travelers tend to abandon the less important activities and keep the important ones. Furthermore, the importance of the activity is found to increase the weight of negative information when making trip-related decisions, especially route choices. When incorporating the identified non-compensatory decision-making heuristic models into the agent-based transport model, the simulation results imply that neglecting the effect of perceived time pressure may result in an inaccurate forecast of choice probability and overestimate the affectability to the policy changes.

Keywords: activity-travel rescheduling, decision making under uncertainty, mixed heuristic model, perceived time pressure, travel demand management

Procedia PDF Downloads 112

Authors: Judy M. Obliosca, Olivia Vest, Sandra Poulos, Kelsi Smith, Tammy Ferguson, Abigail Powers Lott, Alicia K. Smith, Yang Xu, Christopher K. Tison

Abstract:

Post-Traumatic Stress Disorder (PTSD) is a mental health problem that people may develop after experiencing traumatic events such as combat, natural disasters, and major emotional challenges. Tragically, the number of military personnel with PTSD correlates directly with the number of veterans who attempt suicide, with the highest rate in the Army. Research has shown epigenetic risks in those who are prone to several psychiatric dysfunctions, particularly PTSD. Once initiated in response to trauma, epigenetic alterations in particular, the DNA methylation in the form of 5-methylcytosine (5mC) alters chromatin structure and represses gene expression. Current methods to detect DNA methylation, such as bisulfite-based genomic sequencing techniques, are laborious and have massive analysis workflow while still having high error rates. A faster and simpler detection method of high sensitivity and precision would be useful in a clinical setting to confirm potential PTSD etiologies, prevent other psychiatric disorders, and improve military health. A nano-enhanced Surface Plasmon Resonance imaging (SPRi)-based assay that simultaneously detects site-specific 5mC base (termed as PTSD base) in methylated genes related to PTSD is being developed. The arrays on a sensing chip were first constructed for parallel detection of PTSD bases using synthetic and genomic DNA (gDNA) samples. For the gDNA sample extracted from the whole blood of a PTSD patient, the sample was first digested using specific restriction enzymes, and fragments were denatured to obtain single-stranded methylated target genes (ssDNA). The resulting mixture of ssDNA was then injected into the assay platform, where targets were captured by specific DNA aptamer probes previously immobilized on the surface of a sensing chip. The PTSD bases in targets were detected by anti-5-methylcytosine antibody (anti-5mC), and the resulting signals were then enhanced by the universal nanoenhancer. Preliminary results showed successful detection of a PTSD base in a gDNA sample. Brighter spot images and higher delta values (control-subtracted reflectivity signal) relative to those of the control were observed. We also implemented the in-house surface activation system for detection and developed SPRi disposable chips. Multiplexed PTSD base detection of target methylated genes in blood DNA from PTSD patients of severity conditions (asymptomatic and severe) was conducted. This diagnostic capability being developed is a platform technology, and upon successful implementation for PTSD, it could be reconfigured for the study of a wide variety of neurological disorders such as traumatic brain injury, Alzheimer’s disease, schizophrenia, and Huntington's disease and can be extended to the analyses of other sample matrices such as urine and saliva.

Keywords: epigenetic assay, DNA methylation, PTSD, whole blood, multiplexing

Procedia PDF Downloads 121