Search results for: ethanol tolerance
9 Fuzzy Data, Random Drift, and a Theoretical Model for the Sequential Emergence of Religious Capacity in Genus Homo
Authors: Margaret Boone Rappaport, Christopher J. Corbally
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The ancient ape ancestral population from which living great ape and human species evolved had demographic features affecting their evolution. The population was large, had great genetic variability, and natural selection was effective at honing adaptations. The emerging populations of chimpanzees and humans were affected more by founder effects and genetic drift because they were smaller. Natural selection did not disappear, but it was not as strong. Consequences of the 'population crash' and the human effective population size are introduced briefly. The history of the ancient apes is written in the genomes of living humans and great apes. The expansion of the brain began before the human line emerged. Coalescence times for some genes are very old – up to several million years, long before Homo sapiens. The mismatch between gene trees and species trees highlights the anthropoid speciation processes, and gives the human genome history a fuzzy, probabilistic quality. However, it suggests traits that might form a foundation for capacities emerging later. A theoretical model is presented in which the genomes of early ape populations provide the substructure for the emergence of religious capacity later on the human line. The model does not search for religion, but its foundations. It suggests a course by which an evolutionary line that began with prosimians eventually produced a human species with biologically based religious capacity. The model of the sequential emergence of religious capacity relies on cognitive science, neuroscience, paleoneurology, primate field studies, cognitive archaeology, genomics, and population genetics. And, it emphasizes five trait types: (1) Documented, positive selection of sensory capabilities on the human line may have favored survival, but also eventually enriched human religious experience. (2) The bonobo model suggests a possible down-regulation of aggression and increase in tolerance while feeding, as well as paedomorphism – but, in a human species that remains cognitively sharp (unlike the bonobo). The two species emerged from the same ancient ape population, so it is logical to search for shared traits. (3) An up-regulation of emotional sensitivity and compassion seems to have occurred on the human line. This finds support in modern genetic studies. (4) The authors’ published model of morality's emergence in Homo erectus encompasses a cognitively based, decision-making capacity that was hypothetically overtaken, in part, by religious capacity. Together, they produced a strong, variable, biocultural capability to support human sociability. (5) The full flowering of human religious capacity came with the parietal expansion and smaller face (klinorhynchy) found only in Homo sapiens. Details from paleoneurology suggest the stage was set for human theologies. Larger parietal lobes allowed humans to imagine inner spaces, processes, and beings, and, with the frontal lobe, led to the first theologies composed of structured and integrated theories of the relationships between humans and the supernatural. The model leads to the evolution of a small population of African hominins that was ready to emerge with religious capacity when the species Homo sapiens evolved two hundred thousand years ago. By 50-60,000 years ago, when human ancestors left Africa, they were fully enabled.Keywords: genetic drift, genomics, parietal expansion, religious capacity
Procedia PDF Downloads 3418 The Relationship between Fight-Flight-Freeze System, Level of Expressed Emotion in Family, and Emotion Regulation Difficulties of University Students: Comparison Experienced to Inexperienced Non-Suicidal Self-Injury Students (NSSI)
Authors: Hyojung Shin, Munhee Kweon
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Non-suicide Self Injuri (NSSI) can be defined as the act of an individual who does not intend to die directly and intentionally damaging his or her body tissues. According to a study conducted by the Korean Ministry of Education in 2018, the NSSI is widely spreading among teenagers, with 7.9 percent of all middle school students and 6.4 percent of high school students reporting experience in NSSI. As such, it is understood that the first time of the NSSI is in adolescence. However, the NSSI may not start and stop at a certain time, but may last longer. However, despite the widespread prevalence of NSSI among teenagers, little is known about the process and maintenance of NSSI college students on a continuous development basis. Korea's NSSI research trends are mainly focused on individual internal vulnerabilities (high levels of painful emotions/awareness, lack of pain tolerance) and interpersonal vulnerabilities (poor communication skills and social problem solving), and little studies have been done on individuals' unique characteristics and environmental factors such as substrate or environmental vulnerability factors. In particular, environmental factors are associated with the occurrence of NSSI by acting as a vulnerability factor that can interfere with the emotional control of individuals, whereas individual factors play a more direct role by contributing to the maintenance of NSSI, so it is more important to consider this for personal environmental involvement in NSSI. This study focused on the Fight-Flight-Freeze System as a factor in the defensive avoidance system of Reward Sensitivity in individual factors. Also, Environmental factors include the level of expressed emotion in family. Wedig and Nock (2007) said that if parents with a self-critical cognitive style take the form of criticizing their children, the experience of NSSI increases. The high level of parental criticism is related to the increasing frequency of NSSI acts as well as to serious levels of NSSI. If the normal coping mechanism fails to control emotions, people want to overcome emotional difficulties even through NSSI, and emotional disturbances experienced by individuals within an unsupported social relationship increase vulnerability to NSSI. Based on these theories, this study is to find ways to prevent NSSI and intervene in counseling effectively by verifying the differences between the characteristics experienced NSSI persons and non-experienced NSSI persons. Therefore, the purpose of this research was to examine the relationship of Fight-Flight-Freeze System (FFFS), level of expressed emotion in family and emotion regulation difficulties, comparing those who experienced Non-Suicidal Self-Injury (NSSI) with those who did not experienced Non-Suicidal Self-Injury (NSSI). The data were collected from university students in Seoul Korea and Gyeonggi-do province. 99 subjects were experienced student of NSSI, while 375 were non- experienced student of NSSI. The results of this study are as follows. First, the result of t-test indicated that NSSI attempters showed a significant difference in fight-flight-freeze system, level of expressed emotion and emotion regulation difficulties, compared with non-attempters. Second, fight-flight-freeze system, level of expressed emotion in family and emotion regulation difficulties of NSSI attempters showed a significant difference in correlation. The correlation was significant only freeze system of fight-flight-freeze system, Level of expressed emotion in family and emotion regulation difficulties. Third, freeze system and level of expressed emotion in family predicted emotion regulation difficulties of NSSI attempters. Fight-freeze system and level of expressed emotion in family predicted emotion regulation difficulties of non-NSSI attempters. Lastly, Practical implications for counselors and limitations of this study are discussed.Keywords: fight-flight-freeze system, level of expressed emotion in family, emotion regulation difficulty, non-suicidal self injury
Procedia PDF Downloads 1097 Metal-Organic Frameworks-Based Materials for Volatile Organic Compounds Sensing Applications: Strategies to Improve Sensing Performances
Authors: Claudio Clemente, Valentina Gargiulo, Alessio Occhicone, Giovanni Piero Pepe, Giovanni Ausanio, Michela Alfè
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Volatile organic compound (VOC) emissions represent a serious risk to human health and the integrity of the ecosystems, especially at high concentrations. For this reason, it is very important to continuously monitor environmental quality and develop fast and reliable portable sensors to allow analysis on site. Chemiresistors have become promising candidates for VOC sensing as their ease of fabrication, variety of suitable sensitive materials, and simple sensing data. A chemoresistive gas sensor is a transducer that allows to measure the concentration of an analyte in the gas phase because the changes in resistance are proportional to the amount of the analyte present. The selection of the sensitive material, which interacts with the target analyte, is very important for the sensor performance. The most used VOC detection materials are metal oxides (MOx) for their rapid recovery, high sensitivity to various gas molecules, easy fabrication. Their sensing performance can be improved in terms of operating temperature, selectivity, and detection limit. Metal-organic frameworks (MOFs) have attracted a lot of attention also in the field of gas sensing due to their high porosity, high surface area, tunable morphologies, structural variety. MOFs are generated by the self-assembly of multidentate organic ligands connecting with adjacent multivalent metal nodes via strong coordination interactions, producing stable and highly ordered crystalline porous materials with well-designed structures. However, most MOFs intrinsically exhibit low electrical conductivity. To improve this property, MOFs can be combined with organic and inorganic materials in a hybrid fashion to produce composite materials or can be transformed into more stable structures. MOFs, indeed, can be employed as the precursors of metal oxides with well-designed architectures via the calcination method. The MOF-derived MOx partially preserved the original structure with high surface area and intrinsic open pores, which act as trapping centers for gas molecules, and showed a higher electrical conductivity. Core-shell heterostructures, in which the surface of a metal oxide core is completely coated by a MOF shell, forming a junction at the core-shell heterointerface, can also be synthesized. Also, nanocomposite in which MOF structures are intercalated with graphene related materials can also be produced, and the conductivity increases thanks to the high mobility of electrons of carbon materials. As MOF structures, zinc-based MOFs belonging to the ZIF family were selected in this work. Several Zn-based materials based and/or derived from MOFs were produced, structurally characterized, and arranged in a chemo resistive architecture, also exploring the potentiality of different approaches of sensing layer deposition based on PLD (pulsed laser deposition) and, in case of thermally labile materials, MAPLE (Matrix Assisted Pulsed Laser Evaporation) to enhance the adhesion to the support. The sensors were tested in a controlled humidity chamber, allowing for the possibility of varying the concentration of ethanol, a typical analyte chosen among the VOCs for a first survey. The effect of heating the chemiresistor to improve sensing performances was also explored. Future research will focus on exploring new manufacturing processes for MOF-based gas sensors with the aim to improve sensitivity, selectivity and reduce operating temperatures.Keywords: chemiresistors, gas sensors, graphene related materials, laser deposition, MAPLE, metal-organic frameworks, metal oxides, nanocomposites, sensing performance, transduction mechanism, volatile organic compounds
Procedia PDF Downloads 606 Effects of Delphinidin on Lipid Metabolism in HepG2 Cells and Diet-Induced Obese Mice
Authors: Marcela Parra-Vargas, Ana Sandoval-Rodriguez, Roberto Rodriguez-Echevarria, Jose Dominguez-Rosales, Juan Armendariz-Borunda
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Non-alcoholic fatty liver disease (NAFLD) is characterized by an excess of hepatic lipids, and it is to author’s best knowledge, the most prevalent chronic liver disorder. Anthocyanin-rich food consumption is linked to health benefits in metabolic disorders associated with obesity and NAFLD, although the precise functional role of anthocyanidin delphinidin (Dp) has yet to be established. The aim of this study was to investigate the effect of the Dp in NAFLD metabolic alterations by evaluating prevention or amelioration of hepatic lipid accumulation, as well as molecular mechanisms in two experimental obesity-related models of NALFD. In vitro: HepG2 cells were incubated with sodium palmitate (PA, 1 mM) to induce lipotoxic damage, and concomitantly treated with Dp (180 uM) for 24 h. Subsequently, total lipid accumulation was measured by colorimetric staining with Oil Red O, and total intrahepatic triglycerides were determined by an enzymatic assay. To assess molecular mechanisms, cells were pre-treated with PA for 24 h and then exposed to Dp for 1 h. In vivo: four-week-old male C57BL/6Nhsd mice were allocated in two main groups. Mice were fed with standard diet (control) or high-fat and high-carbohydrate diet (45% fat, HFD) for 16 wk to induce NAFLD. Then HFD was divided into subgroups: one treated orally with Dp (15 mg/kg bw, HFD-Dp) every day for 4 wk, while HFD group treated with vehicle (DMSO). Weight and fasting glucose were recorded weekly, while dietary ingestion was measured daily. Insulin tolerance test was performed at the end of treatment. Liver histology was evaluated with H&E and Masson’s trichrome stain. RT-PCR was used to evaluate gene expression and Western Blot to determine levels of protein in both experimental models. Parametric data were analyzed with one-way ANOVA and Tukey’s post-hoc test. Kruskal-Wallis and Mann-Whitney U test for non-parametric data, and P < 0.5 were considered significant. Dp prevented hepatic lipid accumulation by PA in HepG2 hepatocytes. Furthermore, Dp down-regulated gene expression of SREBP1c, FAS, and CPT1a without modifying AMPK phosphorylation levels. In vivo, Dp oral administration did not ameliorate lipid metabolic alterations raised by HFD. Adiposity, dietary ingestion, fasting glucose, and insulin sensitivity after Dp treatment remained similar to HFD group. Histological analysis showed hepatic damage in HFD groups and no differences between HFD and HFD-Dp groups were found. Hepatic gene expression of ACC and FAS were not altered by HFD. SREBP1c was similar in both HFD and HFD-Dp groups. No significant changes were observed in SREBP1c, ACC, and FAS adipose tissue gene expression by HFD or Dp treatment. Additionally, immunoblotting analysis revealed no changes in pathway SIRT1-LKB-AMPK and PPAR alpha by both HFD groups compared to control. In conclusion, the antioxidant Dp may provoke beneficial effects in the prevention of hepatic lipid accumulation. Nevertheless, the oral dose administrated in mice that simulated the total intake of anthocyanins consumed daily by humans has no effect as a treatment on hepatic lipid metabolic alterations and histological abnormalities associated with exposure to chronic HFD. A healthy lifestyle with regular intake of antioxidants such as anthocyanins may prevent metabolic alterations in NAFLD.Keywords: anthocyanins, antioxidants, delphinidin, non-alcoholic fatty liver disease, obesity
Procedia PDF Downloads 2015 Case Report of a Secretory Carcinoma of the Salivary Gland: Clinical Management Following High-Grade Transformation
Authors: Wissam Saliba, Mandy Nicholson
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Secretory carcinoma (SC) is a rare type of salivary gland cancer. It was first realized as a distinct type of malignancy in 2010and wasinitially termed “mammary analogue secretory carcinoma” because of similarities with secretory breast cancer. The name was later changed to SC. Most SCs originate in parotid glands, and most harbour a rare gene mutation: ETV6-NTRK3. This mutation is rare in common cancers and common in rare cancers; it is present in most secretory carcinomas. Disease outcomes for SC are usually described as favourable as many cases of SC are lowgrade (LG), and cancer growth is slow. In early stages, localized therapy is usually indicated (surgery and/or radiation). Despitea favourable prognosis, a sub-set of casescan be much more aggressive.These cases tend to be of high-grade(HG).HG casesare associated with a poorer prognosis.Management of such cases can be challenging due to limited evidence for effective systemic therapy options. This case report describes the clinical management of a 46-year-oldmale patient with a unique case of SC. He was initially diagnosed with a low/intermediate grade carcinoma of the left parotid gland in 2009; he was treated with surgery and adjuvant radiation. Surgical pathology favoured primary salivary adenocarcinoma, and 2 lymph nodes were positive for malignancy. SC was not yet realized as a distinct type of cancerat the time of diagnosis, and the pathology reportvalidated this gap by stating that the specimen lacked features of the defined types of salivary carcinoma.Slow-growing pulmonary nodules were identified in 2017. In 2020, approximately 11 years after the initial diagnosis, the patient presented with malignant pleural effusion. Pathology from a pleural biopsy was consistent with metastatic poorly differentiated cancer of likely parotid origin, likely mammary analogue secretory carcinoma. The specimen was sent for Next Generation Sequencing (NGS); ETV6-NTRK3 gene fusion was confirmed, and systemic therapy was initiated.One cycle ofcarboplatin/paclitaxel was given in June 2020. He was switched to Larotrectinib (NTRK inhibitor (NTRKi)) later that month. Larotrectinib continued for approximately 9 months, with discontinuation in March 2021 due to disease progression. A second-generation NTRKi (Selitrectinib) was accessed and prescribedthrough a single patient study. Selitrectinib was well tolerated. The patient experienced a complete radiological response within~4 months. Disease progression occurred once again in October 2021. Progression was slow, and Selitrectinib continuedwhile the medical team performed a thorough search for additional treatment options. In January 2022, a liver lesion biopsy was performed, and NGS showed an NTRKG623R solvent-front resistance mutation. Various treatment pathways were considered. The patient pursuedanother investigational NTRKi through a clinical trial, and Selitrectinib was discontinued in July 2022. Excellent performance status was maintained throughout the entire course of treatment.It can be concluded that NTRK inhibitors provided satisfactory treatment efficacy and tolerance for this patient with high-grade transformation and NTRK gene fusion cancer. In the future, more clinical research is needed on systemic treatment options for high-grade transformations in NTRK gene fusion SCs.Keywords: secretory carcinoma, high-grade transformations, NTRK gene fusion, NTRK inhibitor
Procedia PDF Downloads 1074 The Procedural Sedation Checklist Manifesto, Emergency Department, Jersey General Hospital
Authors: Jerome Dalphinis, Vishal Patel
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The Bailiwick of Jersey is an island British crown dependency situated off the coast of France. Jersey General Hospital’s emergency department sees approximately 40,000 patients a year. It’s outside the NHS, with secondary care being free at the point of care. Sedation is a continuum which extends from a normal conscious level to being fully unresponsive. Procedural sedation produces a minimally depressed level of consciousness in which the patient retains the ability to maintain an airway, and they respond appropriately to physical stimulation. The goals of it are to improve patient comfort and tolerance of the procedure and alleviate associated anxiety. Indications can be stratified by acuity, emergency (cardioversion for life-threatening dysrhythmia), and urgency (joint reduction). In the emergency department, this is most often achieved using a combination of opioids and benzodiazepines. Some departments also use ketamine to produce dissociative sedation, a cataleptic state of profound analgesia and amnesia. The response to pharmacological agents is highly individual, and the drugs used occasionally have unpredictable pharmacokinetics and pharmacodynamics, which can always result in progression between levels of sedation irrespective of the intention. Therefore, practitioners must be able to ‘rescue’ patients from deeper sedation. These practitioners need to be senior clinicians with advanced airway skills (AAS) training. It can lead to adverse effects such as dangerous hypoxia and unintended loss of consciousness if incorrectly undertaken; studies by the National Confidential Enquiry into Patient Outcome and Death (NCEPOD) have reported avoidable deaths. The Royal College of Emergency Medicine, UK (RCEM) released an updated ‘Safe Sedation of Adults in the Emergency Department’ guidance in 2017 detailing a series of standards for staff competencies, and the required environment and equipment, which are required for each target sedation depth. The emergency department in Jersey undertook audit research in 2018 to assess their current practice. It showed gaps in clinical competency, the need for uniform care, and improved documentation. This spurred the development of a checklist incorporating the above RCEM standards, including contraindication for procedural sedation and difficult airway assessment. This was approved following discussion with the relevant heads of departments and the patient safety directorates. Following this, a second audit research was carried out in 2019 with 17 completed checklists (11 relocation of joints, 6 cardioversions). Data was obtained from looking at the controlled resuscitation drugs book containing documented use of ketamine, alfentanil, and fentanyl. TrakCare, which is the patient electronic record system, was then referenced to obtain further information. The results showed dramatic improvement compared to 2018, and they have been subdivided into six categories; pre-procedure assessment recording of significant medical history and ASA grade (2 fold increase), informed consent (100% documentation), pre-oxygenation (88%), staff (90% were AAS practitioners) and monitoring (92% use of non-invasive blood pressure, pulse oximetry, capnography, and cardiac rhythm monitoring) during procedure, and discharge instructions including the documented return of normal vitals and consciousness (82%). This procedural sedation checklist is a safe intervention that identifies pertinent information about the patient and provides a standardised checklist for the delivery of gold standard of care.Keywords: advanced airway skills, checklist, procedural sedation, resuscitation
Procedia PDF Downloads 1153 Experimental Proof of Concept for Piezoelectric Flow Harvesting for In-Pipe Metering Systems
Authors: Sherif Keddis, Rafik Mitry, Norbert Schwesinger
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Intelligent networking of devices has rapidly been gaining importance over the past years and with recent advances in the fields of microcontrollers, integrated circuits and wireless communication, low power applications have emerged, enabling this trend even more. Connected devices provide a much larger database thus enabling highly intelligent and accurate systems. Ensuring safe drinking water is one of the fields that require constant monitoring and can benefit from an increased accuracy. Monitoring is mainly achieved either through complex measures, such as collecting samples from the points of use, or through metering systems typically distant to the points of use which deliver less accurate assessments of the quality of water. Constant metering near the points of use is complicated due to their inaccessibility; e.g. buried water pipes, locked spaces, which makes system maintenance extremely difficult and often unviable. The research presented here attempts to overcome this challenge by providing these systems with enough energy through a flow harvester inside the pipe thus eliminating the maintenance requirements in terms of battery replacements or containment of leakage resulting from wiring such systems. The proposed flow harvester exploits the piezoelectric properties of polyvinylidene difluoride (PVDF) films to convert turbulence induced oscillations into electrical energy. It is intended to be used in standard water pipes with diameters between 0.5 and 1 inch. The working principle of the harvester uses a ring shaped bluff body inside the pipe to induce pressure fluctuations. Additionally the bluff body houses electronic components such as storage, circuitry and RF-unit. Placing the piezoelectric films downstream of that bluff body causes their oscillation which generates electrical charge. The PVDF-film is placed as a multilayered wrap fixed to the pipe wall leaving the top part to oscillate freely inside the flow. The warp, which allows for a larger active, consists of two layers of 30µm thick and 12mm wide PVDF layered alternately with two centered 6µm thick and 8mm wide aluminum foil electrodes. The length of the layers depends on the number of windings and is part of the investigation. Sealing the harvester against liquid penetration is achieved by wrapping it in a ring-shaped LDPE-film and welding the open ends. The fabrication of the PVDF-wraps is done by hand. After validating the working principle using a wind tunnel, experiments have been conducted in water, placing the harvester inside a 1 inch pipe at water velocities of 0.74m/s. To find a suitable placement of the wrap inside the pipe, two forms of fixation were compared regarding their power output. Further investigations regarding the number of windings required for efficient transduction were made. Best results were achieved using a wrap with 3 windings of the active layers which delivers a constant power output of 0.53µW at a 2.3MΩ load and an effective voltage of 1.1V. Considering the extremely low power requirements of sensor applications, these initial results are promising. For further investigations and optimization, machine designs are currently being developed to automate the fabrication and decrease tolerance of the prototypes.Keywords: maintenance-free sensors, measurements at point of use, piezoelectric flow harvesting, universal micro generator, wireless metering systems
Procedia PDF Downloads 1922 Computational, Human, and Material Modalities: An Augmented Reality Workflow for Building form Found Textile Structures
Authors: James Forren
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This research paper details a recent demonstrator project in which digital form found textile structures were built by human craftspersons wearing augmented reality (AR) head-worn displays (HWDs). The project utilized a wet-state natural fiber / cementitious matrix composite to generate minimal bending shapes in tension which, when cured and rotated, performed as minimal-bending compression members. The significance of the project is that it synthesizes computational structural simulations with visually guided handcraft production. Computational and physical form-finding methods with textiles are well characterized in the development of architectural form. One difficulty, however, is physically building computer simulations: often requiring complicated digital fabrication workflows. However, AR HWDs have been used to build a complex digital form from bricks, wood, plastic, and steel without digital fabrication devices. These projects utilize, instead, the tacit knowledge motor schema of the human craftsperson. Computational simulations offer unprecedented speed and performance in solving complex structural problems. Human craftspersons possess highly efficient complex spatial reasoning motor schemas. And textiles offer efficient form-generating possibilities for individual structural members and overall structural forms. This project proposes that the synthesis of these three modalities of structural problem-solving – computational, human, and material - may not only develop efficient structural form but offer further creative potentialities when the respective intelligence of each modality is productively leveraged. The project methodology pertains to its three modalities of production: 1) computational, 2) human, and 3) material. A proprietary three-dimensional graphic statics simulator generated a three-legged arch as a wireframe model. This wireframe was discretized into nine modules, three modules per leg. Each module was modeled as a woven matrix of one-inch diameter chords. And each woven matrix was transmitted to a holographic engine running on HWDs. Craftspersons wearing the HWDs then wove wet cementitious chords within a simple falsework frame to match the minimal bending form displayed in front of them. Once the woven components cured, they were demounted from the frame. The components were then assembled into a full structure using the holographically displayed computational model as a guide. The assembled structure was approximately eighteen feet in diameter and ten feet in height and matched the holographic model to under an inch of tolerance. The construction validated the computational simulation of the minimal bending form as it was dimensionally stable for a ten-day period, after which it was disassembled. The demonstrator illustrated the facility with which computationally derived, a structurally stable form could be achieved by the holographically guided, complex three-dimensional motor schema of the human craftsperson. However, the workflow traveled unidirectionally from computer to human to material: failing to fully leverage the intelligence of each modality. Subsequent research – a workshop testing human interaction with a physics engine simulation of string networks; and research on the use of HWDs to capture hand gestures in weaving seeks to develop further interactivity with rope and chord towards a bi-directional workflow within full-scale building environments.Keywords: augmented reality, cementitious composites, computational form finding, textile structures
Procedia PDF Downloads 1751 Ultra-Rapid and Efficient Immunomagnetic Separation of Listeria Monocytogenes from Complex Samples in High-Gradient Magnetic Field Using Disposable Magnetic Microfluidic Device
Authors: L. Malic, X. Zhang, D. Brassard, L. Clime, J. Daoud, C. Luebbert, V. Barrere, A. Boutin, S. Bidawid, N. Corneau, J. Farber, T. Veres
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The incidence of infections caused by foodborne pathogens such as Listeria monocytogenes (L. monocytogenes) poses a great potential threat to public health and safety. These issues are further exacerbated by legal repercussions due to “zero tolerance” food safety standards adopted in developed countries. Unfortunately, a large number of related disease outbreaks are caused by pathogens present in extremely low counts currently undetectable by available techniques. The development of highly sensitive and rapid detection of foodborne pathogens is therefore crucial, and requires robust and efficient pre-analytical sample preparation. Immunomagnetic separation is a popular approach to sample preparation. Microfluidic chips combined with external magnets have emerged as viable high throughput methods. However, external magnets alone are not suitable for the capture of nanoparticles, as very strong magnetic fields are required. Devices that incorporate externally applied magnetic field and microstructures of a soft magnetic material have thus been used for local field amplification. Unfortunately, very complex and costly fabrication processes used for integration of soft magnetic materials in the reported proof-of-concept devices would prohibit their use as disposable tools for food and water safety or diagnostic applications. We present a sample preparation magnetic microfluidic device implemented in low-cost thermoplastic polymers using fabrication techniques suitable for mass-production. The developed magnetic capture chip (M-chip) was employed for rapid capture and release of L. monocytogenes conjugated to immunomagnetic nanoparticles (IMNs) in buffer and beef filtrate. The M-chip relies on a dense array of Nickel-coated high-aspect ratio pillars for capture with controlled magnetic field distribution and a microfluidic channel network for sample delivery, waste, wash and recovery. The developed Nickel-coating process and passivation allows generation of switchable local perturbations within the uniform magnetic field generated with a pair of permanent magnets placed at the opposite edges of the chip. This leads to strong and reversible trapping force, wherein high local magnetic field gradients allow efficient capture of IMNs conjugated to L. monocytogenes flowing through the microfluidic chamber. The experimental optimization of the M-chip was performed using commercially available magnetic microparticles and fabricated silica-coated iron-oxide nanoparticles. The fabricated nanoparticles were optimized to achieve the desired magnetic moment and surface functionalization was tailored to allow efficient capture antibody immobilization. The integration, validation and further optimization of the capture and release protocol is demonstrated using both, dead and live L. monocytogenes through fluorescence microscopy and plate- culture method. The capture efficiency of the chip was found to vary as function of listeria to nanoparticle concentration ratio. The maximum capture efficiency of 30% was obtained and the 24-hour plate-culture method allowed the detection of initial sample concentration of only 16 cfu/ml. The device was also very efficient in concentrating the sample from a 10 ml initial volume. Specifically, 280% concentration efficiency was achieved in 17 minutes only, demonstrating the suitability of the system for food safety applications. In addition, flexible design and low-cost fabrication process will allow rapid sample preparation for applications beyond food and water safety, including point-of-care diagnosis.Keywords: array of pillars, bacteria isolation, immunomagnetic sample preparation, polymer microfluidic device
Procedia PDF Downloads 279