Search results for: DNA damage response
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7361

Search results for: DNA damage response

5741 Understanding the Fundamental Driver of Semiconductor Radiation Tolerance with Experiment and Theory

Authors: Julie V. Logan, Preston T. Webster, Kevin B. Woller, Christian P. Morath, Michael P. Short

Abstract:

Semiconductors, as the base of critical electronic systems, are exposed to damaging radiation while operating in space, nuclear reactors, and particle accelerator environments. What innate property allows some semiconductors to sustain little damage while others accumulate defects rapidly with dose is, at present, poorly understood. This limits the extent to which radiation tolerance can be implemented as a design criterion. To address this problem of determining the driver of semiconductor radiation tolerance, the first step is to generate a dataset of the relative radiation tolerance of a large range of semiconductors (exposed to the same radiation damage and characterized in the same way). To accomplish this, Rutherford backscatter channeling experiments are used to compare the displaced lattice atom buildup in InAs, InP, GaP, GaN, ZnO, MgO, and Si as a function of step-wise alpha particle dose. With this experimental information on radiation-induced incorporation of interstitial defects in hand, hybrid density functional theory electron densities (and their derived quantities) are calculated, and their gradient and Laplacian are evaluated to obtain key fundamental information about the interactions in each material. It is shown that simple, undifferentiated values (which are typically used to describe bond strength) are insufficient to predict radiation tolerance. Instead, the curvature of the electron density at bond critical points provides a measure of radiation tolerance consistent with the experimental results obtained. This curvature and associated forces surrounding bond critical points disfavors localization of displaced lattice atoms at these points, favoring their diffusion toward perfect lattice positions. With this criterion to predict radiation tolerance, simple density functional theory simulations can be conducted on potential new materials to gain insight into how they may operate in demanding high radiation environments.

Keywords: density functional theory, GaN, GaP, InAs, InP, MgO, radiation tolerance, rutherford backscatter channeling

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5740 Impact Deformation and Fracture Behaviour of Cobalt-Based Haynes 188 Superalloy

Authors: Woei-Shyan Lee, Hao-Chien Kao

Abstract:

The impact deformation and fracture behaviour of cobalt-based Haynes 188 superalloy are investigated by means of a split Hopkinson pressure bar. Impact tests are performed at strain rates ranging from 1×103 s-1 to 5×103 s-1 and temperatures between 25°C and 800°C. The experimental results indicate that the flow response and fracture characteristics of cobalt-based Haynes 188 superalloy are significantly dependent on the strain rate and temperature. The flow stress, work hardening rate and strain rate sensitivity all increase with increasing strain rate or decreasing temperature. It is shown that the impact response of the Haynes 188 specimens is adequately described by the Zerilli-Armstrong fcc model. The fracture analysis results indicate that the Haynes 188 specimens fail predominantly as the result of intensive localised shearing. Furthermore, it is shown that the flow localisation effect leads to the formation of adiabatic shear bands. The fracture surfaces of the deformed Haynes 188 specimens are characterised by dimple- and / or cleavage-like structure with knobby features. The knobby features are thought to be the result of a rise in the local temperature to a value greater than the melting point.

Keywords: Haynes 188 alloy, impact, strain rate and temperature effect, adiabatic shearing

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5739 The Design of Intelligent Passenger Organization System for Metro Stations Based on Anylogic

Authors: Cheng Zeng, Xia Luo

Abstract:

Passenger organization has always been an essential part of China's metro operation and management. Facing the massive passenger flow, stations need to improve their intelligence and automation degree by an appropriate integrated system. Based on the existing integrated supervisory control system (ISCS) and simulation software (Anylogic), this paper designs an intelligent passenger organization system (IPOS) for metro stations. Its primary function includes passenger information acquisition, data processing and computing, visualization management, decision recommendations, and decision response based on interlocking equipment. For this purpose, the logical structure and intelligent algorithms employed are particularly devised. Besides, the structure diagram of information acquisition and application module, the application of Anylogic, the case library's function process are all given by this research. Based on the secondary development of Anylogic and existing technologies like video recognition, the IPOS is supposed to improve the response speed and address capacity in the face of emergent passenger flow of metro stations.

Keywords: anylogic software, decision-making support system, intellectualization, ISCS, passenger organization

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5738 Microstructure Dependent Fatigue Crack Growth in Aluminum Alloy

Authors: M. S. Nandana, K. Udaya Bhat, C. M. Manjunatha

Abstract:

In this study aluminum alloy 7010 was subjected to three different ageing treatments i.e., peak ageing (T6), over-ageing (T7451) and retrogression and re ageing (RRA) to study the influence of precipitate microstructure on the fatigue crack growth rate behavior. The microstructural modification was studied by using transmission electron microscope (TEM) to examine the change in the size and morphology of precipitates in the matrix and on the grain boundaries. The standard compact tension (CT) specimens were fabricated and tested under constant amplitude fatigue crack growth tests to evaluate the influence of heat treatment on the fatigue crack growth rate properties. The tests were performed in a computer-controlled servo-hydraulic test machine applying a load ratio, R = 0.1 at a loading frequency of 10 Hz as per ASTM E647. The fatigue crack growth was measured by adopting compliance technique using a CMOD gauge attached to the CT specimen. The average size of the matrix precipitates were found to be of 16-20 nm in T7451, 5-6 nm in RRA and 2-3 nm in T6 conditions respectively. The grain boundary precipitate which was continuous in T6, was disintegrated in RRA and T7451 condition. The PFZ width was lower in RRA compared to T7451 condition. The crack growth rate was higher in T7451 and lowest in RRA treated alloy. The RRA treated alloy also exhibits an increase in threshold stress intensity factor range (∆Kₜₕ). The ∆Kₜₕ measured was 11.1, 10.3 and 5.7 MPam¹/² in RRA, T6 and T7451 alloys respectively. The fatigue crack growth rate in RRA treated alloy was nearly 2-3 times lower than that in T6 and was one order lower than that observed in T7451 condition. The surface roughness of RRA treated alloy was more pronounced when compared to the other conditions. The reduction in fatigue crack growth rate in RRA alloy was majorly due to the increase in roughness and partially due to increase in spacing between the matrix precipitates. The reduction in crack growth rate and increase in threshold stress intensity range is expected to benefit the damage tolerant capability of aircraft structural components under service loads.

Keywords: damage tolerance, fatigue, heat treatment, PFZ, RRA

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5737 Memory-Guided Oculomotor Task in High School Football Players with ADHD, Post-Concussive Injuries, and Controls

Authors: B. McGovern, J. F. Luck, A. Gade, I. V. Lake, D. O’Connell, H. C. Cutcliffe, K. P. Shah, E. E. Ginalis, C. M. Lambert, N. Christian, J. R. Kait, A. W. Yu, C. P. Eckersley, C. R. Bass

Abstract:

Mild traumatic brain injury (mTBI) in the form of post-concussive injuries and attention deficit / hyperactivity disorder (ADHD) share similar cognitive impairments, including impaired working memory and executive function. The memory-guided oculomotor task separates working memory and inhibitory components to provide further information on the nature of these deficits in each pathology. Eleven subjects with ADHD, fifteen control subjects, and ten subjects with recent concussive injury were matched on age, gender, and education (all high school-age males). Eye movements were recorded during memory-guided oculomotor tasks with varying delays using EyeLink 1000 (SR Research). The percentage of premature saccades and the latency of correct response are the analyzed measures for response inhibition and working memory, respectively. No significant differences were found in latencies between controls subjects and subjects with ADHD or post-concussive injuries, in accordance with previous studies. Subjects with ADHD and post-concussive injuries both demonstrated a trend of increased percentages of premature saccades compared to control subjects in the same oculomotor task. This trend reached statistical significance between the post-concussive and control groups (p < 0.05). These findings support the primary nature of the executive function deficits in response inhibition in ADHD and mTBI. The interpretation of results is limited by the small sample size and the exploratory nature of the study. Further investigation into oculomotor performance differences in mTBI and ADHD may help in differentiating these pathologies in consequent diagnoses and provide insight into the interaction of these deficits in mTBI.

Keywords: attention deficit / hyperactivity disorder (ADHD), concussion, diagnosis, oculomotor, pediatrics

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5736 Investigation of Bird Impact on Tailplane

Authors: Reza Hedayati, Meysam Jahanbakhshi

Abstract:

The typical airplane stabilizer structures consist of two main similar segments (outer and inner parts), one of them a little larger than the other. In this study, bird impact on four different spots of the stabilizer structure: (a) between two ribs of smaller segment, (b) between two ribs of larger segment, (c) on the rib connecting the two segments, and (d) on a middle rib of the smaller segment, is investigated and their results are compared by means of energy absorption, displacement, and bird’s mass diagrams as well as visible damage induced on the stabilizer structure.

Keywords: airplane, bird strike, LS-DYNA, stabilizer

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5735 A Lesson in the Social Welfare System in Mexico: Limited Resources for Unlimited Needs

Authors: Vanessa L. Haro

Abstract:

Beginning with a historical foundation of Mexico, this marks the start of a close examination of this major Latin American country by providing the context needed to understand the reasons for Mexico’s strengths and struggles today, specific to their response to the issue of gender violence. Responding to the challenge of combating gender violence and inequality, Mexico has created social programs and initiatives in hopes of addressing these issues and modernizing their gender norms, which currently disempower and dehumanize women, while simultaneously denying women the necessary tools needed to fight back or bring balance to the gender scales. Nevertheless, women in Mexico have made their voices heard with the most salient image of that of the mothers protesting while holding the photos of their young daughters who lost their lives. This case study on gender issues in Mexico works to acknowledge the diverse forces that contribute to the issue of gender violence, and to make a statement that this is a crisis that requires a more dynamic response within Mexico’s social welfare policies, and should not be allowed to continue to progress as a normative phenomenon. As the advocacy groups and protesters cry out, “Ni una menos! (Not one less), meaning we will not lose one more woman and making the statement that all women’s lives matter.

Keywords: gender issues, Mexico, poverty, social welfare

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5734 Optimization in the Compressive Strength of Iron Slag Self-Compacting Concrete

Authors: Luis E. Zapata, Sergio Ruiz, María F. Mantilla, Jhon A. Villamizar

Abstract:

Sand as fine aggregate for concrete production needs a feasible substitute due to several environmental issues. In this work, a study of the behavior of self-compacting concrete mixtures under replacement of sand by iron slag from 0.0% to 50.0% of weight and variations of water/cementitious material ratio between 0.3 and 0.5 is presented. Control fresh state tests of Slump flow, T500, J-ring and L-box were determined. In the hardened state, compressive strength was determined and optimization from response surface analysis was performed. The study of the variables in the hardened state was developed based on inferential statistical analyses using central composite design methodology and posterior analyses of variance (ANOVA). An increase in the compressive strength up to 50% higher than control mixtures at 7, 14, and 28 days of maturity was the most relevant result regarding the presence of iron slag as replacement of natural sand. Considering the obtained result, it is possible to infer that iron slag is an acceptable alternative replacement material of the natural fine aggregate to be used in structural concrete.

Keywords: ANOVA, iron slag, response surface analysis, self-compacting concrete

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5733 Involvement of Nrf2 in Kolaviron-Mediated Attenuation of Behavioural Incompetence and Neurodegeneration in a Murine Model of Parkinson's Disease

Authors: Yusuf E. Mustapha, Inioluwa A Akindoyeni, Oluwatoyin G. Ezekiel, Ifeoluwa O. Awogbindin, Ebenezer O. Farombi

Abstract:

Background: Parkinson's disease (PD) is the most prevalent motor disorder. Available therapies are palliative with no effect on disease progression. Kolaviron (KV), a natural anti-inflammatory and antioxidant agent, has been reported to possess neuroprotective effects in Parkinsonian flies and rats. Objective: The present study investigates the neuroprotective effect of KV, focusing on the DJ1/Nrf2 signaling pathway. Methodology: All-trans retinoic acid (ATRA, 10 mg/kg, i.p.) was used to inhibit Nrf2. Murine model of PD was established with four doses of MPTP (20 mg/kg i.p.) at 2 hours interval. MPTP mice were pre-treated with either KV (200 mg/kg/day p.o), ATRA, or both conditions for seven days before PD induction. Motor behaviour was evaluated, and markers of oxidative stress/damage and its regulators were assessed with immunofluorescence and ELISA techniques. Results: MPTP-treated mice covered less distance with reduced numbers of anticlockwise rotations, heightened freezing, and prolonged immobility when compared to control. However, KV significantly attenuated these deficits. Pretreatment of MPTP mice with KV upregulated Nrf2 expression beyond MPTP level with a remarkable reduction in Keap1 expression and marked elevation of DJ-1 level, whereas co-administration with ATRA abrogated these effects. KV treatment restored MPTP-mediated depletion of endogenous antioxidant, striatal oxidative stress, oxidative damage, and inhibition of acetylcholinesterase activity. However, ATRA treatment potentiated acetylcholinesterase inhibition and attenuated the protective effect of KV on the level of nitric oxide and activities of catalase and superoxide dismutase. Conclusion: Kolaviron protects Parkinsonian mice by stabilizing and activating the Nrf2 signaling pathway. Thus, kolaviron can be explored as a pharmacological lead in PD management.

Keywords: Garcinia kola, Kolaviron, Parkinson Disease, Nrf2, behavioral incompetence, neurodegeneration

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5732 Multi-Criteria Decision-Making Evaluations for Oily Waste Management of Marine Oil Spill

Authors: Naznin Sultana Daisy, Mohammad Hesam Hafezi, Lei Liu

Abstract:

Nowadays, oily solid waste management has become an important issue for many countries due to frequent oil spill accidents and the increase of industrial oily wastewater. The historical oil spill data show that marine oil spills that affect the shoreline can, in extreme cases, produce up to 30 or 40 times more waste than the volume of oil initially released. Hence, responsive authorities aim to develop the most effective oily waste management solution in a timely manner to manage and minimize the waste generated. In this study initially, we tried to develop the roadmap of oily waste management for three-tiered spill scenarios for Atlantic Canada. For that purpose, three oily waste disposal scenarios are evaluated via six criteria which are determined according to the opinions of the experts from the field. Consequently, through sustainable response strategies, the most appropriate and feasible scenario is determined. The results of this study will assist to develop an integrated oily waste management system for identifying the optimal waste-generation-allocation-disposal schemes and generating the optimal management alternatives based on the holistic consideration of environmental, technological, economic, social, and regulatory factors.

Keywords: oily waste management, marine oil spill, multi-criteria decision making, oil spill response

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5731 A Review of Pharmacological Prevention of Peri-and Post-Procedural Myocardial Injury After Percutaneous Coronary Intervention

Authors: Syed Dawood Md. Taimur, Md. Hasanur Rahman, Syeda Fahmida Afrin, Farzana Islam

Abstract:

The concept of myocardial injury, although first recognized from animal studies, is now recognized as a clinical phenomenon that may result in microvascular damage, no-reflow phenomenon, myocardial stunning, myocardial hibernation and ischemic preconditioning. The final consequence of this event is left ventricular (LV) systolic dysfunction leading to increased morbidity and mortality. The typical clinical case of reperfusion injury occurs in acute myocardial infarction (MI) with ST segment elevation in which an occlusion of a major epicardial coronary artery is followed by recanalization of the artery. This may occur either spontaneously or by means of thrombolysis and/or by primary percutaneous coronary intervention (PCI) with efficient platelet inhibition by aspirin (acetylsalicylic acid), clopidogrel and glycoprotein IIb/IIIa inhibitors. In recent years, percutaneous coronary intervention (PCI) has become a well-established technique for the treatment of coronary artery disease. PCI improves symptoms in patients with coronary artery disease and it has been increasing the safety of procedures. However, peri- and post-procedural myocardial injury, including angiographical slow coronary flow, microvascular embolization, and elevated levels of cardiac enzyme, such as creatine kinase and troponin-T and -I, has also been reported even in elective cases. Furthermore, myocardial reperfusion injury at the beginning of myocardial reperfusion, which causes tissue damage and cardiac dysfunction, may occur in cases of the acute coronary syndrome. Because patients with myocardial injury is related to larger myocardial infarction and have a worse long-term prognosis than those without myocardial injury, it is important to prevent myocardial injury during and/or after PCI in patients with coronary artery disease. To date, many studies have demonstrated that adjunctive pharmacological treatment suppresses myocardial injury and increases coronary blood flow during PCI procedures. In this review, we highlight the usefulness of pharmacological treatment in combination with PCI in attenuating myocardial injury in patients with coronary artery disease.

Keywords: coronary artery disease, percutaneous coronary intervention, myocardial injury, pharmacology

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5730 The X-Ray Response Team: Building a National Health Pre-Hospital Service

Authors: Julian Donovan, Jessica Brealey, Matthew Bowker, Marianne Feghali, Gregory Smith, Lee Thompson, Deborah Henderson

Abstract:

This article details the development of the X-ray response team (XRT), a service that utilises innovative technology to safely deliver acute and elective imaging and medical assessment service in the pre-hospital and community setting. This involves a partnership between Northumbria Healthcare NHS Foundation Trust’s Radiology and Emergency Medicine departments and the North East Ambulance Service to create a multidisciplinary prehospital team. The team committed to the delivery of a two-day acute service every week, alongside elective referrals, starting in November 2020. The service was originally made available to a 15-mile radius surrounding the Northumbria Hospital. Due to demand, this was expanded to include the North Tyneside and Northumberland regions. The target population was specified as frail and vulnerable patients, as well as those deemed to benefit from staying in their own environment. Within the first two months, thirty-six percent of patients assessed were able to stay at home due to the provision of off-site imaging. In the future, this service aims to allow patient transfer directly to an appropriate ward or clinic, bypassing the emergency department to improve the patient journey and reduce emergency care pressures.

Keywords: frailty, imaging, pre-hospital, X-ray

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5729 Influence of Pile Radius on Inertial Response of Pile Group in Fundamental Frequency of Homogeneous Soil Medium

Authors: Faghihnia Torshizi Mostafa, Saitoh Masato

Abstract:

An efficient method is developed for the response of a group of vertical, cylindrical fixed-head, finite length piles embedded in a homogeneous elastic stratum, subjected to harmonic force atop the pile group cap. Pile to pile interaction is represented through simplified beam-on-dynamic-Winkler-foundation (BDWF) with realistic frequency-dependent springs and dashpots. Pile group effect is considered through interaction factors. New closed-form expressions for interaction factors and curvature ratios atop the pile are extended by considering different boundary conditions at the tip of the piles (fixed, hinged). In order to investigate the fundamental characteristics of inertial bending strains in pile groups, inertial bending strains at the head of each pile are expressed in terms of slenderness ratio. The results of parametric study give valuable insight in understanding the behavior of fixed head pile groups in fundamental natural frequency of soil stratum.

Keywords: Winkler-foundation, fundamental frequency of soil stratum, normalized inertial bending strain, harmonic excitation

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5728 Research on the Environmental Assessment Index of Brownfield Redevelopment in Taiwan: A Case Study on Formosa Chemicals and Fibre Corporation, Changhua Branch

Authors: Min-Chih Yang, Shih-Jen Feng, Bo-Tsang Li

Abstract:

The concept of “Brownfield” has been developed for nearly 35 years since it was put forward in 《Comprehensive Environmental Response, Compensation, and Liability Act, CERCLA》of USA in 1980 for solving the problem of soil contamination of those old industrial lands, and later, many countries have put forward relevant policies and researches continuously. But the related concept in Taiwan, a country has developed its industry for 60 years, is still in its infancy. This leads to the slow development of Brownfield related research and policy in Taiwan. When it comes to build the foundation of Brownfield development, we have to depend on the related experience and research of other countries. They are four aspects about Brownfield: 1. Contaminated Land; 2. Derelict Land; 3. Vacant Land; 4. Previously Development Land. This study will focus on and deeply investigate the Vacant land and contaminated land. The subject of this study is Formosa Chemicals & Fibre Corporation, Changhua branch in Taiwan. It has been operating for nearly 50 years and contributing a lot to the local economy. But under the influence of the toxic waste and sewage which was drained regularly or occasionally out from the factory, the environment has been destroyed seriously. There are three factors of pollution: 1. environmental toxicants, carbon disulfide, released from producing processes and volatile gases which is hard to monitor; 2. Waste and exhaust gas leakage caused by outdated equipment; 3. the wastewater discharge has seriously damage the ecological environment of the Dadu river estuary. Because of all these bad influences, the factory has been closed nowadays and moved to other places to spare the opportunities for the contaminated lands to re-develop. So we collect information about related Brownfield management experience and policies in different countries as background information to investigate the current Taiwanese Brownfield redevelopment issues and built the environmental assessment framework for it. We hope that we can set the environmental assessment indexes for Formosa Chemicals & Fibre Corporation, Changhua branch according to the framework. By investigating the theory and environmental pollution factors, we will carry out deep analysis and expert questionnaire to set those indexes and prove a sample in Taiwan for Brownfield redevelopment and remediation in the future.

Keywords: brownfield, industrial land, redevelopment, assessment index

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5727 Formative Assessment in an Introductory Python Programming Course

Authors: María José Núñez-Ruiz, Luis Álvarez-González, Cristian Olivares-Rodriguez, Benjamin Lazo-Letelier

Abstract:

This paper begins with some concept of formative assessment and the relationship with learning objective: contents objectives, processes objectives, and metacognitive objectives. Two methodologies are describes Evidence-Based teaching and Question Drive Instruction. To do formative assessments in larges classes a Classroom Response System (CRS) is needed. But most of CRS use only Multiple Choice Questions (MCQ), True/False question, or text entry; however, this is insufficient to formative assessment. To do that a new CRS, call FAMA was developed. FAMA support six types of questions: Choice, Order, Inline choice, Text entry, Associated, and Slider. An experiment participated in 149 students from four engineering careers. For results, Kendall's Range Correlation Analysis and descriptive analysis was done. In conclusion, there is a strong relation between contents question, process questions (ask in formative assessment without a score) and metacognitive questions, asked in summative assessment. As future work, the lecturer can do personalized teaching, because knows the behavior of all students in each formative assessment

Keywords: Python language, formative assessment, classroom response systems, evidence-Based teaching, question drive instruction

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5726 Aspects Regarding the Structural Behaviour of Autonomous Underwater Vehicle for Emergency Response

Authors: Lucian Stefanita Grigore, Damian Gorgoteanu, Cristian Molder, Amado Stefan, Daniel Constantin

Abstract:

The purpose of this article is to present an analytical-numerical study on the structural behavior of a sunken autonomous underwater vehicle (AUV) for emergency intervention. The need for such a study was generated by the key objective of the ERL-Emergency project. The project aims to develop a system of collaborative robots for emergency response. The system consists of two robots: unmanned ground vehicles (UGV) on tracks and the second is an AUV. The system of collaborative robots, AUV and UGV, will be used to perform missions of monitoring, intervention, and rescue. The main mission of the AUV is to dive into the maritime space of an industrial port to detect possible leaks in a pipeline transporting petroleum products. Another mission is to close and open the valves with which the pipes are provided. Finally, you will need to be able to lift a manikin to the surface, which you can take to land. Numerical analysis was performed by the finite element method (FEM). The conditions for immersing the AUV at 100 m depth were simulated, and the calculations for different fluid flow rates were repeated. From a structural point of view, the stiffening areas and the enclosures in which the command-and-control elements and the accumulators are located have been especially analyzed. The conclusion of this research is that the AUV meets very well the established requirements.

Keywords: analytical-numerical, emergency, FEM, robotics, underwater

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5725 Treatment of Non-Small Cell Lung Cancer (NSCLC) With Activating Mutations Considering ctDNA Fluctuations

Authors: Moiseenko F. V., Volkov N. M., Zhabina A. S., Stepanova E. O., Kirillov A. V., Myslik A. V., Artemieva E. V., Agranov I. R., Oganesyan A. P., Egorenkov V. V., Abduloeva N. H., Aleksakhina S. Yu., Ivantsov A. O., Kuligina E. S., Imyanitov E. N., Moiseyenko V. M.

Abstract:

Analysis of ctDNA in patients with NSCLC is an emerging biomarker. Multiple research efforts of quantitative or at least qualitative analysis before and during the first periods of treatment with TKI showed the prognostic value of ctDNA clearance. Still, these important results are not incorporated in clinical standards. We evaluated the role of ctDNA in EGFR-mutated NSCLC receiving first-line TKI. Firstly, we analyzed sequential plasma samples from 30 patients that were collected before intake of the first tablet (at baseline) and at 6, 12, 24, 36, and 48 hours after the “starting point.” EGFR-M+ allele was measured by ddPCR. Afterward, we included sequential qualitative analysis of ctDNA with cobas® EGFR Mutation Test v2 from 99 NSCLC patients before the first dose, after 2 and 4 months of treatment, and on progression. Early response analysis showed the decline of EGFR-M+ level in plasma within the first 48 hours of treatment in 11 subjects. All these patients showed objective tumor response. 10 patients showed either elevation of EGFR-M+ plasma concentration (n = 5) or stable content of circulating EGFR-M+ after the start of the therapy (n = 5); only 3 of these patients achieved an objective response (p = 0.026) when compared to the former group). The rapid decline of plasma EGFR-M+ DNA concentration also predicted for longer PFS (13.7 vs. 11.4 months, p = 0.030). Long-term ctDNA monitoring showed clinically significant heterogeneity of EGFR-mutated NSCLC treated with 1st line TKIs in terms of progression-free and overall survival. Patients without detectable ctDNA at baseline (N = 32) possess the best prognosis on the duration of treatment (PFS: 24.07 [16.8-31.3] and OS: 56.2 [21.8-90.7] months). Those who achieve clearance after two months of TKI (N = 42) have indistinguishably good PFS (19.0 [13.7 – 24.2]). Individuals who retain ctDNA after 2 months (N = 25) have the worst prognosis (PFS: 10.3 [7.0 – 13.5], p = 0.000). 9/25 patients did not develop ctDNA clearance at 4 months with no statistical difference in PFS from those without clearance at 2 months. Prognostic heterogeneity of EGFR-mutated NSCLC should be taken into consideration in planning further clinical trials and optimizing the outcomes of patients.

Keywords: NSCLC, EGFR, targeted therapy, ctDNA, prognosis

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5724 Self-Assembled Laser-Activated Plasmonic Substrates for High-Throughput, High-Efficiency Intracellular Delivery

Authors: Marinna Madrid, Nabiha Saklayen, Marinus Huber, Nicolas Vogel, Christos Boutopoulos, Michel Meunier, Eric Mazur

Abstract:

Delivering material into cells is important for a diverse range of biological applications, including gene therapy, cellular engineering and imaging. We present a plasmonic substrate for delivering membrane-impermeable material into cells at high throughput and high efficiency while maintaining cell viability. The substrate fabrication is based on an affordable and fast colloidal self-assembly process. When illuminated with a femtosecond laser, the light interacts with the electrons at the surface of the metal substrate, creating localized surface plasmons that form bubbles via energy dissipation in the surrounding medium. These bubbles come into close contact with the cell membrane to form transient pores and enable entry of membrane-impermeable material via diffusion. We use fluorescence microscopy and flow cytometry to verify delivery of membrane-impermeable material into HeLa CCL-2 cells. We show delivery efficiency and cell viability data for a range of membrane-impermeable cargo, including dyes and biologically relevant material such as siRNA. We estimate the effective pore size by determining delivery efficiency for hard fluorescent spheres with diameters ranging from 20 nm to 2 um. To provide insight to the cell poration mechanism, we relate the poration data to pump-probe measurements of micro- and nano-bubble formation on the plasmonic substrate. Finally, we investigate substrate stability and reusability by using scanning electron microscopy (SEM) to inspect for damage on the substrate after laser treatment. SEM images show no visible damage. Our findings indicate that self-assembled plasmonic substrates are an affordable tool for high-throughput, high-efficiency delivery of material into mammalian cells.

Keywords: femtosecond laser, intracellular delivery, plasmonic, self-assembly

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5723 Physical and Chemical Alternative Methods of Fresh Produce Disinfection

Authors: Tuji Jemal Ahmed

Abstract:

Fresh produce is an essential component of a healthy diet. However, it can also be a potential source of pathogenic microorganisms that can cause foodborne illnesses. Traditional disinfection methods, such as washing with water and chlorine, have limitations and may not effectively remove or inactivate all microorganisms. This has led to the development of alternative/new methods of fresh produce disinfection, including physical and chemical methods. In this paper, we explore the physical and chemical new methods of fresh produce disinfection, their advantages and disadvantages, and their suitability for different types of produce. Physical methods of disinfection, such as ultraviolet (UV) radiation and high-pressure processing (HPP), are crucial in ensuring the microbiological safety of fresh produce. UV radiation uses short-wavelength UV-C light to damage the DNA and RNA of microorganisms, and HPP applies high levels of pressure to fresh produce to reduce the microbial load. These physical methods are highly effective in killing a wide range of microorganisms, including bacteria, viruses, and fungi. However, they may not penetrate deep enough into the product to kill all microorganisms and can alter the sensory characteristics of the product. Chemical methods of disinfection, such as acidic electrolyzed water (AEW), ozone, and peroxyacetic acid (PAA), are also important in ensuring the microbiological safety of fresh produce. AEW uses a low concentration of hypochlorous acid and a high concentration of hydrogen ions to inactivate microorganisms, ozone uses ozone gas to damage the cell membranes and DNA of microorganisms, and PAA uses a combination of hydrogen peroxide and acetic acid to inactivate microorganisms. These chemical methods are highly effective in killing a wide range of microorganisms, but they may cause discoloration or changes in the texture and flavor of some products and may require specialized equipment and trained personnel to produce and apply. In conclusion, the selection of the most suitable method of fresh produce disinfection should take into consideration the type of product, the level of microbial contamination, the effectiveness of the method in reducing the microbial load, and any potential negative impacts on the sensory characteristics, nutritional composition, and safety of the produce.

Keywords: fresh produce, pathogenic microorganisms, foodborne illnesses, disinfection methods

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5722 Nafion Multiwalled Carbon Nano Tubes Composite Film Modified Glassy Carbon Sensor for the Voltammetric Estimation of Dianabol Steroid in Pharmaceuticals and Biological Fluids

Authors: Nouf M. Al-Ourfi, A. S. Bashammakh, M. S. El-Shahawi

Abstract:

The redox behavior of dianabol steroid (DS) on Nafion Multiwalled Carbon nano -tubes (MWCNT) composite film modified glassy carbon electrode (GCE) in various buffer solutions was studied using cyclic voltammetry (CV) and differential pulse- adsorptive cathodic stripping voltammetry (DP-CSV) and successfully compared with the results at non modified bare GCE. The Nafion-MWCNT composite film modified GCE exhibited the best electrochemical response among the two electrodes for the electro reduction of DS that was inferred from the EIS, CV and DP-CSV. The modified sensor showed a sensitive, stable and linear response in the concentration range of 5 – 100 nM with a detection limit of 0.08 nM. The selectivity of the proposed sensor was assessed in the presence of high concentration of major interfering species. The analytical application of the sensor for the quantification of DS in pharmaceutical formulations and biological fluids (urine) was determined and the results demonstrated acceptable recovery and RSD of 5%. Statistical treatment of the results of the proposed method revealed no significant differences in the accuracy and precision. The relative standard deviations for five measurements of 50 and 300 ng mL−1 of DS were 3.9 % and 1.0 %, respectively.

Keywords: dianabol steroid, determination, modified GCE, urine

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5721 Distance and Coverage: An Assessment of Location-Allocation Models for Fire Stations in Kuwait City, Kuwait

Authors: Saad M. Algharib

Abstract:

The major concern of planners when placing fire stations is finding their optimal locations such that the fire companies can reach fire locations within reasonable response time or distance. Planners are also concerned with the numbers of fire stations that are needed to cover all service areas and the fires, as demands, with standard response time or distance. One of the tools for such analysis is location-allocation models. Location-allocation models enable planners to determine the optimal locations of facilities in an area in order to serve regional demands in the most efficient way. The purpose of this study is to examine the geographic distribution of the existing fire stations in Kuwait City. This study utilized location-allocation models within the Geographic Information System (GIS) environment and a number of statistical functions to assess the current locations of fire stations in Kuwait City. Further, this study investigated how well all service areas are covered and how many and where additional fire stations are needed. Four different location-allocation models were compared to find which models cover more demands than the others, given the same number of fire stations. This study tests many ways to combine variables instead of using one variable at a time when applying these models in order to create a new measurement that influences the optimal locations for locating fire stations. This study also tests how location-allocation models are sensitive to different levels of spatial dependency. The results indicate that there are some districts in Kuwait City that are not covered by the existing fire stations. These uncovered districts are clustered together. This study also identifies where to locate the new fire stations. This study provides users of these models a new variable that can assist them to select the best locations for fire stations. The results include information about how the location-allocation models behave in response to different levels of spatial dependency of demands. The results show that these models perform better with clustered demands. From the additional analysis carried out in this study, it can be concluded that these models applied differently at different spatial patterns.

Keywords: geographic information science, GIS, location-allocation models, geography

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5720 4D Monitoring of Subsurface Conditions in Concrete Infrastructure Prior to Failure Using Ground Penetrating Radar

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

Abstract:

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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5719 Response of Pavement under Temperature and Vehicle Coupled Loading

Authors: Yang Zhong, Mei-Jie Xu

Abstract:

To study the dynamic mechanics response of asphalt pavement under the temperature load and vehicle loading, asphalt pavement was regarded as multilayered elastic half-space system, and theory analysis was conducted by regarding dynamic modulus of asphalt mixture as the parameter. Firstly, based on the dynamic modulus test of asphalt mixture, function relationship between the dynamic modulus of representative asphalt mixture and temperature was obtained. In addition, the analytical solution for thermal stress in the single layer was derived by using Laplace integral transformation and Hankel integral transformation respectively by using thermal equations of equilibrium. The analytical solution of calculation model of thermal stress in asphalt pavement was derived by transfer matrix of thermal stress in multilayer elastic system. Finally, the variation of thermal stress in pavement structure was analyzed. The result shows that there is an obvious difference between the thermal stress based on dynamic modulus and the solution based on static modulus. Therefore, the dynamic change of parameter in asphalt mixture should be taken into consideration when the theoretical analysis is taken out.

Keywords: asphalt pavement, dynamic modulus, integral transformation, transfer matrix, thermal stress

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5718 Observation of Critical Sliding Velocity

Authors: Visar Baxhuku, Halil Demolli, Alishukri Shkodra

Abstract:

This paper presents the monitoring of vehicle movement, namely the developing of speed of vehicles during movement in a certain twist. The basic geometry data of twist are measured with the purpose of calculating the slide in border speed. During the research, measuring developed speed of passenger vehicles for the real conditions of the road surface, dry road with average damage, was realised. After setting values, the analysis was done in function security of movement in twist.

Keywords: critical sliding velocity, moving velocity, curve, passenger vehicles

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5717 Ubuntu: A Holistic Social Framework for Preserving Ecosystem Amidst the Climate Change Challenges

Authors: Gabriel Sunday Ayayia

Abstract:

The paper argues that Ubuntu, as a philosophy that emphasizes the interconnectedness of all living things and importance of community and mutual support, can be used as a social framework to address the problems of climate change and promote environmental sustainability. The research demonstrate that Ubuntu is an ideological concept that encourages collective action on climate change, with the emphasis on individual and collective commitment to taking concrete action to address the problems of climate change. The paper shows that Ubuntu can be employed as a social tool that would enhance the cultivation of shared identity and promote the sense of shared response responsibility to develop the resilience to cope with climate change. Using qualitative and quantitative methodologies, the study establishes the imperativeness of mutual support and cooperation through the lens of Ubuntu as a human-centered scalable response to the debacle of climate change. It recommends that we can build a society that values the environment and promotes sustainable practices by encouraging community involvement in sustainable initiatives by integrating Ubuntu-based principles to our decision-making processes, collaboration, leadership, human agency and governance.

Keywords: ubuntu, climate change, humanity, collective actions, community-based

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5716 Case Study Hyperbaric Oxygen Therapy for Idiopathic Sudden Sensorineural Hearing Loss

Authors: Magdy I. A. Alshourbagi

Abstract:

Background: The National Institute for Deafness and Communication Disorders defines idiopathic sudden sensorineural hearing loss as the idiopathic loss of hearing of at least 30 dB across 3 contiguous frequencies occurring within 3 days.The most common clinical presentation involves an individual experiencing a sudden unilateral hearing loss, tinnitus, a sensation of aural fullness and vertigo. The etiologies and pathologies of ISSNHL remain unclear. Several pathophysiological mechanisms have been described including: vascular occlusion, viral infections, labyrinthine membrane breaks, immune associated disease, abnormal cochlear stress response, trauma, abnormal tissue growth, toxins, ototoxic drugs and cochlear membrane damage. The rationale for the use of hyperbaric oxygen to treat ISSHL is supported by an understanding of the high metabolism and paucity of vascularity to the cochlea. The cochlea and the structures within it require a high oxygen supply. The direct vascular supply, particularly to the organ of Corti, is minimal. Tissue oxygenation to the structures within the cochlea occurs via oxygen diffusion from cochlear capillary networks into the perilymph and the cortilymph. . The perilymph is the primary oxygen source for these intracochlear structures. Unfortunately, perilymph oxygen tension is decreased significantly in patients with ISSHL. To achieve a consistent rise of perilymph oxygen content, the arterial-perilymphatic oxygen concentration difference must be extremely high. This can be restored with hyperbaric oxygen therapy. Subject and Methods: A 37 year old man was presented at the clinic with a five days history of muffled hearing and tinnitus of the right ear. Symptoms were sudden onset, with no associated pain, dizziness or otorrhea and no past history of hearing problems or medical illness. Family history was negative. Physical examination was normal. Otologic examination revealed normal tympanic membranes bilaterally, with no evidence of cerumen or middle ear effusion. Tuning fork examination showed positive Rinne test bilaterally but with lateralization of Weber test to the left side, indicating right ear sensorineural hearing loss. Audiometric analysis confirmed sensorineural hearing loss across all frequencies of about 70- dB in the right ear. Routine lab work were all within normal limits. Clinical diagnosis of idiopathic sudden sensorineural hearing loss of the right ear was made and the patient began a medical treatment (corticosteroid, vasodilator and HBO therapy). The recommended treatment profile consists of 100% O2 at 2.5 atmospheres absolute for 60 minutes daily (six days per week) for 40 treatments .The optimal number of HBOT treatments will vary, depending on the severity and duration of symptomatology and the response to treatment. Results: As HBOT is not yet a standard for idiopathic sudden sensorineural hearing loss, it was introduced to this patient as an adjuvant therapy. The HBOT program was scheduled for 40 sessions, we used a 12-seat multi place chamber for the HBOT, which was started at day seven after the hearing loss onset. After the tenth session of HBOT, improvement of both hearing (by audiogram) and tinnitus was obtained in the affected ear (right). Conclusions: In conclusion, HBOT may be used for idiopathic sudden sensorineural hearing loss as an adjuvant therapy. It may promote oxygenation to the inner ear apparatus and revive hearing ability. Patients who fail to respond to oral and intratympanic steroids may benefit from this treatment. Further investigation is warranted, including animal studies to understand the molecular and histopathological aspects of HBOT and randomized control clinical studies.

Keywords: idiopathic sudden sensorineural hearing loss (issnhl), hyperbaric oxygen therapy (hbot), the decibel (db), oxygen (o2)

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5715 Healthy Beverages Made from Grape Juice: Antioxidant, Energetic, and Isotonic Components

Authors: Yasmina Bendaali, Cristian Vaquero, Carlos Escott, Carmen González, Antonio Morata

Abstract:

Consumer tendencies to healthy eating habits and request for organic beverages led to the production of new drinks from fruit juices as a source of nutrients and bioactive compounds. Grape juice is a rich source of sugars, organic acids, and phenolic compounds, which define its beneficial effect on health and the attractive sensory profile for consumers' choices (color, taste, flavor). Thus, grape juice was used as a source of sugars, avoiding the addition of sweeteners by diluting it with mineral water to obtain the sugar concentration recommended for isotonic drinks (6% to 8%) to provide energy during physical activities. In addition, phenolic compounds of grape juice are associated with many human health benefits, mainly antioxidant activity, which helps to prevent different diseases associated with oxidative stress, including cancers and cardiovascular and neurodegenerative diseases. Furthermore, physical exercise has been shown to increase the production of free radicals and other reactive oxygen species. Thus, athletes need to improve their antioxidant defense systems to prevent oxidative damage. Different studies have demonstrated the positive effect of grape juice consumption during physical activities, which improves antioxidant activity and performance, protects against oxidative damage, and reduces inflammation. Thus, the use of grape juice to develop isotonic drinks can provide isotonic drinks with antioxidant and biological activities in addition to their principal role of rehydration and replacement of minerals and carbohydrates during physical exercises. Moreover, attractive sensory characteristics, mainly color, which is provided by anthocyanin content, have a great contribution to making the drinks more natural and help to dispense the use of synthetic dyes in addition to the health benefits which will be a novel product in the field of healthy beverages responding on the demand of consumers for new, innovative, and healthy products.

Keywords: grape juice, isotonic, antioxidants, anthocyanins, natural, sport

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5714 Validation and Fit of a Biomechanical Bipedal Walking Model for Simulation of Loads Induced by Pedestrians on Footbridges

Authors: Dianelys Vega, Carlos Magluta, Ney Roitman

Abstract:

The simulation of loads induced by walking people in civil engineering structures is still challenging It has been the focus of considerable research worldwide in the recent decades due to increasing number of reported vibration problems in pedestrian structures. One of the most important key in the designing of slender structures is the Human-Structure Interaction (HSI). How moving people interact with structures and the effect it has on their dynamic responses is still not well understood. To rely on calibrated pedestrian models that accurately estimate the structural response becomes extremely important. However, because of the complexity of the pedestrian mechanisms, there are still some gaps in knowledge and more reliable models need to be investigated. On this topic several authors have proposed biodynamic models to represent the pedestrian, whether these models provide a consistent approximation to physical reality still needs to be studied. Therefore, this work comes to contribute to a better understanding of this phenomenon bringing an experimental validation of a pedestrian walking model and a Human-Structure Interaction model. In this study, a bi-dimensional bipedal walking model was used to represent the pedestrians along with an interaction model which was applied to a prototype footbridge. Numerical models were implemented in MATLAB. In parallel, experimental tests were conducted in the Structures Laboratory of COPPE (LabEst), at Federal University of Rio de Janeiro. Different test subjects were asked to walk at different walking speeds over instrumented force platforms to measure the walking force and an accelerometer was placed at the waist of each subject to measure the acceleration of the center of mass at the same time. By fitting the step force and the center of mass acceleration through successive numerical simulations, the model parameters are estimated. In addition, experimental data of a walking pedestrian on a flexible structure was used to validate the interaction model presented, through the comparison of the measured and simulated structural response at mid span. It was found that the pedestrian model was able to adequately reproduce the ground reaction force and the center of mass acceleration for normal and slow walking speeds, being less efficient for faster speeds. Numerical simulations showed that biomechanical parameters such as leg stiffness and damping affect the ground reaction force, and the higher the walking speed the greater the leg length of the model. Besides, the interaction model was also capable to estimate with good approximation the structural response, that remained in the same order of magnitude as the measured response. Some differences in frequency spectra were observed, which are presumed to be due to the perfectly periodic loading representation, neglecting intra-subject variabilities. In conclusion, this work showed that the bipedal walking model could be used to represent walking pedestrians since it was efficient to reproduce the center of mass movement and ground reaction forces produced by humans. Furthermore, although more experimental validations are required, the interaction model also seems to be a useful framework to estimate the dynamic response of structures under loads induced by walking pedestrians.

Keywords: biodynamic models, bipedal walking models, human induced loads, human structure interaction

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5713 A Comparison of qCON/qNOX to the Bispectral Index as Indices of Antinociception in Surgical Patients Undergoing General Anesthesia with Laryngeal Mask Airway

Authors: Roya Yumul, Ofelia Loani Elvir-Lazo, Sevan Komshian, Ruby Wang, Jun Tang

Abstract:

BACKGROUND: An objective means for monitoring the anti-nociceptive effects of perioperative medications has long been desired as a way to provide anesthesiologists information regarding a patient’s level of antinociception and preclude any untoward autonomic responses and reflexive muscular movements from painful stimuli intraoperatively. To this end, electroencephalogram (EEG) based tools including BIS and qCON were designed to provide information about the depth of sedation while qNOX was produced to inform on the degree of antinociception. The goal of this study was to compare the reliability of qCON/qNOX to BIS as specific indicators of response to nociceptive stimulation. METHODS: Sixty-two patients undergoing general anesthesia with LMA were included in this study. Institutional Review Board (IRB) approval was obtained, and informed consent was acquired prior to patient enrollment. Inclusion criteria included American Society of Anesthesiologists (ASA) class I-III, 18 to 80 years of age, and either gender. Exclusion criteria included the inability to consent. Withdrawal criteria included conversion to the endotracheal tube and EEG malfunction. BIS and qCON/qNOX electrodes were simultaneously placed on all patients prior to induction of anesthesia and were monitored throughout the case, along with other perioperative data, including patient response to noxious stimuli. All intraoperative decisions were made by the primary anesthesiologist without influence from qCON/qNOX. Student’s t-distribution, prediction probability (PK), and ANOVA were used to statistically compare the relative ability to detect nociceptive stimuli for each index. Twenty patients were included for the preliminary analysis. RESULTS: A comparison of overall intraoperative BIS, qCON and qNOX indices demonstrated no significant difference between the three measures (N=62, p> 0.05). Meanwhile, index values for qNOX (62±18) were significantly higher than those for BIS (46±14) and qCON (54±19) immediately preceding patient responses to nociceptive stimulation in a preliminary analysis (N=20, * p= 0.0408). Notably, certain hemodynamic measurements demonstrated a significant increase in response to painful stimuli (MAP increased from 74 ±13 mm Hg at baseline to 84 ± 18 mm Hg during noxious stimuli [p= 0.032] and HR from 76 ± 12 BPM at baseline to 80 ± 13 BPM during noxious stimuli [p=0.078] respectively). CONCLUSION: In this observational study, BIS and qCON/qNOX provided comparable information on patients’ level of sedation throughout the course of an anesthetic. Meanwhile, increases in qNOX values demonstrated a superior correlation to an imminent response to stimulation relative to all other indices

Keywords: antinociception, BIS, general anesthesia, LMA, qCON/qNOX

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5712 The Role of Cholesterol Oxidase of Mycobacterium tuberculosis in the Down-Regulation of TLR2-Signaling Pathway in Human Macrophages during Infection Process

Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

Abstract:

The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

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