Search results for: social data analysis

Authors: Jason Ceavers, Travis Thompson, Juan Torres, Ramanakumar Anam, Alan Wong, Andrei Carvalho, Shane Quo, Shawn Alonso, Moises Cintron, Ricardo C. Carrero, German Lopez, Vamsi Garimella, German Giese

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Introduction: Advance directives (AD) are essential for patients to communicate their wishes when they are not able to. Ideally, these discussions should not occur for the first time when a patient is hospitalized with an acute life-threatening illness. There is a large number of patients who do not have clearly documented ADs, resulting in the misutilization of resources and additional patient harm. This is a nationwide issue, and the Joint Commission has it as one of its national quality metrics. Presented here is a proposed protocol to increase the number of documented AD discussions in a community-based, internal medicine residency primary care clinic in South Florida. Methods: The SMART Aim for this quality improvement project is to increase documentation of AD discussions in the outpatient setting by 25% within three months in medicare patients. A survey was sent to stakeholders (clinic attendings, residents, medical assistants, front desk staff, and clinic managers), asking them for three factors they believed contributed most to the low documentation rate of AD discussions. The two most important factors were time constraints and systems issues (such as lack of a standard method to document ADs and ADs not being uploaded to the chart) which were brought up by 25% and 21.2% of the 32 survey responders, respectively. Pre-intervention data from clinic patients in 2020-2021 revealed 17.05% of patients had clear, actionable ADs documented. To address these issues, an AD pocket card was created to give to patients. One side of the card has a brief explanation of what ADs are. The other side has a column of interventions (cardiopulmonary resuscitation, mechanical ventilation, dialysis, tracheostomy, feeding tube) with boxes patients check off if they want the intervention done, do not want the intervention, do not want to discuss the topic, or need more information. These cards are to be filled out and scanned into their electronic chart to be reviewed by the resident before their appointment. The interventions that patients want more information on will be discussed by the provider. If any changes are made, the card will be re-scanned into their chart. After three months, we will chart review the patients seen in the clinic to determine how many medicare patients have a pocket card uploaded and how many have advance directives discussions documented in a progress note or annual wellness note. If there is not enough time for an AD discussion, a follow-up appointment can be scheduled for that discussion. Discussion: ADs are a crucial part of patient care, and failure to understand a patient’s wishes leads to improper utilization of resources, avoidable litigation, and patient harm. Time constraints and systems issues were identified as two major factors contributing to the lack of advance directive discussion in our community-based resident primary care clinic. Our project aims at increasing the documentation rate for ADs through a simple pocket card intervention. These are self-explanatory, easy to read and allow the patients to clearly express what interventions they desire or what they want to discuss further with their physician.

Keywords: advance directives, community-based, pocket card, primary care clinic

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Authors: Kaustav Mukherjee

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In this paper we propose a Neural Style Transfer solution whereby we have created a Lightweight Separable Convolution Kernel Based GAN Architecture (SC-GAN) which will very useful for designing filter for Mobile Phone Cameras and also Edge Devices which will convert any image to its 2D ANIMATED COMIC STYLE Movies like HEMAN, SUPERMAN, JUNGLE-BOOK. This will help the 2D animation artist by relieving to create new characters from real life person's images without having to go for endless hours of manual labour drawing each and every pose of a cartoon. It can even be used to create scenes from real life images.This will reduce a huge amount of turn around time to make 2D animated movies and decrease cost in terms of manpower and time. In addition to that being extreme light-weight it can be used as camera filters capable of taking Comic Style Shots using mobile phone camera or edge device cameras like Raspberry Pi 4,NVIDIA Jetson NANO etc. Existing Methods like CartoonGAN with the model size close to 170 MB is too heavy weight for mobile phones and edge devices due to their scarcity in resources. Compared to the current state of the art our proposed method which has a total model size of 31 MB which clearly makes it ideal and ultra-efficient for designing of camera filters on low resource devices like mobile phones, tablets and edge devices running OS or RTOS. .Owing to use of high resolution input and usage of bigger convolution kernel size it produces richer resolution Comic-Style Pictures implementation with 6 times lesser number of parameters and with just 25 extra epoch trained on a dataset of less than 1000 which breaks the myth that all GAN need mammoth amount of data. Our network reduces the density of the Gan architecture by using Depthwise Separable Convolution which does the convolution operation on each of the RGB channels separately then we use a Point-Wise Convolution to bring back the network into required channel number using 1 by 1 kernel.This reduces the number of parameters substantially and makes it extreme light-weight and suitable for mobile phones and edge devices. The architecture mentioned in the present paper make use of Parameterised Batch Normalization Goodfellow etc al. (Deep Learning OPTIMIZATION FOR TRAINING DEEP MODELS page 320) which makes the network to use the advantage of Batch Norm for easier training while maintaining the non-linear feature capture by inducing the learnable parameters

Keywords: comic stylisation from camera image using GAN, creating 2D animated movie style custom stickers from images, depth-wise separable convolutional neural network for light-weight GAN architecture for EDGE devices, GAN architecture for 2D animated cartoonizing neural style, neural style transfer for edge, model distilation, perceptual loss

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Authors: Haresh Sharma, Jaimini Luharia

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The ecology in the Western Himalayan region transforms with the change in altitude. This change is observed in terms of topography, species of flora and fauna and the quality of the soil. The current study focuses on women of indigenous communities of Pangi Valley, which is located in the state of Himachal Pradesh, India. The valley is bifurcated into three different areas –Saichu, Hudan Bhatori, and Sural Bhatori valleys. It is one of the most remote, rugged and difficult to access tribal regions of Chamba district. The altitude of the valley ranges from 2,000 m to 6,000 m above sea level. The Pangi valley is inhabited by ‘Pangwals’ and ‘Bhots’ tribes of the Himalayas who speak their local tribal language called’ Pangwali’. The valley is cut-off from the mainland due to heavy snow and lack of proper roads during peak winters. Due to difficult geographical location, the daily lives of the people are constantly challenged, and they are most of the times deprived of benefits targeted through government programs. However, the indigenous communities earn their livelihood through livestock and forest-based produce while some of them migrate to nearby places for better work. The current study involves snowball sampling methodology for data collection along with in-depth interviews of women members of Self-Help Groups and women farmers. The findings reveal that the lives of these indigenous communities largely depend on forest-based products. So, it creates all the more significance of enhancing, maintaining, and consuming natural resources sustainably. Under such circumstances, the women of the community play a significant role of guardians in conservation and protection of the forests. They are the custodians of traditional knowledge of environment conservation practices that have been followed for many years in the region. The present study also sought to establish a relationship between some of the development initiatives undertaken by the women in the valley that stimulate sustainable mountain economy and conservation practices. These initiatives include cultivation of products like hazelnut, ‘Gucchi’ rare quality mushroom, medicinal plants exclusively found in the region, thereby promoting long term sustainable conservation of agro-biodiversity of the Western Himalayan region. The measures taken by the community women are commendable as they ensure access and distribution of natural resources as well as manage them for future generations. Apart from this, the tribal women have actively formed Self-Help Groups promoting financial inclusion through various activities that augment ownership and accountability towards the overall development of the communities. But, the results also suggest that there’s not enough recognition given to women’s role in forests conservation practices due to several local socio-political reasons. There are not enough research studies done on communities of Pangi Valley due to inaccessibility created out of lack of proper roads and other resources. Also, there emerged a need to concretize indigenous and traditional knowledge of conservation practices followed by women in the community.

Keywords: forest conservation, indigenous community women, sustainable livelihoods, sustainable development, poverty alleviation, Western Himalayas

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Authors: Eman Hegazy, Stefan Anderberg, Joakim Krook

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Urban water infrastructure is crucial for efficient and reliable water supply in growing cities. With the growth of cities, the need for maintenance and renewal of these systems increases but often goes unfulfilled due to a variety of reasons, such as limited funding, political priorities, or lack of public awareness. Neglecting the renewal needs of these systems can lead to frequent malfunctions and reduced quality and reliability of water supply, as well as increased costs and health and environmental hazards. It is important for cities to prioritize investment in water infrastructure and develop long-term plans to address renewal needs. Drawing general conclusions about the rate of renewal of urban water infrastructure systems at an international or national level can be challenging due to the influence of local management decisions. In many countries, the responsibility for water infrastructure management lies with the municipal authorities, who are responsible for making decisions about the allocation of resources for repair, maintenance, and renewal. These decisions can vary widely based on factors such as local finances, political priorities, and public perception of the importance of water infrastructure. As a result, it is difficult to make generalizations about the rate of renewal across different countries or regions. In Sweden, the situation is not different, and the information from Svenskt Vatten indicates that the rate of renewal varies across municipalities and can be insufficient, leading to a buildup of maintenance and renewal needs. This study aims to examine the adequacy of the rate of renewal of urban water infrastructure in Linköping case city in Sweden. Using a case study framework, the study will assess the current status of the urban water system and the need for renewal. The study will also consider the role of factors such as proper identification processes, limited funding, competing for political priorities, and local management decisions in contributing to insufficient renewal. The study investigates the following questions: (1) What is the current status of water and sewerage networks in terms of length, age distribution, and material composition, estimated total water leakage in the network per year, damages, leaks, and outages occur per year, both overall and by district? (2) What are the main causes of these damages, leaks, and interruptions, and how are they related to lack of maintenance and renewal? (3) What is the current status of renewal work for the water and sewerage networks, including the renewal rate and changes over time, recent renewal material composition, and the budget allocation for renewal and emergency repairs? (4) What factors influence the need for renewal and what conditions should be considered in the assessment? The findings of the study provide insights into the challenges facing urban water infrastructure and identify strategies for improving the rate of renewal to ensure a reliable and sustainable water supply.

Keywords: case study, infrastructure, management, renewal need, Sweden

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Authors: Aurica P. Chiriac, Vera Balan, Mihai Asandulesa, Elena Butnaru, Nita Tudorachi, Elena Stoleru, Loredana E. Nita, Iordana Neamtu, Alina Diaconu, Liliana Mititelu-Tartau

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The interest for studying ‘smart’ materials is entirely justified and in this context were realized investigations on poly(2-hydroxyethylmethacrylate-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane), which is a macromolecular compound with sensibility at pH and temperature, gel formation capacity, binding properties, amphilicity, good oxidative and thermal stability. Physico-chemical characteristics in terms of the molecular weight, temperature-sensitive abilities and thermal stability, as well rheological, dielectric and spectroscopic properties were evaluated in correlation with further coupling capabilities. Differential scanning calorimetry investigation indicated Tg at 36.6 °C and a melting point at Tm=72.8°C, for the studied copolymer, and up to 200oC two exothermic processes (at 99.7°C and 148.8°C) were registered with losing weight of about 4 %, respective 19.27%, which indicate just processes of thermal decomposition (and not phenomena of thermal transition) owing to scission of the functional groups and breakage of the macromolecular chains. At the same time, the rheological studies (rotational tests) confirmed the non-Newtonian shear-thinning fluid behavior of the copolymer solution. The dielectric properties of the copolymer have been evaluated in order to investigate the relaxation processes and two relaxation processes under Tg value were registered and attributed to localized motions of polar groups from side chain macromolecules, or parts of them, without disturbing the main chains. According to literature and confirmed as well by our investigations, β-relaxation is assigned with the rotation of the ester side group and the γ-relaxation corresponds to the rotation of hydroxy- methyl side groups. The fluorescence spectroscopy confirmed the copolymer structure, the spiroacetal moiety getting an axial conformation, more stable, with lower energy, able for specific interactions with molecules from environment, phenomena underlined by different shapes of the emission spectra of the copolymer. Also, the copolymer was used as template for indomethacin incorporation as model drug, and the biocompatible character of the complex was confirmed. The release behavior of the bioactive compound was dependent by the copolymer matrix composition, the increasing of 3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane comonomer amount attenuating the drug release. At the same time, the in vivo studies did not show significant differences of leucocyte formula elements, GOT, GPT and LDH levels, nor immune parameters (OC, PC, and BC) between control mice group and groups treated just with copolymer samples, with or without drug, data attesting the biocompatibility of the polymer samples. The investigation of the physico-chemical characteristics of poly(2-hydrxyethyl methacrylate-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane) in terms of temperature-sensitive abilities, rheological and dielectrical properties, are bringing useful information for further specific use of this polymeric compound.

Keywords: bioapplications, dielectric and spectroscopic properties, dual sensitivity at pH and temperature, smart materials

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Authors: Tracie Catlett

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Students enrolled in high-achieving schools are under tremendous pressure to perform at high levels inside and outside the classroom. Achievement pressure is a prevalent source of stress for students enrolled in high-achieving schools, and female students in particular experience a higher frequency and higher levels of stress compared to their male peers. The practice of mindfulness in a school setting is one tool that has been linked to improved self-regulation of emotions, increased positive emotions, and stress reduction. A mixed methods randomized pretest-posttest no-treatment control trial, evaluated the effects of a six-session mindfulness intervention taught during a regularly scheduled life skills period in an independent day school, one type of high-achieving school. Twenty-nine students in Grades 10 and 11 were randomized by class where Grade 11 students were in the intervention group (n = 14) and Grade 10 students were in the control group (n = 15). Findings from the study produced mixed results. There was no evidence that the mindfulness program reduced participants’ stress levels and negative emotions. In fact, contrary to what was expected, students enrolled in the intervention group experienced higher levels of stress and increased negative emotions at posttreatment when compared to pretreatment. Neither the within-group nor the between-groups changes in stress level were statistically significant, p > .05, and the between-groups effect size was small, d = .2. The study found evidence that the mindfulness program may have had a positive impact on students’ ability to regulate their emotions. The within-group comparison and the between-groups comparison at posttreatment found that students in the mindfulness course experienced statistically significant improvement in the in their ability to regulate their emotions at posttreatment, p = .009 < .05 and p =. 034 < .05, respectively. The between-groups effect size was medium, d =.7, suggesting that the positive differences in emotion regulation difficulties were substantial and have practical implications. The analysis of gender differences as they relate to stress and emotions revealed that female students perceive higher levels of stress and report experiencing stress more often than males. There were no gender differences when analyzing sources of stress experienced by the student participants. Both females and males experience regular achievement pressures related to their school performance and worry about their future, college acceptance, grades, and parental expectations. Females reported an increased awareness of their stress and actively engaged in practicing mindfulness to manage their stress. Students in the treatment group expressed that the practice of mindfulness resulted in feelings of relaxation and calmness.

Keywords: achievement pressure, adolescents, emotion regulation, emotions, high-achieving schools, independent schools, mindfulness, negative affect, positive affect, stress

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Authors: Peter Kus, Anna Ostroverkh, Yurii Yakovlev, Yevheniia Lobko, Roman Fiala, Ivan Khalakhan, Vladimir Matolin

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Hydrogen economy is a concept of low-emission society which harvests most of its energy from renewable sources (e.g., wind and solar) and in case of overproduction, electrochemically turns the excess amount into hydrogen, which serves as an energy carrier. Proton exchange membrane water electrolyzers (PEMWE) are the backbone of this concept. By fast-response electricity to hydrogen conversion, the PEMWEs will not only stabilize the electrical grid but also provide high-purity hydrogen for variety of fuel cell powered devices, ranging from consumer electronics to vehicles. Wider commercialization of PEMWE technology is however hindered by high prices of noble metals which are necessary for catalyzing the redox reactions within the cell. Namely, platinum for hydrogen evolution reaction (HER), running on cathode, and iridium for oxygen evolution reaction (OER) on anode. Possible way of how to lower the loading of Pt and Ir is by using conductive high-surface nanostructures as catalyst supports in conjunction with thin-film catalyst deposition. The presented study discusses unconventional technique of membrane electron assembly (MEA) preparation. Noble metal catalysts (Pt and Ir) were magnetron sputtered in very low loadings onto the surface of porous sublayers (located on gas diffusion layer or directly on membrane), forming so to say localized three-phase boundary. Ultrasonically sprayed corrosion resistant TiC-based sublayer was used as a support material on anode, whereas magnetron sputtered nanostructured etched nitrogenated carbon (CNx) served the same role on cathode. By using this configuration, we were able to significantly decrease the amount of noble metals (to thickness of just tens of nanometers), while keeping the performance comparable to that of average state-of-the-art catalysts. Complex characterization of prepared supported catalysts includes in-cell performance and durability tests, electrochemical impedance spectroscopy (EIS) as well as scanning electron microscopy (SEM) imaging and X-ray photoelectron spectroscopy (XPS) analysis. Our research proves that magnetron sputtering is a suitable method for thin-film deposition of electrocatalysts. Tested set-up of thin-film supported anode and cathode catalysts with combined loading of just 120 ug.cm⁻² yields remarkable values of specific current. Described approach of thin-film low-loading catalyst deposition might be relevant when noble metal reduction is the topmost priority.

Keywords: hydrogen economy, low-loading catalyst, magnetron sputtering, proton exchange membrane water electrolyzer

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Authors: M. Vezir Kahraman, Emre Basturk

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Energy storage technology offers new ways to meet the demand to obtain efficient and reliable energy storage materials. Thermal energy storage systems provide the potential to acquire energy savings, which in return decrease the environmental impact related to energy usage. For this purpose, phase change materials (PCMs) that work as 'latent heat storage units' which can store or release large amounts of energy are preferred. Phase change materials (PCMs) are being utilized to absorb, collect and discharge thermal energy during the cycle of melting and freezing, converting from one phase to another. Phase Change Materials (PCMs) can generally be arranged into three classes: organic materials, salt hydrates and eutectics. Many kinds of organic and inorganic PCMs and their blends have been examined as latent heat storage materials. PCMs have found different application areas such as solar energy storage and transfer, HVAC (Heating, Ventilating and Air Conditioning) systems, thermal comfort in vehicles, passive cooling, temperature controlled distributions, industrial waste heat recovery, under floor heating systems and modified fabrics in textiles. Ultraviolet (UV)-curing technology has many advantages, which made it applicable in many different fields. Low energy consumption, high speed, room-temperature operation, low processing costs, high chemical stability, and being environmental friendly are some of its main benefits. UV-curing technique has many applications. One of the many advantages of UV-cured PCMs is that they prevent the interior PCMs from leaking. Shape-stabilized PCM is prepared by blending the PCM with a supporting material, usually polymers. In our study, this problem is minimized by coating the fatty alcohols with a photo-cross-linked thiol-ene based polymeric system. Leakage is minimized because photo-cross-linked polymer acts a matrix. The aim of this study is to introduce a novel thiol-ene based shape-stabilized PCM. Photo-crosslinked thiol-ene based polymers containing fatty alcohols were prepared and characterized for the purpose of phase change materials (PCMs). Different types of fatty alcohols were used in order to investigate their properties as shape-stable PCMs. The structure of the PCMs was confirmed by ATR-FTIR techniques. The phase transition behaviors, thermal stability of the prepared photo-crosslinked PCMs were investigated by differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). This work was supported by Marmara University, Commission of Scientific Research Project.

Keywords: differential scanning calorimetry (DSC), Polymeric phase change material, thermal energy storage, UV-curing

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Authors: Heitor Oliveira, Gabriele De-Waal, Juergen Schmenger, Lynsey Godfrey, Tibor Kovacs

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Rheolaser MASTER™ makes use of multiple scattering of light, caused by scattering objects in a continuous medium (such as droplets and particles in colloids), to characterize the viscoelasticity of soft materials. It offers an alternative to conventional rheometers to characterize viscoelasticity of products such as hair cosmetics. Up to six simultaneous measurements at controlled temperature can be carried out simultaneously (10-15 min), and the method requires only minor sample preparation work. Conversely to conventional rheometer based methods, no mechanical stress is applied to the material during the measurements. Therefore, the properties of the exact same sample can be monitored over time, like in aging and stability studies. We determined the elastic index (EI) of water/emulsion mixtures (1 ≤ fat alcohols (FA) ≤ 5 wt%) and emulsion/gel-network mixtures (8 ≤ FA ≤ 17 wt%) and compared with the elastic/sorage mudulus (G’) for the respective samples using a TA conventional rheometer with flat plates geometry. As expected, it was found that log(EI) vs log(G’) presents a linear behavior. Moreover, log(EI) increased in a linear fashion with solids level in the entire range of compositions (1 ≤ FA ≤ 17 wt%), while rheometer measurements were limited to samples down to 4 wt% solids level. Alternatively, a concentric cilinder geometry would be required for more diluted samples (FA > 4 wt%) and rheometer results from different sample holder geometries are not comparable. The plot of the rheolaser output parameters solid-liquid balance (SLB) vs EI were suitable to monitor product aging processes. These data could quantitatively describe some observations such as formation of lumps over aging time. Moreover, this method allowed to identify that the different specifications of a key raw material (RM < 0.4 wt%) in the respective gel-network (GN) product has minor impact on product viscoelastic properties and it is not consumer perceivable after a short aging time. Broadening of a RM spec range typically has a positive impact on cost savings. Last but not least, the photon path length (λ*)—proportional to droplet size and inversely proportional to volume fraction of scattering objects, accordingly to the Mie theory—and the EI were suitable to characterize product destabilization processes (e.g., coalescence and creaming) and to predict product stability about eight times faster than our standard methods. Using these parameters we could successfully identify formulation and process parameters that resulted in unstable products. In conclusion, Rheolaser allows quick and reliable characterization of viscoelastic properties of hair cosmetics that are related to their performance and stability. It operates in a broad range of product compositions and has applications spanning from the formulation of our hair cosmetics to fast release criteria in our production sites. Last but not least, this powerful tool has positive impact on R&D development time—faster delivery of new products to the market—and consequently on cost savings.

Keywords: colloids, hair cosmetics, light scattering, performance and stability, soft materials, viscoelastic properties

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Authors: Jin Sheng

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This paper addresses a promising area of insurance sector to develop in Asia. Captive insurance, which provides risk-mitigation services for its parent company, has great potentials to develop in energy, infrastructure, agriculture, logistics, catastrophe, and alternative risk transfer (ART), and will greatly affect the framework of insurance industry. However, the Asian captive insurance market only takes a small proportion in the global market. The recent supply chain interruption case of Hanjin Shipping indicates the significance of risk management for an Asian company’s sustainability and resilience. China has substantial needs and great potentials to develop captive insurance, on account of the currency volatility, enterprises’ credit risks, and legal and operational risks of the Belt and Road initiative. Up to date, Mainland Chinese enterprises only have four offshore captives incorporated by CNOOC, Sinopec, Lenovo and CGN Power), three onshore captive insurance companies incorporated by CNPC, China Railway, and COSCO, as well as one industrial captive insurance organization - China Ship-owners Mutual Assurance Association. Its captive market grows slowly with one or two captive insurers licensed yearly after September 2011. As an international financial center, Hong Kong has comparative advantages in taxation, professionals, market access and well-established financial infrastructure to develop a functional captive insurance market. For example, Hong Kong’s income tax for an insurance company is 16.5%; while China's income tax for an insurance company is 25% plus business tax of 5%. Furthermore, restrictions on market entry and operations of China’s onshore captives make establishing offshore captives in international or regional captive insurance centers such as Singapore, Hong Kong, and other overseas jurisdictions to become attractive options. Thus, there are abundant business opportunities in this area. Using methodology of comparative studies and case analysis, this paper discusses the incorporation, regulatory issues, taxation and prospect of captive insurance market in Hong Kong, China and Singapore. Hong Kong and Singapore are both international financial centers with prominent advantages in tax concessions, technology, implementation, professional services, and well-functioning legal system. Singapore, as the domicile of 71 active captives, has been the largest captive insurance hub in Asia, as well as an established reinsurance hub. Hong Kong is an emerging captive insurance hub with 5 to 10 newly licensed captives each year, according to the Hong Kong Financial Services Development Council. It is predicted that Hong Kong will become a domicile for 50 captive insurers by 2025. This paper also compares the formation of a captive in Singapore with other jurisdictions such as Bermuda and Vermont.

Keywords: Alternative Risk Transfer (ART), captive insurance company, offshore captives, risk management, reinsurance, self-insurance fund

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Authors: Dagmara A. Niedziela, Mark P. Murphy, Orla M. Keane, Finola C. Leonard

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Staphylococcus aureus is the most frequent cause of clinical and subclinical bovine mastitis in Ireland. Mastitis caused by S. aureus is often chronic and tends to recur after antibiotic treatment. This may be due to several virulence factors, including attributes that enable the bacterium to internalize into bovine mammary epithelial cells, where it may evade antibiotic treatment, or evade the host immune response. Four bovine-adapted lineages (CC71, CC97, CC151 and ST136) were identified among a collection of Irish S. aureus mastitis isolates. Genotypic variation of mastitis-causing strains may contribute to different presentations of the disease, including differences in milk somatic cell count (SCC), the main method of mastitis detection. The objective of this study was to investigate the influence of bacterial strain and lineage on host immune response, by employing cell culture methods in vitro as well as an in vivo infection model. Twelve bovine adapted S. aureus strains were examined for internalization into bovine mammary epithelial cells (bMEC) and their ability to induce an immune response from bMEC (using qPCR and ELISA). In vitro studies found differences in a variety of virulence traits between the lineages. Strains from lineages CC97 and CC71 internalized more efficiently into bovine mammary epithelial cells (bMEC) than CC151 and ST136. CC97 strains also induced immune genes in bMEC more strongly than strains from the other 3 lineages. One strain each of CC151 and CC97 that differed in their ability to cause an immune response in bMEC were selected on the basis of the above in vitro experiments. Fourteen first-lactation Holstein-Friesian cows were purchased from 2 farms on the basis of low SCC (less than 50 000 cells/ml) and infection free status. Seven cows were infected with 1.73 x 102 c.f.u. of the CC97 strain (Group 1) and another seven with 5.83 x 102 c.f.u. of the CC151 strain (Group 2). The contralateral quarter of each cow was inoculated with PBS (vehicle). Clinical signs of infection (temperature, milk and udder appearance, milk yield) were monitored for 30 days. Blood and milk samples were taken to determine bacterial counts in milk, SCC, white blood cell populations and cytokines. Differences in disease presentation in vivo between groups were observed, with two animals from Group 2 developing clinical mastitis and requiring antibiotic treatment, while one animal from Group 1 did not develop an infection for the duration of the study. Fever (temperature > 39.5⁰C) was observed in 3 animals from Group 2 and in none from Group 1. Significant differences in SCC and bacterial load between groups were observed in the initial stages of infection (week 1). Data is also being collected on cytokines and chemokines secreted during the course of infection. The results of this study suggest that a strain from lineage CC151 may cause more severe clinical mastitis, while a strain from lineage CC97 may cause mild, subclinical mastitis. Diversity between strains of S. aureus may therefore influence the clinical presentation of mastitis, which in turn may influence disease detection and treatment needs.

Keywords: Bovine mastitis, host immune response, host-pathogen interactions, Staphylococcus aureus

Procedia PDF Downloads 157

Authors: Thaís T. Valério Caetano, Lívia de Carvalho Ferreira, João Máximo De Siqueira, Carlos Alexandre Carollo, Arthur Ladeira Macedo, Vanessa C. Stein

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Nicotiana glauca, commonly known as wild tobacco or tobacco bush, belongs to the Solanaceae family. It is native to South America but has become naturalized in various regions, including Australia, California, Africa, and the Mediterranean. N. glauca is listed in the Global Invasive Species Database (GISD) and the Invasive Species Compendium (CABI). It is known for producing pyridine alkaloids, including anabasine, which is highly toxic. Anabasine is predominantly found in the leaves and can cause severe health issues such as neuromuscular blockade, respiratory arrest, and cardiovascular problems when ingested. Mistaken identity with edible plants like spinach has resulted in food poisoning cases in Israel and Brazil. Anabasine, a minor alkaloid constituent of tobacco, may contribute to tobacco addiction by mimicking or enhancing the effects of nicotine. Therefore, it is essential to investigate the production pattern of anabasine and its relationship to the developmental stages of the plant. This study aimed to establish the relationship between the phenological plant age, cultivation place, and the increase in anabasine concentration, which can lead to human intoxication cases. In this study, N. glauca plants were collected from three different rural areas in Brazil during a year to examine leaves at various stages of development. Samples were also obtained from cultivated plants in Marilândia, Minas Gerais, Brazil, as well as from Divinópolis, Minas Gerais, Brazil, and Arraial do Cabo, Rio de Janeiro, Brazil. In vitro cultivated plants on MS medium were included in the study. The collected leaves were dried, powdered, and stored. Alkaloid extraction was performed using a methanol and water mixture, followed by liquid-liquid extraction with chloroform. The anabasine content was determined using HPLC-DAD analysis with nicotine as a standard. The results indicated that anabasine production increases with the plant's development, peaking in adult leaves during the reproduction phase and declining afterward. In vitro, plants showed similar anabasine production to young leaves. The successful adaptation of N. glauca in new environments poses a global problem, and the correlation between anabasine production and the plant's developmental stages has been understudied. The presence of substances produced by the plant can pose a risk to other species, especially when mistaken for edible plants. The findings from this study shed light on the pattern of anabasine production and its association with plant development, contributing to a better understanding of the potential risks associated with N. glauca and the importance of accurate identification.

Keywords: alkaloid production, invasive species, nicotiana glauca, plant phenology

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Authors: J. J. Rickard, A. Belli, P. Goldberg Oppenheimer

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Medical diagnostics, environmental monitoring, homeland security and forensics increasingly demand specific and field-deployable analytical technologies for quick point-of-care diagnostics. Although technological advancements have made optical methods well-suited for miniaturization, a highly-sensitive detection technique for minute sample volumes is required. Raman spectroscopy is a well-known analytical tool, but has very weak signals and hence is unsuitable for trace level analysis. Enhancement via localized optical fields (surface plasmons resonances) on nanoscale metallic materials generates huge signals in surface-enhanced Raman scattering (SERS), enabling single molecule detection. This enhancement can be tuned by manipulation of the surface roughness and architecture at the sub-micron level. Nevertheless, the development and application of SERS has been inhibited by the irreproducibility and complexity of fabrication routes. The ability to generate straightforward, cost-effective, multiplex-able and addressable SERS substrates with high enhancements is of profound interest for SERS-based sensing devices. While most SERS substrates are manufactured by conventional lithographic methods, the development of a cost-effective approach to create nanostructured surfaces is a much sought-after goal in the SERS community. Here, a method is established to create controlled, self-organized, hierarchical nanostructures using electrohydrodynamic (HEHD) instabilities. The created structures are readily fine-tuned, which is an important requirement for optimizing SERS to obtain the highest enhancements. HEHD pattern formation enables the fabrication of multiscale 3D structured arrays as SERS-active platforms. Importantly, each of the HEHD-patterned individual structural units yield a considerable SERS enhancement. This enables each single unit to function as an isolated sensor. Each of the formed structures can be effectively tuned and tailored to provide high SERS enhancement, while arising from different HEHD morphologies. The HEHD fabrication of sub-micrometer architectures is straightforward and robust, providing an elegant route for high-throughput biological and chemical sensing. The superior detection properties and the ability to fabricate SERS substrates on the miniaturized scale, will facilitate the development of advanced and novel opto-fluidic devices, such as portable detection systems, and will offer numerous applications in biomedical diagnostics, forensics, ecological warfare and homeland security.

Keywords: hierarchical electrohydrodynamic patterning, medical diagnostics, point-of care devices, SERS

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Authors: Niharika Kaushal, Minni Singh

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Fruit crops are widely cultivated throughout the World, with citrus being one of the most common. Mandarins, oranges, grapefruits, lemons, and limes are among the most frequently grown varieties. Citrus cultivars are industrially processed into juice, resulting in approx. 25-40% by wt. of biomass in the form of peels and seeds, generally considered as waste. In consequence, a significant amount of this nutraceutical-enriched biomass goes to waste, which, if utilized wisely, could revolutionize the functional food industry, as this biomass possesses a wide range of bioactive compounds, mainly within the class of polyphenols and terpenoids, making them an abundant source of functional bioactive. Mandarin is a potential source of bioflavonoids with putative antioxidative properties, and its potential application for developing value-added products is obvious. In this study, ‘kinnow’ mandarin (Citrus nobilis X Citrus deliciosa) biomass was studied for its flavonoid profile. For this, dried and pulverized peels were subjected to green and sustainable extraction techniques, namely, supercritical fluid extraction carried out under conditions pressure: 330 bar, temperature: 40 ̊ C and co-solvent: 10% ethanol. The obtained extract was observed to contain 47.3±1.06 mg/ml rutin equivalents as total flavonoids. Mass spectral analysis revealed the prevalence of polymethoxyflavones (PMFs), chiefly tangeretin and nobiletin. Furthermore, the antioxidant potential was analyzed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, which was estimated to be at an IC₅₀ of 0.55μg/ml. The pre-systemic metabolism of flavonoids limits their functionality, as was observed in this study through in vitro gastrointestinal studies where nearly 50.0% of the flavonoids were degraded within 2 hours of gastric exposure. We proposed nanoencapsulation as a means to overcome this problem, and flavonoids-laden polylactic-co-glycolic acid (PLGA) nano encapsulates were bioengineered using solvent evaporation method, and these were furnished to a particle size between 200-250nm, which exhibited protection of flavonoids in the gastric environment, allowing only 20% to be released in 2h. A further step involved impregnating the nano encapsulates within alginate hydrogels which were fabricated by ionic cross-linking, which would act as delivery vehicles within the gastrointestinal (GI) tract. As a result, 100% protection was achieved from the pre-systemic release of bioflavonoids. These alginate hydrogels had key significant features, i.e., less porosity of nearly 20.0%, and Cryo-SEM (Cryo-scanning electron microscopy) images of the composite corroborate the packing ability of the alginate hydrogel. As a result of this work, it is concluded that the waste can be used to develop functional biomaterials while retaining the functionality of the bioactive itself.

Keywords: bioflavonoids, gastrointestinal, hydrogels, mandarins

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Authors: Davide Cocchi, Andrea Zucchelli, Luca Raimondi, Maria Brugo Tommaso

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The increasing attention is given to the issues of environmental pollution and climate change is exponentially stimulating the development of electrically propelled vehicles powered by renewable energy, in particular, the solar one. Given the small amount of solar energy that can be stored and subsequently transformed into propulsive energy, it is necessary to develop vehicles with high mechanical, electrical and aerodynamic efficiencies along with reduced masses. The reduction of the masses is of fundamental relevance especially for the unsprung masses, that is the assembly of those elements that do not undergo a variation of their distance from the ground (wheel, suspension system, hub, upright, braking system). Therefore, the reduction of unsprung masses is fundamental in decreasing the rolling inertia and improving the drivability, comfort, and performance of the vehicle. This principle applies even more in solar propelled vehicles, equipped with an electric motor that is connected directly to the wheel hub. In this solution, the electric motor is integrated inside the wheel. Since the electric motor is part of the unsprung masses, the development of compact and lightweight solutions is of fundamental importance. The purpose of this research is the design development and optimization of a CFRP 16 wheel hub motor for solar propulsion vehicles that can carry up to four people. In addition to trying to maximize aspects of primary importance such as mass, strength, and stiffness, other innovative constructive aspects were explored. One of the main objectives has been to achieve a high geometric packing in order to ensure a reduced lateral dimension, without reducing the power exerted by the electric motor. In the final solution, it was possible to realize a wheel hub motor assembly completely comprised inside the rim width, for a total lateral overall dimension of less than 100 mm. This result was achieved by developing an innovative connection system between the wheel and the rotor with a double purpose: centering and transmission of the driving torque. This solution with appropriate interlocking noses allows the transfer of high torques and at the same time guarantees both the centering and the necessary stiffness of the transmission system. Moreover, to avoid delamination in critical areas, evaluated by means of FEM analysis using 3D Hashin damage criteria, electrospun nanofibrous mats have been interleaved between CFRP critical layers. In order to reduce rolling resistance, the rim has been designed to withstand high inflation pressure. Laboratory tests have been performed on the rim using the Digital Image Correlation technique (DIC). The wheel has been tested for fatigue bending according to E/ECE/324 R124e.

Keywords: composite laminate, delamination, DIC, lightweight vehicle, motor hub wheel, nanofiber

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Authors: Diego F. Camacho, Cristian J. Mejia, Carlos Duque-Daza

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The recent widespread use of Unmanned Aerial Vehicles (UAVs) has fueled a renewed interest in efficiency and performance of airfoils, particularly for applications at low and moderate Reynolds numbers, typical of this kind of vehicles. Most of previous efforts in the aeronautical industry, regarding aerodynamic efficiency, had been focused on high Reynolds numbers applications, typical of commercial airliners and large size aircrafts. However, in order to increase the levels of efficiency and to boost the performance of these UAV, it is necessary to explore new alternatives in terms of airfoil design and application of drag reduction techniques. The objective of the present work is to carry out the analysis and comparison of performance levels between a standard NACA0012 profile against another one featuring a wall protuberance or surface perturbation. A computational model, based on the finite volume method, is employed to evaluate the effect of the presence of geometrical distortions on the wall. The performance evaluation is achieved in terms of variations of drag and lift coefficients for the given profile. In particular, the aerodynamic performance of the new design, i.e. the airfoil with a surface perturbation, is examined under conditions of incompressible and subsonic flow in transient state. The perturbation considered is a shaped protrusion prescribed as a small surface deformation on the top wall of the aerodynamic profile. The ultimate goal by including such a controlled smooth artificial roughness was to alter the turbulent boundary layer. It is shown in the present work that such a modification has a dramatic impact on the aerodynamic characteristics of the airfoil, and if properly adjusted, in a positive way. The computational model was implemented using the unstructured, FVM-based open source C++ platform OpenFOAM. A number of numerical experiments were carried out at Reynolds number 5x104, based on the length of the chord and the free-stream velocity, and angles of attack 6° and 12°. A Large Eddy Simulation (LES) approach was used, together with the dynamic Smagorinsky approach as subgrid scale (SGS) model, in order to account for the effect of the small turbulent scales. The impact of the surface perturbation on the performance of the airfoil is judged in terms of changes in the drag and lift coefficients, as well as in terms of alterations of the main characteristics of the turbulent boundary layer on the upper wall. A dramatic change in the whole performance can be appreciated, including an arguably large level of lift-to-drag coefficient ratio increase for all angles and a size reduction of laminar separation bubble (LSB) for a twelve-angle-of-attack.

Keywords: CFD, LES, Lift-to-drag ratio, LSB, NACA 0012 airfoil

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Authors: Xiao-Mei Zhang, Sian-Hong Tseng, Ming-Yen Lu

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Two-dimensional (2D) transition metal dichalcogenides (TMDCs), such as MoS₂, have attracted considerable attention owing to the unique optical and electronic properties related to its 2D ultrathin atomic layer structure. MoS₂ is becoming prevalent in post-silicon digital electronics and in highly efficient optoelectronics due to its extremely low thickness and its tunable band gap (Eg = 1-2 eV). For low-power, high-performance complementary logic applications, both p- and n-type MoS₂ FETs (NFETs and PFETs) must be developed. NFETs with an electron accumulation channel can be obtained using unintentionally doped n-type MoS₂. However, the fabrication of MoS₂ FETs with complementary p-type characteristics is challenging due to the significant difficulty of injecting holes into its inversion channel. Plasma treatments with different species (including CF₄, SF₆, O₂, and CHF₃) have also been found to achieve the desired property modifications of MoS₂. In this work, we demonstrated a p-type multilayer MoS₂ enabled by selective-area doping using CHF₃ plasma treatment. Compared with single layer MoS₂, multilayer MoS₂ can carry a higher drive current due to its lower bandgap and multiple conduction channels. Moreover, it has three times the density of states at its minimum conduction band. Large-area growth of MoS₂ films on 300 nm thick SiO₂/Si substrate is carried out by thermal decomposition of ammonium tetrathiomolybdate, (NH₄)₂MoS₄, in a tube furnace. A two-step annealing process is conducted to synthesize MoS₂ films. For the first step, the temperature is set to 280 °C for 30 min in an N₂ rich environment at 1.8 Torr. This is done to transform (NH₄)₂MoS₄ into MoS₃. To further reduce MoS₃ into MoS₂, the second step of annealing is performed. For the second step, the temperature is set to 750 °C for 30 min in a reducing atmosphere consisting of 90% Ar and 10% H₂ at 1.8 Torr. The grown MoS₂ films are subjected to out-of-plane doping by CHF₃ plasma treatment using a Dry-etching system (ULVAC original NLD-570). The radiofrequency power of this dry-etching system is set to 100 W and the pressure is set to 7.5 mTorr. The final thickness of the treated samples is obtained by etching for 30 s. Back-gated MoS₂ PFETs were presented with an on/off current ratio in the order of 10³ and a field-effect mobility of 65.2 cm²V⁻¹s⁻¹. The MoS₂ PFETs photodetector exhibited ultraviolet (UV) photodetection capability with a rapid response time of 37 ms and exhibited modulation of the generated photocurrent by back-gate voltage. This work suggests the potential application of the mild plasma-doped p-type multilayer MoS₂ in UV photodetectors for environmental monitoring, human health monitoring, and biological analysis.

Keywords: photodetection, p-type doping, multilayers, MoS₂

Procedia PDF Downloads 104

Authors: Evgenia Papathanasiou, Georgios Kokkotis, Georgios Axiaris, Theodoros Argyropoulos, Nikos Viazis, Olga Giouleme, Konstantinos Gkoumas, Αnthia Gatopoulou, Αggelos Theodoulou, Georgios Theocharis, Αngeliki Theodoropoulou, Μaria Κalogirou, Pantelis Karatzas, Κonstantinos Κatsanos, Theodora Kafetzi, Κonstantinos Κarmiris, Αnastasia Κourikou, Ιoannis E Κoutroubakis, Christos Liatsos, Gerassimos J. Mantzaris, Νicoletta Μathou, Georgia Bellou, George Michalopoulos Αikaterini Μantaka, Penelope Nikolaou, Μichael Oikonomou, Dimitrios Polymeros, George Papatheodoridis, Εvdoxia Stergiou, Κonstantinos Soufleris, Εpameinondas Skouloudis, Μaria Tzouvala, Georgia Tsiolakidou, Εftychia Tsironi, Styliani Tsafaraki, Kalliopi Foteinogiannopoulou, Konstantina Chalakatevaki, Αngeliki Christidou, Dimitrios K. Christodoulou, Giorgos Bamias, Spyridon Michopoulos, Εvanthia Zampeli

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Background: Inflammatory bowel disease (IBD) commonly affects female patients of reproductive age, making the interaction between fertility, pregnancy and IBD an important issue in disease management. The effect of disease activity on the outcome of pregnancy and its impact on neonatal growth is a field of intense research. Close follow-up of pregnant IBD patients by a multidisciplinary team improves maternal and neonatal outcomes. Aim – Methods: Α national retrospective study of pregnancies in women with IBD between 2010-2020 was carried out in 22 IBD reference centers in Greece. Patient characteristics such as disease profile, type of treatment, and disease activity during gestation were analyzed in correlation to the method of delivery, pregnancy outcomes, as well as breastfeeding and offspring health. Results: Two-hundred and twenty-three pregnancies in 175 IBD patients were registered in the study. 122 with Crohn’s disease (CD). Median age during diagnosis was 25.6 years (12-44), with median disease duration of 7.4 years (0-23). One-hundred and twenty-nine patients (58%) were recorded during their first pregnancy. Early pregnancy termination was reported by 48 patients (22%). Pregnancy as a result of in vitro fertilization (IVF) occurred in 15 cases (6.7%). At the beginning of gestation, 165 patients (74%) were under treatment: 48 with anti-TNF agents (29%), 43 with azathioprine (26%), 101 with 5-aminosalicylic acid formulations (61%) and 12 with steroids (7%). We recorded 49 cases of IBD flares (22%) during pregnancy. Two-thirds of them (n=30) were in remission at the onset of the pregnancy. Almost half of them (n=22) required corticosteroid treatment. Patients with ulcerative colitis (UC) were in greater risk of disease flare during pregnancy (p<0.001). All but 3 pregnancies (99.1%) resulted in uncomplicated delivery. In 147 cases (67.1%), cesarean delivery was performed. Two late fetal deaths (0.9%) were reported, both in patients with continuously active disease since the beginning of pregnancy. After delivery, 75 patients (34%) presented with a disease flare, which was associated with active disease at the beginning of pregnancy (p <0.001). Conclusion: The majority of female, Greek IBD patients, had a favorable pregnancy outcome. Active inflammation during gestation and UC diagnosis were associated with a negative impact on pregnancy outcomes. The results of this study are in favor of the continuation of IBD treatment during pregnancy.

Keywords: pregnancy, ulcerative colitis, Crohn disease, flare

Procedia PDF Downloads 87

Authors: Natalia Caldeira De Carvalho, Tassia Batista Pessato, Luis Gustavo R. Fernandes, Ricardo L. Zollner, Flavia Maria Netto

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Protein hydrolysates are ingredients of enteral diets and hypoallergenic formulas. Enzymatic hydrolysis is the most commonly used method for reducing the antigenicity of milk protein. The antigenicity and physicochemical characteristics of the protein hydrolysates depend on the reaction parameters. Among them, pH has been pointed out as of the major importance. Hydrolysis reaction in laboratory scale is commonly carried out under controlled pH (pH-stat). However, from the industrial point of view, controlling pH during hydrolysis reaction may be infeasible. This study evaluated the impact of pH control on the physicochemical properties and antigenicity of the hydrolysates of whey proteins with Alcalase. Whey protein isolate (WPI) solutions containing 3 and 7 % protein (w/v) were hydrolyzed with Alcalase 50 and 100 U g-1 protein at 60°C for 180 min. The reactions were carried out under controlled and uncontrolled pH conditions. Hydrolyses performed under controlled pH (pH-stat) were initially adjusted and maintained at pH 8.5. Hydrolyses carried out without pH control were initially adjusted to pH 8.5. Degree of hydrolysis (DH) was determined by OPA method, peptides profile was evaluated by HPLC-RP, and molecular mass distribution by SDS-PAGE/Tricine. The residual α-lactalbumin (α-La) and β-lactoglobulin (β-Lg) concentrations were determined using commercial ELISA kits. The specific IgE and IgG binding capacity of hydrolysates was evaluated by ELISA technique, using polyclonal antibodies obtained by immunization of female BALB/c mice with α-La, β-Lg and BSA. In hydrolysis under uncontrolled pH, the pH dropped from 8.5 to 7.0 during the first 15 min, remaining constant throughout the process. No significant difference was observed between the DH of the hydrolysates obtained under controlled and uncontrolled pH conditions. Although all hydrolysates showed hydrophilic character and low molecular mass peptides, hydrolysates obtained with and without pH control exhibited different chromatographic profiles. Hydrolysis under uncontrolled pH released, predominantly, peptides between 3.5 and 6.5 kDa, while hydrolysis under controlled pH released peptides smaller than 3.5 kDa. Hydrolysis with Alcalase under all conditions studied decreased by 99.9% the α-La and β-Lg concentrations in the hydrolysates detected by commercial kits. In general, β-Lg concentrations detected in the hydrolysates obtained under uncontrolled pH were significantly higher (p<0.05) than those detected in hydrolysates produced with pH control. The anti-α-La and anti-β-Lg IgE and IgG responses to all hydrolysates decreased significantly compared to WPI. Levels of specific IgE and IgG to the hydrolysates were below 25 and 12 ng ml-1, respectively. Despite the differences in peptide composition and α-La and β-Lg concentrations, no significant difference was found between IgE and IgG binding capacity of hydrolysates obtained with or without pH control. These results highlight the impact of pH on the hydrolysates characteristics and their concentrations of antigenic protein. Divergence between the antigen detection by commercial ELISA kits and specific IgE and IgG binding response was found in this study. This result shows that lower protein detection does not imply in lower protein antigenicity. Thus, the use of commercial kits for allergen contamination analysis should be cautious.

Keywords: allergy, enzymatic hydrolysis, milk protein, pH conditions, physicochemical characteristics

Procedia PDF Downloads 302

Authors: A. S. Vidyashree, C. M. Kalleshwaraswamy, R. Asokan, H. M. Mahadevaswamy

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Termites are very vital ecological thespians in tropical ecosystem, having been designated as “ecosystem engineers”, due to their significant role in providing soil ecosystem services. Despite their importance, our understanding of a number of their basic biological processes in termites is extremely limited. Developing a better understanding of termite biology is closely dependent upon consistent species identification. At present, identification of termites is relied on soldier castes. But for many species, soldier caste is not reported, that creates confusion in identification. The use of molecular markers may be helpful in estimating phylogenetic relatedness between the termite species and estimating genetic differentiation among local populations within each species. To understand this, termites samples were collected from various places of Western Ghats covering four states namely Karnataka, Kerala, Tamil Nadu, Maharashtra during 2013-15. Termite samples were identified based on their morphological characteristics, molecular characteristics, or both. Survey on the termite fauna in Karnataka, Kerala, Maharashtra and Tamil Nadu indicated the presence of a 16 species belongs to 4 subfamilies under two families viz., Rhinotermitidae and Termitidae. Termititidae was the dominant family which was belonging to 4 genera and four subfamilies viz., Macrotermitinae, Amitermitinae, Nasutitermitinae and Termitinae. Amitermitinae had three species namely, Microcerotermes fletcheri, M. pakistanicus and Speculitermes sinhalensis. Macrotermitinae had the highest number of species belonging two genera, namely Microtermes and Odontotermes. Microtermes genus was with only one species i.e., Microtermes obesi. The genus Odontotermes was represented by the highest number of species (07), namely, O. obesus was the dominant (41 per cent) and the most widely distributed species in Karnataka, Karala, Maharashtra and Tamil nadu followed by O. feae (19 per cent), O.assmuthi (11 per cent) and others like O. bellahunisensis O. horni O. redemanni, O. yadevi. Nasutitermitinae was represented by two genera namely Nasutitermes anamalaiensis and Trinervitermes biformis. Termitinae subfamily was represented by Labiocapritermes distortus. Rhinotermitidae was represented by single subfamily Heterotermetinae. In Heterotermetinae, two species namely Heterotermes balwanthi and H. malabaricus were recorded. Genetic relationship among termites collected from various locations of Western Ghats of India was characterized based on mitochondrial DNA sequences (12S, 16S, and COII). Sequence analysis and divergence among the species was assessed. These results suggest that the use of both molecular and morphological approaches is crucial in ensuring accurate species identification. Efforts were made to understand their evolution and to address the ambiguities in morphological taxonomy. The implication of the study in revising the taxonomy of Indian termites, their characterization and molecular comparisons between the sequences are discussed.

Keywords: isoptera, mitochondrial DNA sequences, rhinotermitidae, termitidae, Western ghats

Procedia PDF Downloads 266

Authors: Fruzsina Luca Kézér, Viktor Jurkovich, Ottó Szenci, János Tőzsér, Levente Kovács

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Positive interaction between people and animals could have a favorable effect on the welfare and production by reducing stress levels. However, to the repeated contact with humans (e.g. farm staff, veterinarians or herdsmen), animals may respond with escape behavior or avoidance, which both have negative effects on the ease of handling, welfare and may lead to the expression of aggressive behaviors. Rough or aversive handling can impair health and the function of the cardiac autonomic activity due to fear and stress, which also can be determined by certain parameters of heart rate variability (HRV). Although the essential relationships between fear from humans and basal tone of the autonomic nervous system were described by the authors previously, several questions remained unclear in terms of the associations between different coping strategies (behavioral syndromes) of the animals and physiological responsiveness to humans. The main goal of this study was to find out whether human behavior and emotions to the animals have an impact on cardiac function and behavior of animals with different coping styles in response situations. Therefore, in the present study, special (fear, approaching, restraint, novel arena, novel object) tests were performed on healthy, 2-year old heifers (n = 104) differing in coping styles [reactive (passive) vs. proactive (active) coping]. Animals were categorized as reactive or proactive based on the following tests: 1) aggressive behavior at the feeding bunk, 2) avoidance from an approaching person, 3) immobility, and 4) daily activity (number of posture changes). Heart rate, the high frequency (HF) component of HRV as a measure of vagal activity and the ratio between the low frequency (LF) and HF components (LF/HF ratio) as a parameter of sympathetic nervous system activity were calculated for all individual during lying posture (baseline) and for response situations in novel object, novel arena, and unfamiliar person tests (both for 5 min), respectively. The differences between baseline and response were compared between groups. Higher sympathetic (higher heart rates and LF/HF ratios) and lower parasympathetic activity (lower HF) was found for proactive animals in response situations than for reactive (passive) animals either during the novel object, the novel arena and the unfamiliar person test. It suggests that animals with different behavioral traits differ in their immediate autonomic adaptation to novelty and people. Based on our preliminary results, it seems, that the analysis of HRV can help to understand the physiological manifestation of responsiveness to novelty and human presence in dairy cattle with different behavioral syndromes.

Keywords: behavioral syndromes, human-cattle interaction, novel arena test, physiological responsiveness, proactive coping, reactive coping

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Authors: Nidhi Malshe

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Background: Currently, there are approximately 1.3 billion individuals worldwide living with significant disabilities, which accounts for 16% of the global population—about 1 in 6 people. As the global population continues to grow, so does the number of people experiencing disabilities. Traffic accidents alone contribute to millions of injuries and disabilities each year, particularly among young people. Additionally, as life expectancy rises, more individuals are likely to experience disabilities in their later years. 27.0% of Canadians aged 15 and over, or 8 million people, had at least one disability in 2022. This represents an increase of 4.7 percentage points from 2017. A person with a disability earns 21.4% less on average as compared to a person without a disability. Using innovative and inclusive methods for accommodations, disability management, and employment, we can progress towards inclusive workplaces and potential income parity for this equity-seeking population. Objective: This study embraces innovative and inclusive approaches to disability management, thereby unlocking the advantages associated with a) fostering equal opportunities for all individuals, b) facilitating streamlined accommodations and making it easier for companies to accommodate people with disabilities, c) harnessing diverse perspectives to drive innovation and enhance overall productivity. Methodology: Literature review, assessments of specific needs and requirements in the workplace. a) Encourage the ability to think out of the box for potential workplace accommodations based on the specific needs of individuals. e.g., propose prolonged integration post disability. b) Perform a cost-benefit analysis of early interventions of return to work vs. duration on disability. c) Expand the scope of vocational assessment/retraining – e.g., retraining a person with permanent physical impairment to become a video game coder. d) Leverage the use of technology while planning to return to work e.g., speech-to-text software for persons with voice impairments. Hypothesized Results: Prolonged progression of return to work increases the potential for sustainable and productive employment. Co-developing a person-centric accommodation plan based on reported functional abilities and applying pioneering methods for extending accommodations to prevent secondary disabilities. Facilitate a sense of belonging by providing employees with benefits and initiatives that honor their unique contributions. Engage individuals with disabilities as active members of the planning committee to ensure the development of innovative and inclusive accommodations that address the needs of all. Conclusion: The global pandemic underscored the need for creativity in our daily routine. It is imperative to integrate the lessons learned from the pandemic, enhance them within employment, and return to work processes. These learnings can also be used to develop creative, distinct methods to ensure equal opportunities for everyone.

Keywords: disbaility management, diversity, inclusion, innovation

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Authors: Malek Miladi, Miguel Vegara, Maria Perez-Infantes, Khaled Mohamed Ramadan, Antonio Ruiz-Canales, Damaris Nunez-Gomez

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Coffee is the second consumed commodity worldwide, yet it also generates colossal waste. Proper management of coffee waste is proposed by converting them into products with higher added value to achieve sustainability of the economic and ecological footprint and protect the environment. Based on this, a study looking at the recovery of coffee waste is becoming more relevant in recent decades. Spent coffee grounds (SCG's) resulted from brewing coffee represents the major waste produced among all coffee industry. The fact that SCGs has no economic value be abundant in nature and industry, do not compete with agriculture and especially its high oil content (between 7-15% from its total dry matter weight depending on the coffee varieties, Arabica or Robusta), encourages its use as a sustainable feedstock for bio-oil production. The bio-oil extraction is a crucial step towards biodiesel production by the transesterification process. However, conventional methods used for oil extraction are not recommended due to their high consumption of energy, time, and generation of toxic volatile organic solvents. Thus, finding a sustainable, economical, and efficient extraction technique is crucial to scale up the process and to ensure more environment-friendly production. Under this perspective, the aim of this work was the statistical study to know an efficient strategy for oil extraction by n-hexane using indirect sonication. The coffee waste mixed Arabica and Robusta, which was used in this work. The temperature effect, sonication time, and solvent-to-solid ratio on the oil yield were statistically investigated as dependent variables by Central Composite Rotatable Design (CCRD) 23. The results were analyzed using STATISTICA 7 StatSoft software. The CCRD showed the significance of all the variables tested (P < 0.05) on the process output. The validation of the model by analysis of variance (ANOVA) showed good adjustment for the results obtained for a 95% confidence interval, and also, the predicted values graph vs. experimental values confirmed the satisfactory correlation between the model results. Besides, the identification of the optimum experimental conditions was based on the study of the surface response graphs (2-D and 3-D) and the critical statistical values. Based on the CCDR results, 29 ºC, 56.6 min, and solvent-to-solid ratio 16 were the better experimental conditions defined statistically for coffee waste oil extraction using n-hexane as solvent. In these conditions, the oil yield was >9% in all cases. The results confirmed the efficiency of using an ultrasound bath in extracting oil as a more economical, green, and efficient way when compared to the Soxhlet method.

Keywords: coffee waste, optimization, oil yield, statistical planning

Procedia PDF Downloads 119

Authors: Krishmali N. Ekanayake, Brendan J. Holland, Colin J. Barrow, Rick Wood

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Globally, more than one-third of all food produced is lost or wasted, equating to 1.3 billion tonnes per year. Around 31.2 million tonnes of food waste are generated across the production, supply, and consumption chain in Australia. Generally, the food waste management processes adopt environmental-friendly and more sustainable approaches such as composting, anerobic digestion and energy implemented technologies. However, unavoidable, and non-recyclable food waste ends up as landfilling and incineration that involve many undesirable impacts and challenges on the environment. A biorefinery approach contributes to a waste-minimising circular economy by converting food and other organic biomass waste into valuable outputs, including feeds, nutrition, fertilisers, and biomaterials. As a solution, Green Eco Technologies has developed a food waste treatment process using WasteMaster system. The system uses charged oxygen and moderate temperatures to convert food waste, without bacteria, additives, or water, into a virtually odour-free, much reduced quantity of reusable residual material. In the context of a biorefinery, the WasteMaster dries and mills food waste into a form suitable for storage or downstream extraction/separation/concentration to create products. The focus of the study is to determine the nutritional composition of WasteMaster processed residue to potential develop aquafeed ingredients. The global aquafeed industry is projected to reach a high value market in future, which has shown high demand for the aquafeed products. Therefore, food waste can be utilized for aquaculture feed development by reducing landfill. This framework will lessen the requirement of raw crops cultivation for aquafeed development and reduce the aquaculture footprint. In the present study, the nutritional elements of processed residue are consistent with the input food waste type, which has shown that the WasteMaster is not affecting the expected nutritional distribution. The macronutrient retention values of protein, lipid, and nitrogen free extract (NFE) are detected >85%, >80%, and >95% respectively. The sensitive food components including omega 3 and omega 6 fatty acids, amino acids, and phenolic compounds have been found intact in each residue material. Preliminary analysis suggests a price comparability with current aquafeed ingredient cost making the economic feasibility. The results suggest high potentiality of aquafeed development as 5 to 10% of the ingredients to replace/partially substitute other less sustainable ingredients across biorefinery setting. Our aim is to improve the sustainability of aquaculture and reduce the environmental impacts of food waste.

Keywords: biorefinery, ffood waste residue, input, wasteMaster

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Authors: Amenah Dhannoon, Ciaran Martin Hurley, Laura Wrafter, Podraic J. Regan

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Introduction: The COVID-19 pandemic continues to present unprecedented challenges for healthcare systems. This has resulted in the pragmatic shift in the practice of plastic surgery units worldwide. During this period, many units reported a significant fall in urgent melanoma referrals, leading to patients presenting with advanced disease requiring more extensive surgery and inferior outcomes. Our objective was to evaluate our unit's experience with both non-invasive and invasive melanoma during the COVID-19 pandemic and characterize our experience and contrast it to that experienced by our neighbors in the UK, mainland Europe and North America. Methods: a retrospective chart review was performed on all patients diagnosed with invasive and non-invasive cutaneous melanoma between March to December of 2019 (control) compared to 2020 (COVID-19 pandemic) in a single plastic surgery unit in Ireland. Patient demographics, referral source, surgical procedures, tumour characteristics, radiological findings, oncological therapies and follow-up were recorded. All data were anonymized and stored in Microsoft Excel. Results: A total of 589 patients were included in the study. Of these, 314 (53%) with invasive melanoma, compared to 275 (47%) with the non-invasive disease. Overall, more patients were diagnosed with both invasive and non-invasive melanoma in 2020 than in 2019 (p<0.05). However, significantly longer waiting times in 2020 (64 days) compared to 2019 (28 days) (p<0.05), with the majority of the referral being from GP in 2019 (83%) compared to 61% in 2020. Positive sentinel lymph node were higher in 2019 at 56% (n=28) compared to 24% (n=22) in 2020. There was no statistically significant difference in the tutor characteristics or metastasis status. Discussion: While other countries have noticed a fall in the melanoma diagnosis. Our units experienced a higher number of disease diagnoses. This can be due to multiple reasons. In Ireland, the government reached an early agreement with the private sector to continue elective surgery on an urgent basis in private hospitals. This allowed access to local anesthetic procedures and local skin cancer cases were triaged to non-COVID-19 provider centers. Our unit also adapted a fast, effective and minimal patient contact strategy for triaging skin cancer based on telemedicine. Thirdly, a skin cancer nurse specialist maintained patient follow-ups and triaging a dedicated email service. Finally, our plastic surgery service continued to maintain a virtual complex skin cancer multidisciplinary team meeting during the pandemic, ensuring local clinical governance has adhered to each clinical case. Conclusion: Our study highlights that with the prompt efficient restructuring of services, we could reserve successful management of skin cancer even in the most devastating times. It is important to reflect on the success during the pandemic and emphasize the importance of preparation for a potentially difficult future

Keywords: malignant melanoma, skin cancer, COVID-19, triage

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Authors: Kyoungrean Kim

Abstract:

Importance of deep-sea mineral resources is dramatically increasing due to the depletion of land mineral resources corresponding to increasing human’s economic activities. Korea has acquired exclusive exploration licenses at four areas which are the Clarion-Clipperton Fracture Zone in the Pacific Ocean (2002), Tonga (2008), Fiji (2011) and Indian Ocean (2014). The preparation for commercial mining of Nautilus minerals (Canada) and Lockheed martin minerals (USA) is expected by 2020. The London Protocol 1996 (LP) under International Maritime Organization (IMO) and International Seabed Authority (ISA) will set environmental guidelines for deep-sea mining until 2020, to protect marine environment. In this research, the applicability of washing/extraction treatment for the remediation of deep-sea mining tailings was mainly evaluated in order to present preliminary data to develop practical remediation technology in near future. Polymetallic nodule samples were collected at the Clarion-Clipperton Fracture Zone in the Pacific Ocean, then stored at room temperature. Samples were pulverized by using jaw crusher and ball mill then, classified into 3 particle sizes (> 63 µm, 63-20 µm, < 20 µm) by using vibratory sieve shakers (Analysette 3 Pro, Fritsch, Germany) with 63 µm and 20 µm sieve. Only the particle size 63-20 µm was used as the samples for investigation considering the lower limit of ore dressing process which is tens to 100 µm. Rhamnolipid and sodium alginate as biosurfactant and aluminum sulfate which are mainly used as flocculant were used as environmentally friendly additives. Samples were adjusted to 2% liquid with deionized water then mixed with various concentrations of additives. The mixture was stirred with a magnetic bar during specific reaction times and then the liquid phase was separated by a centrifugal separator (Thermo Fisher Scientific, USA) under 4,000 rpm for 1 h. The separated liquid was filtered with a syringe and acrylic-based filter (0.45 µm). The extracted heavy metals in the filtered liquid were then determined using a UV-Vis spectrometer (DR-5000, Hach, USA) and a heat block (DBR 200, Hach, USA) followed by US EPA methods (8506, 8009, 10217 and 10220). Polymetallic nodule was mainly composed of manganese (27%), iron (8%), nickel (1.4%), cupper (1.3 %), cobalt (1.3%) and molybdenum (0.04%). Based on remediation standards of various countries, Nickel (Ni), Copper (Cu), Cadmium (Cd) and Zinc (Zn) were selected as primary target materials. Throughout this research, the use of rhamnolipid was shown to be an effective approach for removing heavy metals in samples originated from manganese nodules. Sodium alginate might also be one of the effective additives for the remediation of deep-sea mining tailings such as polymetallic nodules. Compare to the use of rhamnolipid and sodium alginate, aluminum sulfate was more effective additive at short reaction time within 4 h. Based on these results, sequencing particle separation, selective extraction/washing, advanced filtration of liquid phase, water treatment without dewatering and solidification/stabilization may be considered as candidate technologies for the remediation of deep-sea mining tailings.

Keywords: deep-sea mining tailings, heavy metals, remediation, extraction, additives

Procedia PDF Downloads 155

Authors: M. Soleimani, D. Cree, C. Chafe, L. Bates

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The accidental release of petroleum products into the environment could have harmful consequences to our ecosystem. Different techniques such as mechanical separation, membrane filtration, incineration, treatment processes using enzymes and dispersants, bioremediation, and sorption process using sorbents have been applied for oil spill remediation. Most of the techniques investigated are too costly or do not have high enough efficiency. This study was conducted to determine the sorption performance of hemp byproducts (cellulosic materials) in terms of sorption capacity and kinetics for hydrophobic and hydrophilic fluids. In this study, heavy oil, light oil, diesel fuel, and water/water vapor were used as sorbate fluids. Hemp stalk in different forms, including loose material (hammer milled (HM) and shredded (Sh) with low bulk densities) and densified forms (pellet form (P) and crumbled pellets (CP)) with high bulk densities, were used as sorbents. The sorption/retention tests were conducted according to ASTM 726 standard. For a quick-purpose application of the sorbents, the sorption tests were conducted for 15 min, and for an ideal sorption capacity of the materials, the tests were carried out for 24 h. During the test, the sorbent material was exposed to the fluid by immersion, followed by filtration through a stainless-steel wire screen. Water vapor adsorption was carried out in a controlled environment chamber with the capability of controlling relative humidity (RH) and temperature. To determine the kinetics of sorption for each fluid and sorbent, the retention capacity also was determined intervalley for up to 24 h. To analyze the kinetics of sorption, pseudo-first-order, pseudo-second order and intraparticle diffusion models were employed with the objective of minimal deviation of the experimental results from the models. The results indicated that HM and Sh materials had the highest sorption capacity for the hydrophobic fluids with approximately 6 times compared to P and CP materials. For example, average retention values of heavy oil on HM and Sh was 560% and 470% of the mass of the sorbents, respectively. Whereas, the retention of heavy oil on P and CP was up to 85% of the mass of the sorbents. This lower sorption capacity for P and CP can be due to the less exposed surface area of these materials and compacted voids or capillary tubes in the structures. For water uptake application, HM and Sh resulted in at least 40% higher sorption capacity compared to those obtained for P and CP. On average, the performance of sorbate uptake from high to low was as follows: water, heavy oil, light oil, diesel fuel. The kinetic analysis indicated that the second-pseudo order model can describe the sorption process of the oil and diesel better than other models. However, the kinetics of water absorption was better described by the pseudo-first-order model. Acetylation of HM materials could improve its oil and diesel sorption to some extent. Water vapor adsorption of hemp fiber was a function of temperature and RH, and among the models studied, the modified Oswin model was the best model in describing this phenomenon.

Keywords: environment, fiber, petroleum, sorption

Procedia PDF Downloads 124

Authors: Magdalena Wilk-Kozubek, Magdalena Rowińska, Krzysztof Rola, Joanna Cybińska

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Leuco dye-based thermochromic systems are classified as intelligent materials because they exhibit thermally induced color changes. Thanks to this feature, they are mainly used as temperature sensors in many industrial sectors. For example, placing a thermochromic material on a chemical reactor may warn about exceeding the maximum permitted temperature for a chemical process. Usually two components, a color former and a developer are needed to produce a system with irreversible color change. The color former is an electron donating (proton accepting) compound such as fluoran leuco dye. The developer is an electron accepting (proton donating) compound such as organic carboxylic acid. When the developer melts, the color former - developer complex is created and the termochromic system becomes colored. Typically, the melting point of the applied developer determines the temperature at which the color change occurs. When the lactone ring of the color former is closed, then the dye is in its colorless state. The ring opening, induced by the addition of a proton, causes the dye to turn into its colored state. Since the color former and the developer are often solid, they can be incorporated into polymer films to facilitate their practical use in industry. The objective of this research was to fabricate a leuco dye-based termochromic system that will irreversibly change color after reaching the temperature of 100°C. For this purpose, benzofluoran leuco dye (as color former) and phenoxyacetic acid (as developer with a melting point of 100°C) were introduced into the polymer films during the drop casting process. The film preparation process was optimized in order to obtain thin films with appropriate properties such as transparency, flexibility and homogeneity. Among the optimized factors were the concentration of benzofluoran leuco dye and phenoxyacetic acid, the type, average molecular weight and concentration of the polymer, and the type and concentration of the surfactant. The selected films, containing benzofluoran leuco dye and phenoxyacetic acid, were combined by mild heat treatment. Structural characterization of single and combined films was carried out by FTIR spectroscopy, morphological analysis was performed by optical microscopy and SEM, phase transitions were examined by DSC, color changes were investigated by digital photography and UV-Vis spectroscopy, while emission changes were studied by photoluminescence spectroscopy. The resulting thermochromic system is colorless at room temperature, but after reaching 100°C the developer melts and it turns irreversibly pink. Therefore, it could be used as an additional sensor to warn against boiling of water in power plants using water cooling. Currently used electronic temperature indicators are prone to faults and unwanted third-party actions. The sensor constructed in this work is transparent, thanks to which it can be unnoticed by an outsider and constitute a reliable reference for the person responsible for the apparatus.

Keywords: color developer, leuco dye, thin film, thermochromism

Procedia PDF Downloads 99

Authors: Agnieszka Antończyk, MałGorzata Ochota, Wojciech Niżański, ZdzisłAw Kiełbowicz

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The goal of the cesarean section (CS) is the delivery of healthy, vigorous pups with the provision of surgical plane anesthesia, appropriate analgesia, and rapid recovery of the dam. In human medicine, spinal or epidural anesthesia is preferred for a cesarean section as associated with a lower risk of neonatal asphyxia and the need for resuscitation. Nevertheless, the specificity of veterinary patients makes the application of regional anesthesia as a sole technique impractical, thus to obtain patient compliance the general anesthesia is required. This study aimed to compare the influence of balanced (inhalant with epidural) and inhalant anesthesia on neonatal umbilical cord blood gas (UCBG) parameters and vitality (modified Apgar scoring). The bitches (31) undergoing elective CS were enrolled in this study. All females received a single dose of 0.2 mg/kg s.c. Meloxicam. Females were randomly assigned into two groups: Gr I (Isoflurane, n=16) and Gr IE (Isoflurane plus Epidural, n=15). Anesthesia was induced with propofol at 4-6 mg/kg to effect, and maintained with isoflurane in oxygen; in IE group epidural anesthesia was also done using lidocaine (3-4 mg/kg) into the lumbosacral space. CSs were performed using a standard mid-line approach. Directly after the puppy extraction, the umbilical cord was double clamped before the placenta detachment. The vessels were gently stretched between forceps to allow blood sampling. At least 100 mcl of mixed umbilical cord blood was collected into a heparinized syringe for further analysis. The modified Apgar scoring system (AS) was used to objectively score neonatal health and vitality immediately after birth (before first aid or neonatal care was instituted), at 5 and 20 min after birth. The neonates were scored as normal (AS 7-10), weak (AS 4-6), or critical (AS 0-3). During surgery, the IE group required a lower isoflurane concentration compared to the females in group I (MAC 1.05±0.2 and 1.4±0.13, respectively, p<0.01). All investigated UCBG parameters were not statistically different between groups. All pups had mild acidosis (pH 7.21±0.08 and 7.21±0.09 in Gr I and IE, respectively) with moderately elevated pCO2 (Gr I 57.18±11.48, Gr IE 58.74±15.07), HCO3- on the lower border (Gr I 22.58±3.24, Gr IE 22.83±3.6), lowered BE (Gr I -6.1±3.57, Gr IE -5.6±4.19) and mildly elevated level of lactates (Gr I 2.58±1.48, Gr IE2.53±1.03). The glucose levels were above the reference limits in both groups of puppies (74.50±25.32 in Gr I, 79.50±29.73 in Gr IE). The initial Apgar score results were similar in I and IE groups. However, the subsequent measurements of AS revealed significant differences between both groups. Puppies from the IE group received better AS scores at 5 and 20 min compared to the I group (6.86±2.23 and 8.06±2.06 vs 5.11±2.40 and 7.83±2.05, respectively). The obtained results demonstrated that administration of epidural anesthesia reduced the requirement for isoflurane in dams undergoing cesarean section and did not affect the neonatal umbilical blood gas results. Moreover, newborns from the epidural anesthesia group were scored significantly higher in AS at 5 and 20 min, indicating their better vitality and quicker improvement post-surgery.

Keywords: apgar scoring, balanced anesthesia, cesarean section, umbilical blood gas

Procedia PDF Downloads 177

Authors: Delphine Morales, Florian Lombart, Agathe Truchot, Pauline Maire, Pascale Vigneron, Antoine Galmiche, Catherine Lok, Muriel Vayssade

Abstract:

Targeted therapy molecules are used as a first-line treatment for metastatic melanoma with B-Raf mutation. Nevertheless, these molecules can cause side effects to patients and are efficient on 50 to 60 % of them. Indeed, melanoma cell sensitivity to targeted therapy molecules is dependent on tumor microenvironment (cell-cell and cell-extracellular matrix interactions). To better unravel factors modulating cell sensitivity to B-Raf inhibitor, we have developed and compared several melanoma models: from metastatic melanoma cells cultured as monolayer (2D) to a co-culture in a 3D dermal equivalent. Cell response was studied in different melanoma cell lines such as SK-MEL-28 (mutant B-Raf (V600E), sensitive to Vemurafenib), SK-MEL-3 (mutant B-Raf (V600E), resistant to Vemurafenib) and a primary culture of dermal human fibroblasts (HDFn). Assays have initially been performed in a monolayer cell culture (2D), then a second time on a 3D dermal equivalent (dermal human fibroblasts embedded in a collagen gel). All cell lines were treated with Vemurafenib (a B-Raf inhibitor) for 48 hours at various concentrations. Cell sensitivity to treatment was assessed under various aspects: Cell proliferation (cell counting, EdU incorporation, MTS assay), MAPK signaling pathway analysis (Western-Blotting), Apoptosis (TUNEL), Cytokine release (IL-6, IL-1α, HGF, TGF-β, TNF-α) upon Vemurafenib treatment (ELISA) and histology for 3D models. In 2D configuration, the inhibitory effect of Vemurafenib on cell proliferation was confirmed on SK-MEL-28 cells (IC50=0.5 µM), and not on the SK-MEL-3 cell line. No apoptotic signal was detected in SK-MEL-28-treated cells, suggesting a cytostatic effect of the Vemurafenib rather than a cytotoxic one. The inhibition of SK-MEL-28 cell proliferation upon treatment was correlated with a strong expression decrease of phosphorylated proteins involved in the MAPK pathway (ERK, MEK, and AKT/PKB). Vemurafenib (from 5 µM to 10 µM) also slowed down HDFn proliferation, whatever cell culture configuration (monolayer or 3D dermal equivalent). SK-MEL-28 cells cultured in the dermal equivalent were still sensitive to high Vemurafenib concentrations. To better characterize all cell population impacts (melanoma cells, dermal fibroblasts) on Vemurafenib efficacy, cytokine release is being studied in 2D and 3D models. We have successfully developed and validated a relevant 3D model, mimicking cutaneous metastatic melanoma and tumor microenvironment. This 3D melanoma model will become more complex by adding a third cell population, keratinocytes, allowing us to characterize the epidermis influence on the melanoma cell sensitivity to Vemurafenib. In the long run, the establishment of more relevant 3D melanoma models with patients’ cells might be useful for personalized therapy development. The authors would like to thank the Picardie region and the European Regional Development Fund (ERDF) 2014/2020 for the funding of this work and Oise committee of "La ligue contre le cancer".

Keywords: 3D human skin model, melanoma, tissue engineering, vemurafenib efficiency

Procedia PDF Downloads 304