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Commenced in January 2007
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Edition: International
Paper Count: 44333

Search results for: location based data

533 A Multipurpose Inertial Electrostatic Magnetic Confinement Fusion for Medical Isotopes Production

Authors: Yasser R. Shaban

Abstract:

A practical multipurpose device for medical isotopes production is most wanted for clinical centers and researches. Unfortunately, the major supply of these radioisotopes currently comes from aging sources, and there is a great deal of uneasiness in the domestic market. There are also many cases where the cost of certain radioisotopes is too high for their introduction on a commercial scale even though the isotopes might have great benefits for society. The medical isotopes such as radiotracers PET (Positron Emission Tomography), Technetium-99 m, and Iodine-131, Lutetium-177 by is feasible to be generated by a single unit named IEMC (Inertial Electrostatic Magnetic Confinement). The IEMC fusion vessel is the upgrading unit of the Inertial Electrostatic Confinement IEC fusion vessel. Comprehensive experimental works on IEC were carried earlier with promising results. The principle of inertial electrostatic magnetic confinement IEMC fusion is based on forcing the binary fuel ions to interact in the opposite directions in ions cyclotrons orbits with different kinetic energies in order to have equal compression (forces) and with different ion cyclotron frequency ω in order to increase the rate of intersection. The IEMC features greater fusion volume than IEC by several orders of magnitude. The particles rate from the IEMC approach are projected to be 8.5 x 10¹¹ (p/s), ~ 0.2 microampere proton, for D/He-3 fusion reaction and 4.2 x 10¹² (n/s) for D/T fusion reaction. The projected values of particles yield (neutrons and protons) are suitable for medical isotope productions on-site by a single unit without any change in the fusion vessel but only the fuel gas. The PET radiotracers are usually produced on-site by medical ion accelerator whereas Technetium-99m (Tc-99m) is usually produced off-site from the irradiation facilities of nuclear power plants. Typically, hospitals receive molybdenum-99 isotope container; the isotope decays to Tc-99mwith half-life time 2.75 days. Even though the projected current from IEMC is lesser than the proton current from the medical ion accelerator but still the IEMC vessel is simpler, and reduced in components and power consumption which add a new value of populating the PET radiotracers in most clinical centers. On the other hand, the projected neutrons flux from the IEMC is lesser than the thermal neutron flux at the irradiation facilities of nuclear power plants, but in the IEMC case the productions of Technetium-99m is suggested to be at the resonance region of which the resonance integral cross section is two orders of magnitude higher than the thermal flux. Thus it can be said the net activity from both is evened. Besides, the particle accelerator cannot be considered a multipurpose particles production unless a significant change is made to the accelerator to change from neutrons mode to protons mode or vice versa. In conclusion, the projected fusion yield from IEMC is a straightforward since slightly change in the primer IEC and ion source is required.

Keywords: electrostatic versus magnetic confinement fusion vessel, ion source, medical isotopes productions, neutron activation

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

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

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

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

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531 A Challenge to Conserve Moklen Ethnic House: Case Study in Tubpla Village, Phang Nga Province, Southern Thailand

Authors: M. Attavanich, H. Kobayashi

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Moklen is a sub-group of ethnic minority in Thailand. In the past, they were vagabonds of the sea. Their livelihood relied on the sea but they built temporary shelters to avoid strong wind and waves during monsoon season. Recently, they have permanently settled on land along coastal area and mangrove forest in Phang Nga and Phuket Province, Southern Thailand. Moklen people have their own housing culture: the Moklen ethnic house was built from local natural materials, indicating a unique structure and design. Its wooden structure is joined by rattan ropes. The construction process is very unique because of using body-based unit of measurement for design and construction. However, there are several threats for those unique structures. One of the most important threats on Moklen ethnic house is tsunami. Especially the 2004 Indian Ocean Tsunami caused widely damage to Southern Thailand and Phang Nga province was the most affected area. In that time, Moklen villages which are located along the coastal area also affected calamitously. In order to recover the damage in affected villages, mostly new modern style houses were provided by aid agencies. This process has caused a significant impact on Moklen housing culture. Not only tsunami, but also modernization has an influence on the changing appearance of the Moklen houses and the effect of modernization has been started to experience before the tsunami. As a result, local construction knowledge is very limited nowadays because the number of elderly people in Moklen has been decreasing drastically. Last but not the least, restrictions of construction materials which are originally provided from accessible mangroves, create limitations in building a Moklen house. In particular, after the Reserved Forest Act, wood chopping without any permission has become illegal. These are some of the most important reasons for Moklen ethnic houses to disappear. Nevertheless, according to the results of field surveys done in 2013 in Phang Nga province, it is found out that some Moklen ethnic houses are still available in Tubpla Village, but only a few. Next survey in the same area in 2014 showed that number of Moklen houses in the village has been started to increase significantly. That proves that there is a high potential to conserve Moklen houses. Also the project of our research team in February 2014 contributed to continuation of Moklen ethnic house. With the cooperation of the village leader and our team, it was aimed to construct a Moklen house with the help of local participants. For the project, villagers revealed the building knowledge and techniques, and in the end, project helped community to understand the value of their houses. Also, it was a good opportunity for Moklen children to learn about their culture. In addition, NGOs recently have started to support ecotourism projects in the village. It not only helps to preserve a way of life, but also contributes to preserve indigenous knowledge and techniques of Moklen ethnic house. This kind of supporting activities are important for the conservation of Moklen ethnic houses.

Keywords: conservation, construction project, Moklen Ethnic House, 2004 Indian Ocean tsunami

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530 Li2o Loss of Lithium Niobate Nanocrystals during High-Energy Ball-Milling

Authors: Laura Kocsor, Laszlo Peter, Laszlo Kovacs, Zsolt Kis

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The aim of our research is to prepare rare-earth-doped lithium niobate (LiNbO3) nanocrystals, having only a few dopant ions in the focal point of an exciting laser beam. These samples will be used to achieve individual addressing of the dopant ions by light beams in a confocal microscope setup. One method for the preparation of nanocrystalline materials is to reduce the particle size by mechanical grinding. High-energy ball-milling was used in several works to produce nano lithium niobate. Previously, it was reported that dry high-energy ball-milling of lithium niobate in a shaker mill results in the partial reduction of the material, which leads to a balanced formation of bipolarons and polarons yielding gray color together with oxygen release and Li2O segregation on the open surfaces. In the present work we focus on preparing LiNbO3 nanocrystals by high-energy ball-milling using a Fritsch Pulverisette 7 planetary mill. Every ball-milling process was carried out in zirconia vial with zirconia balls of different sizes (from 3 mm to 0.1 mm), wet grinding with water, and the grinding time being less than an hour. Gradually decreasing the ball size to 0.1 mm, an average particle size of about 10 nm could be obtained determined by dynamic light scattering and verified by scanning electron microscopy. High-energy ball-milling resulted in sample darkening evidenced by optical absorption spectroscopy measurements indicating that the material underwent partial reduction. The unwanted lithium oxide loss decreases the Li/Nb ratio in the crystal, strongly influencing the spectroscopic properties of lithium niobate. Zirconia contamination was found in ground samples proved by energy-dispersive X-ray spectroscopy measurements; however, it cannot be explained based on the hardness properties of the materials involved in the ball-milling process. It can be understood taking into account the presence of lithium hydroxide formed the segregated lithium oxide and water during the ball-milling process, through chemically induced abrasion. The quantity of the segregated Li2O was measured by coulometric titration. During the wet milling process in the planetary mill, it was found that the lithium oxide loss increases linearly in the early phase of the milling process, then a saturation of the Li2O loss can be seen. This change goes along with the disappearance of the relatively large particles until a relatively narrow size distribution is achieved in accord with the dynamic light scattering measurements. With the 3 mm ball size and 1100 rpm rotation rate, the mean particle size achieved is 100 nm, and the total Li2O loss is about 1.2 wt.% of the original LiNbO3. Further investigations have been done to minimize the Li2O segregation during the ball-milling process. Since the Li2O loss was observed to increase with the growing total surface of the particles, the influence of ball-milling parameters on its quantity has also been studied.

Keywords: high-energy ball-milling, lithium niobate, mechanochemical reaction, nanocrystals

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529 Carbohydrate Intake and Physical Activity Levels Modify the Association between FTO Gene Variants and Obesity and Type 2 Diabetes: First Nutrigenetics Study in an Asian Indian Population

Authors: K. S. Vimal, D. Bodhini, K. Ramya, N. Lakshmipriya, R. M. Anjana, V. Sudha, J. A. Lovegrove, V. Mohan, V. Radha

Abstract:

Gene-lifestyle interaction studies have been carried out in various populations. However, to date there are no studies in an Asian Indian population. Hence, we examined whether lifestyle factors such as diet and physical activity modify the association between fat mass and obesity–associated (FTO) gene variants and obesity and type 2 diabetes (T2D) in an Asian Indian population. We studied 734 unrelated T2D and 884 normal glucose-tolerant (NGT) participants randomly selected from the Chennai Urban Rural Epidemiology Study (CURES) in Southern India. Obesity was defined according to the World Health Organization Asia Pacific Guidelines (non-obese, BMI < 25 kg/m2; obese, BMI ≥ 25 kg/m2). Six single nucleotide polymorphisms (SNPs) in the FTO gene (rs9940128, rs7193144, rs8050136, rs918031, rs1588413 and rs11076023) identified from recent genome-wide association studies for T2D were genotyped by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing. Dietary assessment was carried out using a validated food frequency questionnaire and physical activity was based upon the self-report. Interaction analyses were performed by including the interaction terms in the model. A joint likelihood ratio test of the main SNP effects and the SNP-diet/physical activity interaction effects was used in the linear regression analyses to maximize statistical power. Statistical analyses were performed using STATA version 13. There was a significant interaction between FTO SNP rs8050136 and carbohydrate energy percentage (Pinteraction=0.04) on obesity, where the ‘A’ allele carriers of the SNP rs8050136 had 2.46 times higher risk of obesity than those with ‘CC’ genotype (P=3.0x10-5) among individuals in the highest tertile of carbohydrate energy percentage. Furthermore, among those who had lower levels of physical activity, the ‘A’ allele carriers of the SNP rs8050136 had 1.89 times higher risk of obesity than those with ‘CC’ genotype (P=4.0x10-5). We also found a borderline interaction between SNP rs11076023 and carbohydrate energy percentage (Pinteraction=0.08) on T2D, where the ‘A’ allele carriers in the highest tertile of carbohydrate energy percentage, had 1.57 times higher risk of T2D than those with ‘TT’ genotype (P=0.002). There was also a significant interaction between SNP rs11076023 and physical activity (Pinteraction=0.03) on T2D. No further significant interactions between SNPs and macronutrient intake or physical activity on obesity and T2D were observed. In conclusion, this is the first study to provide evidence for a gene-diet and gene-physical activity interaction on obesity and T2D in an Asian Indian population. These findings suggest that the association between FTO gene variants and obesity and T2D is influenced by carbohydrate intake and physical activity levels. Greater understanding of how FTO gene influences obesity and T2D through dietary and exercise interventions will advance the development of behavioral intervention and personalised lifestyle strategies predicted to reduce the development of metabolic diseases in ‘A’ allele carriers of both SNPs in this Asian Indian population.

Keywords: dietary intake, FTO, obesity, physical activity, type 2 diabetes, Asian Indian.

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528 Clinical and Analytical Performance of Glial Fibrillary Acidic Protein and Ubiquitin C-Terminal Hydrolase L1 Biomarkers for Traumatic Brain Injury in the Alinity Traumatic Brain Injury Test

Authors: Raj Chandran, Saul Datwyler, Jaime Marino, Daniel West, Karla Grasso, Adam Buss, Hina Syed, Zina Al Sahouri, Jennifer Yen, Krista Caudle, Beth McQuiston

Abstract:

The Alinity i TBI test is Therapeutic Goods Administration (TGA) registered and is a panel of in vitro diagnostic chemiluminescent microparticle immunoassays for the measurement of glial fibrillary acidic protein (GFAP) and ubiquitin C-terminal hydrolase L1 (UCH-L1) in plasma and serum. The Alinity i TBI performance was evaluated in a multi-center pivotal study to demonstrate the capability to assist in determining the need for a CT scan of the head in adult subjects (age 18+) presenting with suspected mild TBI (traumatic brain injury) with a Glasgow Coma Scale score of 13 to 15. TBI has been recognized as an important cause of death and disability and is a growing public health problem. An estimated 69 million people globally experience a TBI annually1. Blood-based biomarkers such as glial fibrillary acidic protein (GFAP) and ubiquitin C-terminal hydrolase L1 (UCH-L1) have shown utility to predict acute traumatic intracranial injury on head CT scans after TBI. A pivotal study using prospectively collected archived (frozen) plasma specimens was conducted to establish the clinical performance of the TBI test on the Alinity i system. The specimens were originally collected in a prospective, multi-center clinical study. Testing of the specimens was performed at three clinical sites in the United States. Performance characteristics such as detection limits, imprecision, linearity, measuring interval, expected values, and interferences were established following Clinical and Laboratory Standards Institute (CLSI) guidance. Of the 1899 mild TBI subjects, 120 had positive head CT scan results; 116 of the 120 specimens had a positive TBI interpretation (Sensitivity 96.7%; 95% CI: 91.7%, 98.7%). Of the 1779 subjects with negative CT scan results, 713 had a negative TBI interpretation (Specificity 40.1%; 95% CI: 37.8, 42.4). The negative predictive value (NPV) of the test was 99.4% (713/717, 95% CI: 98.6%, 99.8%). The analytical measuring interval (AMI) extends from the limit of quantitation (LoQ) to the upper LoQ and is determined by the range that demonstrates acceptable performance for linearity, imprecision, and bias. The AMI is 6.1 to 42,000 pg/mL for GFAP and 26.3 to 25,000 pg/mL for UCH-L1. Overall, within-laboratory imprecision (20 day) ranged from 3.7 to 5.9% CV for GFAP and 3.0 to 6.0% CV for UCH-L1, when including lot and instrument variances. The Alinity i TBI clinical performance results demonstrated high sensitivity and high NPV, supporting the utility to assist in determining the need for a head CT scan in subjects presenting to the emergency department with suspected mild TBI. The GFAP and UCH-L1 assays show robust analytical performance across a broad concentration range of GFAP and UCH-L1 and may serve as a valuable tool to help evaluate TBI patients across the spectrum of mild to severe injury.

Keywords: biomarker, diagnostic, neurology, TBI

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527 Determination of Energy and Nutrients Composition of Potential Ready-to-Use Therapeutic Food Formulated from Locally Available Resources

Authors: Amina Sa'id Muhammad, Asmau Ishaq Alhassan, Beba Raymond, Fatima Bello

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Severe acute malnutrition (SAM) remains a major killer of children under five years of age. Nigeria has the second highest burden of stunted children in the world, with a national prevalence rate of 32 percent of children under five. An estimated 2 million children in Nigeria suffer from severe acute malnutrition (SAM), and 3.9% of children in northwest Nigeria suffer from SAM, which is significantly higher than the national average of 2.1%. Community-Based Management of Acute Malnutrition (CMAM) has proven to be an effective intervention in the treatment of SAM in children using Ready-to-Use Therapeutic Food (RUTF). Ready-to-use therapeutic food (RUTF) is a key component for the treatment of Severe Acute Malnutrition. It contains all the energy and nutrients required for rapid catch-up growth and used particularly in the treatment of children over 6 months of age with SAM without medical complications. However, almost all RUTFs are currently imported to Nigeria from other countries. Shortages of RUTF due to logistics (shipping costs, delays, donor fatigue etc) and funding issues present a threat to the achievement of the 2030 World Health Assembly (WHA) targets for reducing malnutrition in addition to 2030 SDGs 2 (Zero Hunger), 3 (Good Health and Wellbeing), 12 (Responsible Consumption and Production), and 17 (Partnerships for the Goals), thus undermining its effectiveness in combating malnutrition On the other hand, the availability of human and material resources that will aid local production of RUTF presents an opportunity to fill in the gap in regular RUTF supply. About one thousand Nigerian children die of malnutrition-related causes every day, reaching a total of 361,000 each year. Owing to the high burden of malnutrition in Nigeria, the local production of RUTF is a logical step, that will ensure increased availability, acceptability, access, and efficiency in supply, and at lower costs. Objective(s): The objectives of this study were therefore, to formulate RUTF from locally available resources and to determine its energy and nutrients composition, incommensurate with the standard/commercial RUTF. Methods: Three samples of RUTF were formulated using locally available resources (soya beans, wheat, rice, baobab, brown-sugar, date palm and soya oil); which were subjected to various analysis to determine their energy/proximate composition, vitamin and mineral contents and organoleptic properties were also determined using sensory evaluation. Results: The energy values of the three samples of locally produced RUTF were found to be in conformity with WHO recommendation of ≥ 500 kcal per 100g. The energy values of the three RUTF samples produced in the current study were found to be 563.08, 503.67 and 528.98 kcal respectively. Sample A, B and C had protein content of 13.56% 16.71% and 14.62% respectively, which were higher than that of commercial RUTF (10.9%). Conclusions/recommendations: The locally formulated RUTF samples had energy value of more than 500 kcal per 100g; with an appreciable amount of macro and micro nutrients. The appearance, taste, flavor and general acceptability of the formulated RUTF samples were also commendable.

Keywords: energy, malnutrition, nutrients, RUTF

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526 Caged Compounds as Light-Dependent Initiators for Enzyme Catalysis Reactions

Authors: Emma Castiglioni, Nigel Scrutton, Derren Heyes, Alistair Fielding

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By using light as trigger, it is possible to study many biological processes, such as the activity of genes, proteins, and other molecules, with precise spatiotemporal control. Caged compounds, where biologically active molecules are generated from an inert precursor upon laser photolysis, offer the potential to initiate such biological reactions with high temporal resolution. As light acts as the trigger for cleaving the protecting group, the ‘caging’ technique provides a number of advantages as it can be intracellular, rapid and controlled in a quantitative manner. We are developing caging strategies to study the catalytic cycle of a number of enzyme systems, such as nitric oxide synthase and ethanolamine ammonia lyase. These include the use of caged substrates, caged electrons and the possibility of caging the enzyme itself. In addition, we are developing a novel freeze-quench instrument to study these reactions, which combines rapid mixing and flashing capabilities. Reaction intermediates will be trapped at low temperatures and will be analysed by using electron paramagnetic resonance (EPR) spectroscopy to identify the involvement of any radical species during catalysis. EPR techniques typically require relatively long measurement times and very often, low temperatures to fully characterise these short-lived species. Therefore, common rapid mixing techniques, such as stopped-flow or quench-flow are not directly suitable. However, the combination of rapid freeze-quench (RFQ) followed by EPR analysis provides the ideal approach to kinetically trap and spectroscopically characterise these transient radical species. In a typical RFQ experiment, two reagent solutions are delivered to the mixer via two syringes driven by a pneumatic actuator or stepper motor. The new mixed solution is then sprayed into a cryogenic liquid or surface, and the frozen sample is then collected and packed into an EPR tube for analysis. The earliest RFQ instrument consisted of a hydraulic ram unit as a drive unit with direct spraying of the sample into a cryogenic liquid (nitrogen, isopentane or petroleum). Improvements to the RFQ technique have arisen from the design of new mixers in order to reduce both the volume and the mixing time. In addition, the cryogenic isopentane bath has been coupled to a filtering system or replaced by spraying the solution onto a surface that is frozen via thermal conductivity with a cryogenic liquid. In our work, we are developing a novel RFQ instrument which combines the freeze-quench technology with flashing capabilities to enable the studies of both thermally-activated and light-activated biological reactions. This instrument also uses a new rotating plate design based on magnetic couplings and removes the need for mechanical motorised rotation, which can otherwise be problematic at cryogenic temperatures.

Keywords: caged compounds, freeze-quench apparatus, photolysis, radicals

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525 How Whatsappization of the Chatbot Affects User Satisfaction, Trust, and Acceptance in a Drive-Sharing Task

Authors: Nirit Gavish, Rotem Halutz, Liad Neta

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Nowadays, chatbots are gaining more and more attention due to the advent of large language models. One of the important considerations in chatbot design is how to create an interface to achieve high user satisfaction, trust, and acceptance. Since WhatsApp conversations sometimes substitute for face-to-face communication, we studied whether WhatsAppization of the chatbot -making the conversation resemble a WhatsApp conversation more- will improve user satisfaction, trust, and acceptance, or whether the opposite will occur due to the Uncanny Valley (UV) effect. The task was a drive-sharing task, in which participants communicated with a textual chatbot via WhatsApp and could decide whether to participate in a ride to college with a driver suggested by the chatbot. WhatsAppization of the chatbot was done in two ways: By a dialog-style conversation (Dialog versus No Dialog), and by adding WhatsApp indicators – “Last Seen”, “Connected”, “Read Receipts”, and “Typing…” (Indicators versus No Indicators). Our 120 participants were randomly assigned to one of the four 2 by 2 design groups, with 30 participants in each. They interacted with the WhatsApp chatbot and then filled out a questionnaire. The results demonstrated that, as expected from the manipulation, the interaction with the chatbot was longer for the dialog condition compared to the no dialog. This extra interaction, however, did not lead to higher acceptance -quite the opposite, since participants in the dialog condition were less willing to implement the decision made at the end of the conversation with the chatbot and continue the interaction with the driver they chose. The results are even more striking when considering the Indicators condition. Both for the satisfaction measures and the trust measures, participants’ ratings were lower in the Indicators condition compared to the No Indicators. Participants in the Indicators condition felt that the ride search process was harder to operate, and slower (even though the actual interaction time was similar). They were less convinced that the chatbot suggested real trips and they trusted the person offering the ride and referred to them by the chatbot less. These effects were more evident for participants who preferred to share their rides using WhatsApp compared to participants who preferred chatbots for that purpose. Considering our findings, we can say that the WhatsAppization of the chatbot was detrimental. This is true for the both chatbot WhatsAppization methods – by making the conversation more a dialog and adding WhatsApp indicators. For the chosen drive-sharing task, the results were, in addition to lower satisfaction, less trust in the chatbot’s suggestion and even in the driver suggested by the chatbot, and lower willingness to actually undertake the suggested ride. In addition, it seems that the most problematic WhatsAppization method was using WhatsApp’s indicators during the interaction with the chatbot. The current study suggests that a conversation with an artificial agent should also not imitate a WhatsApp conversation very closely. With the proliferation of WhatsApp use, the emotional and social aspect of face-to face commination are moving to WhatsApp communication. Based on the current study’s findings, it is possible that the UV effect also occurs in WhatsAppization, and not only in humanization, of the chatbot, with a similar feeling of eeriness, and is more pronounced for people who prefer to use WhatsApp over chatbots. The current research can serve as a starting point to study the very interesting and important topic of chatbots WhatsAppization. More methods of WhatsAppization and other tasks could be the focus of further studies.

Keywords: chatbot, WhatsApp, humanization, Uncanny Valley, drive sharing

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524 We Have Never Seen a Dermatologist. Reaching the Unreachable Through Teledermatology

Authors: Innocent Atuhe, Babra Nalwadda, Grace Mulyowa Kitunzi, Annabella Haninka Ejiri

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Background: Atopic Dermatitis (AD) is one of the most prevalent and growing chronic inflammatory skin diseases in African prisons. AD care is limited in African due to lack of information about the disease amongst primary care workers, limited access to dermatologists, lack of proper training of healthcare workers, and shortage of appropriate treatments. We designed and implemented the Prisons Telederma project based on the recommendations of the International Society of Atopic Dermatitis. Our overall goal was to increase access to dermatologist-led care for prisoners with AD through teledermatology in Uganda. We aimed to; i) to increase awareness and understanding of teledermatology among prison health workers; and ii) to improve treatment outcomes of prisoners with atopic dermatitis through increased access to and utilization of consultant dermatologists through teledermatology in Uganda prisons: Approach: We used Store-and-forward Teledermatology (SAF-TD) to increase access to dermatologist-led care for prisoners and prisons staff with AD. We conducted a five days training for prison health workers using an adapted WHO training guide on recognizing neglected tropical diseases through changes on the skin together with an adapted American Academy of Dermatology (AAD) Childhood AD Basic Dermatology Curriculum designed to help trainees develop a clinical approach to the evaluation and initial management of patients with AD. This training was followed by blended e-learning, webinars facilitated by consultant Dermatologists with local knowledge of medication and local practices, apps adjusted for pigmented skin, WhatsApp group discussions, and sharing pigmented skin AD pictures and treatment via zoom meetings. We hired a team of Ugandan Senior Consultant dermatologists to draft an iconographic atlas of the main dermatoses in pigmented African skin and shared this atlas with prison health staff for use as a job aid. We had planned to use MySkinSelfie mobile phone application to take and share skin pictures of prisoners with AD with Consultant Dermatologists, who would review the pictures and prescribe appropriate treatment. Unfortunately, the National Health Service withdrew the app from the market due to technical issues. We monitored and evaluated treatment outcomes using the Patient Oriented Eczema Measure (POEM) tool. We held four advocacy meetings to persuade relevant stakeholders to increase supplies and availability of first-line AD treatments such as emollients in prison health facilities. Results: Draft iconographic atlas of the main dermatoses in pigmented African skin Increased proportion of prison health staff with adequate knowledge of AD and teledermatology from 20% to 80% Increased proportion of prisoners with AD reporting improvement in disease severity (POEM scores) from 25% to 35% in one year. Increased proportion of prisoners with AD seen by consultant dermatologist through teledermatology from 0% to 20% in one year. Increased the availability of AD recommended treatments in prisons health facilities from 5% to 10% in one year

Keywords: teledermatology, prisoners, reaching, un-reachable

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523 An Integrated Approach to Cultural Heritage Management in the Indian Context

Authors: T. Lakshmi Priya

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With the widening definition of heritage, the challenges of heritage management has become more complex . Today heritage not only includes significant monuments but comprises historic areas / sites, historic cities, cultural landscapes, and living heritage sites. There is a need for a comprehensive understanding of the values associated with these heritage resources, which will enable their protection and management. These diverse cultural resources are managed by multiple agencies having their own way of operating in the heritage sites. An Integrated approach to management of these cultural resources ensures its sustainability for the future generation. This paper outlines the importance of an integrated approach for the management and protection of complex heritage sites in India by examining four case studies. The methodology for this study is based on secondary research and primary surveys conducted during the preparation of the conservation management plansfor the various sites. The primary survey included basic documentation, inventorying, and community surveys. Red Fort located in the city of Delhi is one of the most significant forts built in 1639 by the Mughal Emperor Shahjahan. This fort is a national icon and stands testimony to the various historical events . It is on the ramparts of Red Fort that the national flag was unfurled on 15th August 1947, when India became independent, which continues even today. Management of this complex fort necessitated the need for an integrated approach, where in the needs of the official and non official stakeholders were addressed. The understanding of the inherent values and significance of this site was arrived through a systematic methodology of inventorying and mapping of information. Hampi, located in southern part of India, is a living heritage site inscribed in the World Heritage list in 1986. The site comprises of settlements, built heritage structures, traditional water systems, forest, agricultural fields and the remains of the metropolis of the 16th century Vijayanagar empire. As Hampi is a living heritage site having traditional systems of management and practices, the aim has been to include these practices in the current management so that there is continuity in belief, thought and practice. The existing national, regional and local planning instruments have been examined and the local concerns have been addressed.A comprehensive understanding of the site, achieved through an integrated model, is being translated to an action plan which safeguards the inherent values of the site. This paper also examines the case of the 20th century heritage building of National Archives of India, Delhi and protection of a 12th century Tomb of Sultan Ghari located in south Delhi. A comprehensive understanding of the site, lead to the delineation of the Archaeological Park of Sultan Ghari, in the current Master Plan for Delhi, for the protection of the tomb and the settlement around it. Through this study it is concluded that the approach of Integrated Conservation has enabled decision making that sustains the values of these complex heritage sites in Indian context.

Keywords: conservation, integrated, management, approach

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522 The Concept of Path in Original Buddhism and the Concept of Psychotherapeutic Improvement

Authors: Beth Jacobs

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The landmark movement of Western clinical psychology in the 20th century was the development of psychotherapy. The landmark movement of clinical psychology in the 21st century will be the absorption of meditation practices from Buddhist psychology. While millions of people explore meditation and related philosophy, very few people are exposed to the materials of original Buddhism on this topic, especially to the Theravadan Abhidharma. The Abhidharma is an intricate system of lists and matrixes that were used to understand and remember Buddha’s teaching. The Abhidharma delineates the first psychological system of Buddhism, how the mind works in the universe of reality and why meditation training strengthens and purifies the experience of life. Its lists outline the psychology of mental constructions, perception, emotion and cosmological causation. While the Abhidharma is technical, elaborate and complex, its essential purpose relates to the central purpose of clinical psychology: to relieve human suffering. Like Western depth psychology, the methodology rests on understanding underlying processes of consciousness and perception. What clinical psychologists might describe as therapeutic improvement, the Abhidharma delineates as a specific pathway of purified actions of consciousness. This paper discusses the concept of 'path' as presented in aspects of the Theravadan Abhidharma and relates this to current clinical psychological views of therapy outcomes and gains. The core path in Buddhism is the Eight-Fold Path, which is the fourth noble truth and the launching of activity toward liberation. The path is not composed of eight ordinal steps; it’s eight-fold and is described as opening the way, not funneling choices. The specific path in the Abhidharma is described in many steps of development of consciousness activities. The path is not something a human moves on, but something that moments of consciousness develop within. 'Cittas' are extensively described in the Abhidharma as the atomic-level unit of a raw action of consciousness touching upon an object in a field, and there are 121 types of cittas categorized. The cittas are embedded in the mental factors, which could be described as the psychological packaging elements of our experiences of consciousness. Based on these constellations of infinitesimal, linked occurrences of consciousness, citta are categorized by dimensions of purification. A path is a chain of citta developing through causes and conditions. There are no selves, no pronouns in the Abhidharma. Instead of me walking a path, this is about a person working with conditions to cultivate a stream of consciousness that is pure, immediate, direct and generous. The same effort, in very different terms, informs the work of most psychotherapies. Depth psychology seeks to release the bound, unconscious elements of mental process into the clarity of realization. Cognitive and behavioral psychologies work on breaking down automatic thought valuations and actions, changing schemas and interpersonal dynamics. Understanding how the original Buddhist concept of positive human development relates to the clinical psychological concept of therapy weaves together two brilliant systems of thought on the development of human well being.

Keywords: Abhidharma, Buddhist path, clinical psychology, psychotherapeutic outcome

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521 Reading and Writing Memories in Artificial and Human Reasoning

Authors: Ian O'Loughlin

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Memory networks aim to integrate some of the recent successes in machine learning with a dynamic memory base that can be updated and deployed in artificial reasoning tasks. These models involve training networks to identify, update, and operate over stored elements in a large memory array in order, for example, to ably perform question and answer tasks parsing real-world and simulated discourses. This family of approaches still faces numerous challenges: the performance of these network models in simulated domains remains considerably better than in open, real-world domains, wide-context cues remain elusive in parsing words and sentences, and even moderately complex sentence structures remain problematic. This innovation, employing an array of stored and updatable ‘memory’ elements over which the system operates as it parses text input and develops responses to questions, is a compelling one for at least two reasons: first, it addresses one of the difficulties that standard machine learning techniques face, by providing a way to store a large bank of facts, offering a way forward for the kinds of long-term reasoning that, for example, recurrent neural networks trained on a corpus have difficulty performing. Second, the addition of a stored long-term memory component in artificial reasoning seems psychologically plausible; human reasoning appears replete with invocations of long-term memory, and the stored but dynamic elements in the arrays of memory networks are deeply reminiscent of the way that human memory is readily and often characterized. However, this apparent psychological plausibility is belied by a recent turn in the study of human memory in cognitive science. In recent years, the very notion that there is a stored element which enables remembering, however dynamic or reconstructive it may be, has come under deep suspicion. In the wake of constructive memory studies, amnesia and impairment studies, and studies of implicit memory—as well as following considerations from the cognitive neuroscience of memory and conceptual analyses from the philosophy of mind and cognitive science—researchers are now rejecting storage and retrieval, even in principle, and instead seeking and developing models of human memory wherein plasticity and dynamics are the rule rather than the exception. In these models, storage is entirely avoided by modeling memory using a recurrent neural network designed to fit a preconceived energy function that attains zero values only for desired memory patterns, so that these patterns are the sole stable equilibrium points in the attractor network. So although the array of long-term memory elements in memory networks seem psychologically appropriate for reasoning systems, they may actually be incurring difficulties that are theoretically analogous to those that older, storage-based models of human memory have demonstrated. The kind of emergent stability found in the attractor network models more closely fits our best understanding of human long-term memory than do the memory network arrays, despite appearances to the contrary.

Keywords: artificial reasoning, human memory, machine learning, neural networks

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520 Mapping Alternative Education in Italy: The Case of Popular and Second-Chance Schools and Interventions in Lombardy

Authors: Valeria Cotza

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School drop-out is a multifactorial phenomenon that in Italy concerns all those underage students who, at different school stages (up to 16 years old) or training (up to 18 years old), manifest educational difficulties from dropping out of compulsory education without obtaining a qualification to repetition rates and absenteeism. From the 1980s to the 2000s, there was a progressive attenuation of the economic and social model towards a multifactorial reading of the phenomenon, and the European Commission noted the importance of learning about the phenomenon through approaches able to integrate large-scale quantitative surveys with qualitative analyses. It is not a matter of identifying the contextual factors affecting the phenomenon but problematising them by means of systemic and comprehensive in-depth analysis. So, a privileged point of observation and field of intervention are those schools that propose alternative models of teaching and learning to the traditional ones, such as popular and second-chance schools. Alternative schools and interventions grew in these years in Europe as well as in the US and Latin America, working in the direction of greater equity to create the conditions (often absent in conventional schools) for everyone to achieve educational goals. Against extensive Anglo-Saxon and US literature on this topic, there is yet no unambiguous definition of alternative education, especially in Europe, where second-chance education has been most studied. There is little literature on a second chance in Italy and almost none on alternative education (with the exception of method schools, to which in Italy the concept of “alternative” is linked). This research aims to fill the gap by systematically surveying the alternative interventions in the area and beginning to explore some models of popular and second-chance schools and experiences through a mixed methods approach. So, the main research objectives concern the spread of alternative education in the Lombardy region, the main characteristics of these schools and interventions, and their effectiveness in terms of students’ well-being and school results. This paper seeks to answer the first point by presenting the preliminary results of the first phase of the project dedicated to mapping. Through the Google Forms platform, a questionnaire is being distributed to all schools in Lombardy and some schools in the rest of Italy to map the presence of alternative schools and interventions and their main characteristics. The distribution is also taking place thanks to the support of the Milan Territorial and Lombardy Regional School Offices. Moreover, other social realities outside the school system (such as cooperatives and cultural associations) can be questioned. The schools and other realities to be questioned outside Lombardy will also be identified with the support of INDIRE (Istituto Nazionale per Documentazione, Innovazione e Ricerca Educativa, “National Institute for Documentation, Innovation and Educational Research”) and based on existing literature and the indicators of “Futura” Plan of the PNRR (Piano Nazionale di Ripresa e Resilienza, “National Recovery and Resilience Plan”). Mapping will be crucial and functional for the subsequent qualitative and quantitative phase, which will make use of statistical analysis and constructivist grounded theory.

Keywords: school drop-out, alternative education, popular and second-chance schools, map

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519 Integrations of the Instructional System Design for Students Learning Achievement Motives and Science Attitudes with Stem Educational Model on Stoichiometry Issue in Chemistry Classes with Different Genders

Authors: Tiptunya Duangsri, Panwilai Chomchid, Natchanok Jansawang

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This research study was to investigate of education decisions must be made which a part of it should be passed on to future generations as obligatory for all members of a chemistry class for students who will prepare themselves for a special position. The descriptions of instructional design were provided and the recent criticisms are discussed. This research study to an outline of an integrative framework for the description of information and the instructional design model give structure to negotiate a semblance of conscious understanding. The aims of this study are to describe the instructional design model for comparisons between students’ genders of their effects on STEM educational learning achievement motives to their science attitudes and logical thinking abilities with a sample size of 18 students at the 11th grade level with the cluster random sampling technique in Mahawichanukul School were designed. The chemistry learning environment was administered with the STEM education method. To build up the 5-instrument lesson instructional plan issues were instructed innovations, the 30-item Logical Thinking Test (LTT) on 5 scales, namely; Inference, Recognition of Assumptions, Deduction, Interpretation and Evaluation scales was used. Students’ responses of their perceptions with the Test Of Chemistry-Related Attitude (TOCRA) were assessed of their attitude in science toward chemistry. The validity from Index Objective Congruence value (IOC) checked by five expert specialist educator in two chemistry classroom targets in STEM education, the E1/E2 process were equaled evidence of 84.05/81.42 which results based on criteria are higher than of 80/80 standard level with the IOC from the expert educators. Comparisons between students’ learning achievement motives with STEM educational model on stoichiometry issue in chemistry classes with different genders were differentiated at evidence level of .05, significantly. Associations between students’ learning achievement motives on their posttest outcomes and logical thinking abilities, the predictive efficiency (R2) values indicate that 69% and 70% of the variances in different male and female student groups of their logical thinking abilities. The predictive efficiency (R2) values indicate that 73%; and 74% of the variances in different male and female student groups of their science attitudes toward chemistry were associated. Statistically significant on students’ perceptions of their chemistry learning classroom environment and their science attitude toward chemistry when using the MCI and TOCRA, the predictive efficiency (R2) values indicated that 72% and 74% of the variances in different male and female student groups of their chemistry classroom climate, consequently. Suggestions that supporting chemistry or science teachers from science, technology, engineering and mathematics (STEM) in addressing complex teaching and learning issues related instructional design to develop, teach, and assess traditional are important strategies with a focus on STEM education instructional method.

Keywords: development, the instructional design model, students learning achievement motives, science attitudes with STEM educational model, stoichiometry issue, chemistry classes, genders

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518 Analyzing the Effect of Socio-Political Context on Tourism: Perceptions of Young Tourists in Greece, Portugal and Israel

Authors: Shosh Shahrabani, Sharon Teitler-Regev, Helena Desivilya Syna, Fotini Voulgaris, Evangelos Tsoukatos, Vitor Ambrosio, Sandra M. Correia Loureiro

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International crises that affect tourism, such as terror attacks, political unrest, and economic crises have become more frequent, and their influence has become broader. The influence of such extreme events depends on their salience in the tourists' awareness. Hence, it is important to understand the mechanisms underlying tourists' selection of travel destinations, especially their perceptions of crisis-related events and the impact of the sociopolitical and economic context in their countries of origin. The current study examined how the socio-political and economic context in the home countries of potential young tourists affected their selection of travel destinations. The objective was to elucidate how the salience of various crises (economic and political) in the tourists' perceptions, due to their experiences at home, color their construal of destinations affected by similar hazards and influence their travel intentions. The study focused on student tourists from Israel, Greece, and Portugal. Today about a fifth of international tourism is based on young people, especially students. These countries were chosen since Greece and Portugal are in the midst of economic crises. In addition, Greece and Portugal have experienced political instability, while Israel has security-related problems (including terrorist incidents). In 2013, a total of 648 students, responded to a questionnaire that included questions concerning attitudes and risk perceptions regarding travel to destinations with various risk hazards as well as socio-demographic details. The results indicate that over half of the Israelis intend to visit Greece or Portugal. The majority of the Portuguese intend to visit Greece, while less than a third of them intend to visit Israel. About half of the Greeks intend to visit Portugal, and most of them do not intend to visit Israel. The results indicate that greater perceived importance of economic crises mitigates the intention to travel to destinations with economic crises for tourists from origin countries that are also marked by economic crises, such as Greece and Portugal. However, for tourists from Israel, a country with a relatively stable economy, issues related to the economy barely affect their intention to travel to the other two countries. The findings also suggest that Greeks and Portuguese who are highly concerned about political unrest are unlikely to select Israel as a tourist destination. In addition, strong apprehension regarding terrorism impedes the intention to travel to destinations marked by terrorist incidents, such as Israel. The current research contributes to the existing literature by highlighting the impact of travelers' personal previous experience with crisis on their risk perceptions and in turn on their intentions to travel to countries with similar risks. Therefore, in a world where such incidents are on the rise, understanding tourists' risk perceptions and behavior and the factors influencing their destination-related decisions are crucial for countries that wish to increase the numbers of incoming tourists.

Keywords: economic crises, political instability, risk perception, young tourists

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517 Resolving a Piping Vibration Problem by Installing Viscous Damper Supports

Authors: Carlos Herrera Sierralta, Husain M. Muslim, Meshal T. Alsaiari, Daniel Fischer

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Preventing piping fatigue flow induced vibration in the Oil & Gas sector demands not only the constant development of engineering design methodologies based on available software packages, but also special piping support technologies for designing safe and reliable piping systems. The vast majority of piping vibration problems in the Oil & Gas industry are provoked by the process flow characteristics which are basically intrinsically related to the fluid properties, the type of service and its different operational scenarios. In general, the corrective actions recommended for flow induced vibration in piping systems can be grouped in two major areas: those which affect the excitation mechanisms typically associated to process variables, and those which affect the response mechanism of the pipework per se, and the pipework associated steel support structure. Where possible the first option is to try to solve the flow induced problem from the excitation mechanism perspective. However, in producing facilities the approach of changing process parameters might not always be convenient as it could lead to reduction of production rates or it may require the shutdown of the system in order to perform the required piping modification. That impediment might lead to a second option, which is to modify the response of the piping system to excitation generated by the type of process flow. In principle, the action of shifting the natural frequency of the system well above the frequency inherent to the process always favours the elimination, or considerably reduces, the level of vibration experienced by the piping system. Tightening up the clearances at the supports (ideally zero gap), and adding new static supports at the system, are typical ways of increasing the natural frequency of the piping system. However, only stiffening the piping system may not be sufficient to resolve the vibration problem, and in some cases, it might not be feasible to implement it at all, as the available piping layout could create limitations on adding supports due to thermal expansion/contraction requirements. In these cases, utilization of viscous damper supports could be recommended as these devices can allow relatively large quasi-static movement of piping while providing sufficient capabilities of dissipating the vibration. Therefore, when correctly selected and installed, viscous damper supports can provide a significant effect on the response of the piping system over a wide range of frequencies. Viscous dampers cannot be used to support sustained, static loads. This paper shows over a real case example, a methodology which allows to determine the selection of the viscous damper supports via a dynamic analysis model. By implementing this methodology, it was possible to resolve the piping vibration problem throughout redesigning adequately the existing static piping supports and by adding new viscous dampers supports. This was conducted on-stream at the oil crude pipeline in question without the necessity of reducing the production of the plant. Concluding that the application of the methodology of this paper can be applied to solve similar cases in a straightforward manner.

Keywords: dynamic analysis, flow induced vibration, piping supports, turbulent flow, slug flow, viscous damper

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516 Polar Bears in Antarctica: An Analysis of Treaty Barriers

Authors: Madison Hall

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The Assisted Colonization of Polar Bears to Antarctica requires a careful analysis of treaties to understand existing legal barriers to Ursus maritimus transport and movement. An absence of land-based migration routes prevent polar bears from accessing southern polar regions on their own. This lack of access is compounded by current treaties which limit human intervention and assistance to ford these physical and legal barriers. In a time of massive planetary extinctions, Assisted Colonization posits that certain endangered species may be prime candidates for relocation to hospitable environments to which they have never previously had access. By analyzing existing treaties, this paper will examine how polar bears are limited in movement by humankind’s legal barriers. International treaties may be considered codified reflections of anthropocentric values of the best knowledge and understanding of an identified problem at a set point in time, as understood through the human lens. Even as human social values and scientific insights evolve, so too must treaties evolve which specify legal frameworks and structures impacting keystone species and related biomes. Due to costs and other myriad difficulties, only a very select number of species will be given this opportunity. While some species move into new regions and are then deemed invasive, Assisted Colonization considers that some assistance may be mandated due to the nature of humankind’s role in climate change. This moral question and ethical imperative against the backdrop of escalating climate impacts, drives the question forward; what is the potential for successfully relocating a select handful of charismatic and ecologically important life forms? Is it possible to reimagine a different, but balanced Antarctic ecosystem? Listed as a threatened species under the U.S. Endangered Species Act, a result of the ongoing loss of critical habitat by melting sea ice, polar bears have limited options for long term survival in the wild. Our current regime for safeguarding animals facing extinction frequently utilizes zoos and their breeding programs, to keep alive the genetic diversity of the species until some future time when reintroduction, somewhere, may be attempted. By exploring the potential for polar bears to be relocated to Antarctica, we must analyze the complex ethical, legal, political, financial, and biological realms, which are the backdrop to framing all questions in this arena. Can we do it? Should we do it? By utilizing an environmental ethics perspective, we propose that the Ecological Commons of the Arctic and Antarctic should not be viewed solely through the lens of human resource management needs. From this perspective, polar bears do not need our permission, they need our assistance. Antarctica therefore represents a second, if imperfect chance, to buy time for polar bears, in a world where polar regimes, not yet fully understood, are themselves quickly changing as a result of climate change.

Keywords: polar bear, climate change, environmental ethics, Arctic, Antarctica, assisted colonization, treaty

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515 Recognition of Sanitation as a Human Right: An Overview of Unresolutions and Reports That Recognizes the Human Right to Sanitation in South-Asian Countries

Authors: Anju Vaidya

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Sanitation is concerned with proper disposal of human excreta, waste water and promotion of hygiene. Lack of sanitation impacts our environment affecting our finance, schooling, health, and thus exacerbating poverty, discrimination and exclusion of the marginalized group. Sanitation can be a route and one of the most important factor to reach the goals of all Millennium Development goals. This study aims at exploring what are the rights to sanitation of the people, how it is enacted and what challenges are being faced while implementing the right to sanitation in South-Asian countries (India, Nepal, Pakistan, Bangladesh, Srilanka) at government, non-government and international level. This study also aims at finding how right sanitation is interlinked with children rights. The available reports submitted by government and civil society organizations working in South-Asian countries from the website of the Office of High Commissioner for Human Rights that were submitted under International covenant on economic, social and cultural rights and Convention on rights of the child have been selected and analyzed. The study uses Literature review to analyze these UN documents submitted from 2000 to 2015 in the context of South-Asian countries. Preliminary insight reveals that sanitation is recognized as one of the important factor to attain adequate standard of living. It has been found that inadequate sanitation has been a major factor that affects all aspects of life and one of its devastating impacts is increased child mortality. Many efforts have been made at national and international level in South-Asian countries to improve the state of sanitation and sanitation services. Various approaches such as Community led Total Sanitation, School led Total Sanitation, establishing Open Defecation free zone, water supply services and other sanitation and hygiene awareness programs are being launched. Despite different efforts and programs being implemented, sanitation and hygiene practices and behavior change remains to be a big challenge. Disparity in access and imbalance between urban and rural services and geographical regions, inadequate financing, clear policy framework and fragile functionality are some of the significant challenges faced while implementing these programs. Children are one of the most vulnerable group that are affected to a large extent. The study brings into light varied approaches that are being made and challenges that are being faced by government, non-government and civil society organizations while implementing the programs and strategies related to sanitation. It also highlights the relation of sanitation as a human right with child rights. This can help the stakeholders and policymakers better understand that improving sanitation situation is a process that requires learning, planning and behavior change and achieving sanitation coverage targets and motivating behavior change requires additional tools based on participation, non-discrimination and process approaches for planning and feedback.

Keywords: challenges, child rights, open defecation, sanitation as a human right

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514 Effect of Dietary Inclusion of Moringa oleifera Leaf Meal on Blood Biochemical Changes and Lipid Profile of Vanaraja Chicken in Tropics

Authors: Kaushalendra Kumar, Abhishek Kumar, Chandra Moni, Sanjay Kumar, P. K. Singh, Ajeet Kumar

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Present study investigated the dietary inclusion of Moringa oleifera leaf meal (MOLM) on production efficiency, hemato-biochemical profile and economy of Vanaraja birds under tropical condition. Experiment was conducted for a period of 56 days on 300 Vanaraja birds randomly divided in to five different experimental groups including control of 60 birds each group replicated with 20 chicks in each replicate. T1, T2, T3, T4, and T5 were offered with 0, 5, 10, 15, and 20% Moringa oleifera leaf meal along with basal ration. All the standard managemental practices were followed during experimental period including vaccination schedule. Locally available Moringa oleifera leaves were harvested at mature stage and allowed to dry under shady and aerated conditions. Thereafter, dried leaves were milled to make a leaf meal and stored in the airtight nylon bags to avoid any possible contamination from foreign material and use for experiment. Production parameters were calculated based on the amount of feed consumed and weight gain every weeks. The body weight gain of T2 group was significantly (P < 0.05) higher side whereas T3 group was comparable with control. The feed conversion ratio for T2 group was found to be significantly (P < 0.05) lower than all other treatment groups, while none of the group was comparable with each other. At the end of the experiment blood samples were collected from birds for haematology study while serum biochemistry performed using spectrophotometer following statndard protocols. The haematological attributes were significantly (P > 0.05) not differed among the groups. However, serum biochemistry showed significant reduction (P < 0.05) of blood urea nitrogen, uric acid and creatinine level with higher level of MOLM diet, indicates better utilization of protein supplemented through MOLM. The total cholesterol and triglyceride level was declined significantly (P < 0.05) as compare to control group with increased level of MOLM in basal diet, decreasing trend of serum cholesterol noted. However, value of HDL for T3 group was highest and for T1 group was lowest but no significant difference (P < 0.05) found among the groups. It might be due to presence of β-sitosterol a bioactive compound present in MOLM which causes lowering of plasma concentration of LDL. During experiment total, LDL and VLDL level was found to be decreased significantly (P < 0.05) as compare to control group. It was observed that the production efficiency of birds significantly improved with 5% followed by 10% Moringa oleifera leaf meal among the treatment groups. However, the maximum profit per kg live weight was noted in 10 % level and least profit observed in 20% MOLM fed group. It was concluded that the dietary inclusion of MOLM improved overall performances without affecting metabolic status and effective in reducing cholesterol level reflects healthy chicken production for human consumption.

Keywords: hemato biochemistry, Moringa oleifera leaf meal, performance, Vanaraja birds

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513 Impact of Electric Field on the Optical Properties of Hydrophilic Quantum Dots

Authors: Valentina V. Goftman, Vladislav A. Pankratov, Alexey V. Markin, Tangi Aubert, Zeger Hens, Sarah De Saeger, Irina Yu. Goryacheva

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The most important requirements for biochemical applicability of quantum dots (QDs) are: 1) the surface cap should render intact or improved optical properties; 2) mono-dispersion and good stability in aqueous phase in a wide range of pH and ionic strength values; 3) presence of functional groups, available for bioconjugation; 4) minimal impact from the environment on the QDs’ properties and, vice versa, minimal influence of the QDs’ components on the environment; and 5) stability against chemical/biochemical/physical influence. The latter is especially important for in vitro and in vivo applications. For example, some physical intracellular delivery strategies (e.g., electroporation) imply a rapid high-voltage electric field impulse in order to temporarily generate hydrophilic pores in the cell plasma membrane, necessary for the passive transportation of QDs into the cell. In this regard, it is interesting to investigate how different capping layers, which can provide high stability and sufficient fluorescent properties of QDs in a water solution, behave under these abnormal conditions. In this contribution, hydrophobic core-shell CdSe/CdS/CdZnS/ZnS QDs (λem=600 nm), produced by means of the Successive Ion Layer Adsorption and Reaction (SILAR) technique, were transferred to a water solution using two of the most commonly used methods: (i) encapsulation in an amphiphilic brush polymer based on poly(maleic anhydride-alt-1-octadecene) (PMAO) modified with polyethylene glycol (PEG) chains and (ii) silica covering. Polymer encapsulation preserves the initial ligands on the QDs’ surface owing to the hydrophobic attraction between the hydrophobic groups of the amphiphilic molecules and the surface hydrophobic groups of the QDs. This covering process allows maintaining the initial fluorescent properties, but it leads to a considerable increase of the QDs’ size. However, covering with a silica shell, by means of the reverse microemulsion method, allows maintaining both size and fluorescent properties of the initial QDs. The obtained water solutions of polymer covered and silica-coated QDs in three different concentrations were exposed to a low-voltage electric field for a short time and the fluorescent properties were investigated. It is shown that the PMAO-PEG polymer acquires some additional charges in the presence of the electric field, which causes repulsion between the polymer and the QDs’ surface. This process destroys the homogeneity of the whole amphiphilic shell and it dramatically decreases the fluorescent properties (dropping to 10% from its initial value) because of the direct contact of the QDs with the strongly oxidative environment (water). In contrast, a silica shell possesses dielectric properties which allow retaining 90% of its initial fluorescence intensity, even after a longer electric impact. Thus, silica shells are clearly a preferable covering for bio-application of QDs, because – besides the high uniform morphology, controlled size and biocompatibility – it allows protecting QDs from oxidation, even under the influence of an electric field.

Keywords: electric field, polymer coating, quantum dots, silica covering, stability

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512 Globalisation and Diplomacy: How Can Small States Improve the Practice of Diplomacy to Secure Their Foreign Policy Objectives?

Authors: H. M. Ross-McAlpine

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Much of what is written on diplomacy, globalization and the global economy addresses the changing nature of relationships between major powers. While the most dramatic and influential changes have resulted from these developing relationships the world is not, on deeper inspection, governed neatly by major powers. Due to advances in technology, the shifting balance of power and a changing geopolitical order, small states have the ability to exercise a greater influence than ever before. Increasingly interdependent and ever complex, our world is too delicate to be handled by a mighty few. The pressure of global change requires small states to adapt their diplomatic practices and diversify their strategic alliances and relationships. The nature and practice of diplomacy must be re-evaluated in light of the pressures resulting from globalization. This research examines: how small states can best secure their foreign policy objectives? Small state theory is used as a foundation for exploring the case study of New Zealand. The research draws on secondary sources to evaluate the existing theory in relation to modern practices of diplomacy. As New Zealand lacks the required economic and military power to play an active, influential role in international affairs what strategies are used to exert influence? Furthermore, New Zealand lies in a remote corner of the Pacific and is geographically isolated from its nearest neighbors how does this affect security and trade priorities? The findings note a significant shift since the 1970’s in New Zealand’s diplomatic relations. This shift is arguably a direct result of globalization, regionalism and a growing independence from the traditional bi-lateral relationships. The need to source predictable trade, investment and technology are an essential driving force for New Zealand’s diplomatic relations. A lack of hard power aligns New Zealand’s prosperity with a secure, rules-based international system that increases the likelihood of a stable and secure global order. New Zealand’s diplomacy and prosperity has been intrinsically reliant on its reputation. A vital component of New Zealand’s diplomacy is preserving a reputation for integrity and global responsibility. It is the use of this soft power that facilitates the influence that New Zealand enjoys on the world stage. To weave a comprehensive network of successful diplomatic relationships, New Zealand must maintain a reputation of international credibility. Globalization has substantially influenced the practice of diplomacy for New Zealand. The current world order places economic and military might in the hands of a few, subsequently requiring smaller states to use other means for securing their interests. There are clear strategies evident in New Zealand’s diplomacy practice that draw attention to how other smaller states might best secure their foreign policy objectives. While these findings are limited, as with all case study research, there is value in applying the findings to other small states struggling to secure their interests in the wake of rapid globalization.

Keywords: diplomacy, foreign policy, globalisation, small state

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511 Sustainability and Smart Cities Planning in Contrast with City Humanity. Human Scale and City Soul (Neighbourhood Scale)

Authors: Ghadir Hummeid

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Undoubtedly, our world is leading all the purposes and efforts to achieve sustainable development in life in all respects. Sustainability has been regarded as a solution to many challenges of our world today, materiality and immateriality. With the new consequences and challenges our world today, such as global climate change, the use of non-renewable resources, environmental pollution, the decreasing of urban health, the urban areas’ aging, the highly increasing migrations into urban areas linked to many consequences such as highly infrastructure density, social segregation. All of that required new forms of governance, new urban policies, and more efficient efforts and urban applications. Based on the fact that cities are the core of life and it is a fundamental life axis, their development can increase or decrease the life quality of their inhabitants. Architects and planners see themselves today in the need to create new approaches and new sustainable policies to develop urban areas to correspond with the physical and non-physical transformations that cities are nowadays experiencing. To enhance people's lives and provide for their needs in this present without compromising the needs and lives of future generations. The application of sustainability has become an inescapable part of the development and projections of cities' planning. Yet its definition has been indefinable due to the plurality and difference of its applications. As the conceptualizations of technology are arising and have dominated all life aspects today, from smart citizens and smart life rhythms to smart production and smart structures to smart frameworks, it has influenced the sustainability applications as well in the planning and urbanization of cities. The term "smart city" emerged from this influence as one of the possible key solutions to sustainability. The term “smart city” has various perspectives of applications and definitions in the literature and in urban applications. However, after the observation of smart city applications in current cities, this paper defined the smart city as an urban environment that is controlled by technologies yet lacks the physical architectural representation of this smartness as the current smart applications are mostly obscured from the public as they are applied now on a diminutive scale and highly integrated into the built environment. Regardless of the importance of these technologies in improving the quality of people's lives and in facing cities' challenges, it is important not to neglect their architectural and urban presentations will affect the shaping and development of city neighborhoods. By investigating the concept of smart cities and exploring its potential applications on a neighbourhood scale, this paper aims to shed light on understanding the challenges faced by cities and exploring innovative solutions such as smart city applications in urban mobility and how they affect the different aspects of communities. The paper aims to shape better articulations of smart neighborhoods’ morphologies on the social, architectural, functional, and material levels. To understand how to create more sustainable and liveable future approaches to developing urban environments inside cities. The findings of this paper will contribute to ongoing discussions and efforts in achieving sustainable urban development.

Keywords: sustainability, urban development, smart city, resilience, sense of belonging

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510 Development of a Miniature Laboratory Lactic Goat Cheese Model to Study the Expression of Spoilage by Pseudomonas Spp. In Cheeses

Authors: Abirami Baleswaran, Christel Couderc, Loubnah Belahcen, Jean Dayde, Hélène Tormo, Gwénaëlle Jard

Abstract:

Cheeses are often reported to be spoiled by Pseudomonas spp., responsible for defects in appearance, texture, taste, and smell, leading to their non-marketing and even their destruction. Despite preventive actions, problems linked to Pseudomonas spp. are difficult to control by the lack of knowledge and control of these contaminants during the cheese manufacturing. Lactic goat cheese producers are not spared by this problem and are looking for solutions to decrease the number of spoiled cheeses. To explore different hypotheses, experiments are needed. However, cheese-making experiments at the pilot scale are expensive and time consuming. Thus, there is a real need to develop a miniature cheeses model system under controlled conditions. In a previous study, several miniature cheese models corresponding to different type of commercial cheeses have been developed for different purposes. The models were, for example, used to study the influence of milk, starters cultures, pathogen inhibiting additives, enzymatic reactions, microflora, freezing process on cheese. Nevertheless, no miniature model was described on the lactic goat cheese. The aim of this work was to develop a miniature cheese model system under controlled laboratory conditions which resembles commercial lactic goat cheese to study Pseudomonas spp. spoilage during the manufacturing and ripening process. First, a protocol for the preparation of miniature cheeses (3.5 times smaller than a commercial one) was designed based on the cheese factorymanufacturing process. The process was adapted from “Rocamadour” technology and involves maturation of pasteurized milk, coagulation, removal of whey by centrifugation, moulding, and ripening in a little scale cellar. Microbiological (total bacterial count, yeast, molds) and physicochemical (pH, saltinmoisture, moisture in fat-free)analyses were performed on four key stages of the process (before salting, after salting, 1st day of ripening, and end of ripening). Factory and miniature cheeses volatilomewere also obtained after full scan Sift-MS cheese analysis. Then, Pseudomonas spp. strains isolated from contaminated cheeses were selected on their origin, their ability to produce pigments, and their enzymatic activities (proteolytic, lecithinasic, and lipolytic). Factory and miniature curds were inoculated by spotting selected strains on the cheese surface. The expression of cheese spoilage was evaluated by counting the level of Pseudomonas spp. during the ripening and by visual observation and under UVlamp. The physicochemical and microbiological compositions of miniature cheeses permitted to assess that miniature process resembles factory process. As expected, differences involatilomes were observed, probably due to the fact that miniature cheeses are made usingpasteurized milk to better control the microbiological conditions and also because the little format of cheese induced probably a difference during the ripening even if the humidity and temperature in the cellar were quite similar. The spoilage expression of Pseudomonas spp. was observed in miniature and factory cheeses. It confirms that the proposed model is suitable for the preparation of miniature cheese specimens in the spoilage study of Pseudomonas spp. in lactic cheeses. This kind of model could be deployed for other applications and other type of cheese.

Keywords: cheese, miniature, model, pseudomonas spp, spoilage

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509 Use of Proton Pump Inhibitors Medications during the First Years of Life and Late Complications

Authors: Kamelia Hamza

Abstract:

Background: Proton pump inhibitors (PPIs) are the most prescribed drug classes for pediatric gastroesophageal reflux disease (GERD).Many patients are treated with these drugs for atypical manifestations attributed to gastroesophageal reflux (GER), even in the absence of proved causal relationship. There is an impression of increase use of PPI's treatment for reflux in "clalit health services," the largest health organization in Israel. In the recent years, the medicine is given without restriction, it's not limited to pediatric gastroenterologists only, but pediatricians and family doctors. The objective of this study is to evaluate the hypothesis that exposure to PPIs during the first year of life is associated with an increased risk of developing late adverse diseases: pneumonia, asthma, AGE, IBD, celiac disease, allergic disorders, obesity, attention deficit hyperactivity disorders (ADHD), autism spectrum disorders (ASD). Methods: The study is a retrospective case-control cohort study based on a computerized database of Clalit Health Services (CHS). It includes 9844 children born between 2002-2018 and reported to complain of at least one of the symptoms (reflux/ spitting up, irritability, feeding difficulties, colics). The study population included the study group (n=4922) of children exposed to PPIs at any time prior to the first year of life and a control group (n=4922) child not exposed to PPIs who were matched to each case of the study group on age, race, socioeconomic status, and year of birth. The prevalence of late complications/diseases in the study group was compared with the prevalence of late complications/diseases diagnosis between 2002-2020 in the control group. Odds ratios and 95% confidence intervals were calculated by using logistic regression models. Results: We found that compared to the control group, children exposed to PPIs in the first year of life had an increased risk of developing several late complications/ disorders: pneumonia, asthma, various allergies (urticaria, allergic rhinitis, or allergic conjunctivitis) OR, inhalant allergies, and food allergies. In addition, they showed an increased risk of being diagnosed with ADHD or ASD, but children exposed to PPIs in the first year of life had decrease the risk of obesity by 17% (OR 0.825, 95%CI 0.697-0.976). Conclusions: We found significant associations between the use of PPIs during the first year of life and subsequent development of late complications/diseases such as respiratory diseases, allergy diseases, ADHD, and ASD. More studies are needed to prove causality and determine the mechanism behind the effect of PPIs and the development of late complications.

Keywords: acid suppressing medications, proton pump inhibitors, histamine 2 blocker, late complications, gastroesophageal reflux, gastroesophageal reflux disease, acute gastroenteritis, community acquired pneumonia, asthma, allergic diseases, obesity, inflammatory bowel diseases, ulcerative colitis, crohn disease, attention deficit hyperactivity disorders, autism spectrum disorders

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508 Electricity Market Reforms Towards Clean Energy Transition andnd Their Impact in India

Authors: Tarun Kumar Dalakoti, Debajyoti Majumder, Aditya Prasad Das, Samir Chandra Saxena

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India’s ambitious target to achieve a 50 percent share of energy from non-fossil fuels and the 500-gigawatt (GW) renewable energy capacity before the deadline of 2030, coupled with the global pursuit of sustainable development, will compel the nation to embark on a rapid clean energy transition. As a result, electricity market reforms will emerge as critical policy instruments to facilitate this transition and achieve ambitious environmental targets. This paper will present a comprehensive analysis of the various electricity market reforms to be introduced in the Indian Electricity sector to facilitate the integration of clean energy sources and will assess their impact on the overall energy landscape. The first section of this paper will delve into the policy mechanisms to be introduced by the Government of India and the Central Electricity Regulatory Commission to promote clean energy deployment. These mechanisms include extensive provisions for the integration of renewables in the Indian Electricity Grid Code, 2023. The section will also cover the projection of RE Generation as highlighted in the National Electricity Plan, 2023. It will discuss the introduction of Green Energy Market segments, the waiver of Inter-State Transmission System (ISTS) charges for inter-state sale of solar and wind power, the notification of Promoting Renewable Energy through Green Energy Open Access Rules, and the bundling of conventional generating stations with renewable energy sources. The second section will evaluate the tangible impact of these electricity market reforms. By drawing on empirical studies and real-world case examples, the paper will assess the penetration rate of renewable energy sources in India’s electricity markets, the decline of conventional fuel-based generation, and the consequent reduction in carbon emissions. Furthermore, it will explore the influence of these reforms on electricity prices, the impact on various market segments due to the introduction of green contracts, and grid stability. The paper will also discuss the operational challenges to be faced due to the surge of RE Generation sources as a result of the implementation of the above-mentioned electricity market reforms, including grid integration issues, intermittency concerns with renewable energy sources, and the need for increasing grid resilience for future high RE in generation mix scenarios. In conclusion, this paper will emphasize that electricity market reforms will be pivotal in accelerating the global transition towards clean energy systems. It will underscore the importance of a holistic approach that combines effective policy design, robust regulatory frameworks, and active participation from market actors. Through a comprehensive examination of the impact of these reforms, the paper will shed light on the significance of India’s sustained commitment to a cleaner, more sustainable energy future.

Keywords: renewables, Indian electricity grid code, national electricity plan, green energy market

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507 Development of Alternative Fuels Technologies for Transportation

Authors: Szymon Kuczynski, Krystian Liszka, Mariusz Laciak, Andrii Oliinyk, Adam Szurlej

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Currently, in automotive transport to power vehicles, almost exclusively hydrocarbon based fuels are used. Due to increase of hydrocarbon fuels consumption, quality parameters are tightend for clean environment. At the same time efforts are undertaken for development of alternative fuels. The reasons why looking for alternative fuels for petroleum and diesel are: to increase vehicle efficiency and to reduce the environmental impact, reduction of greenhouse gases emissions and savings in consumption of limited oil resources. Significant progress was performed on development of alternative fuels such as methanol, ethanol, natural gas (CNG / LNG), LPG, dimethyl ether (DME) and biodiesel. In addition, biggest vehicle manufacturers work on fuel cell vehicles and its introduction to the market. Alcohols such as methanol and ethanol create the perfect fuel for spark-ignition engines. Their advantages are high-value antiknock which determines their application as additive (10%) to unleaded petrol and relative purity of produced exhaust gasses. Ethanol is produced in distillation process of plant products, which value as a food can be irrational. Ethanol production can be costly also for the entire economy of the country, because it requires a large complex distillation plants, large amounts of biomass and finally a significant amount of fuel to sustain the process. At the same time, the fermentation process of plants releases into the atmosphere large quantities of carbon dioxide. Natural gas cannot be directly converted into liquid fuels, although such arrangements have been proposed in the literature. Going through stage of intermediates is inevitable yet. Most popular one is conversion to methanol, which can be processed further to dimethyl ether (DME) or olefin (ethylene and propylene) for the petrochemical sector. Methanol uses natural gas as a raw material, however, requires expensive and advanced production processes. In relation to pollution emissions, the optimal vehicle fuel is LPG which is used in many countries as an engine fuel. Production of LPG is inextricably linked with production and processing of oil and gas, and which represents a small percentage. Its potential as an alternative for traditional fuels is therefore proportionately reduced. Excellent engine fuel may be biogas, however, follows to the same limitations as ethanol - the same production process is used and raw materials. Most essential fuel in the campaign of environment protection against pollution is natural gas. Natural gas as fuel may be either compressed (CNG) or liquefied (LNG). Natural gas can also be used for hydrogen production in steam reforming. Hydrogen can be used as a basic starting material for the chemical industry, an important raw material in the refinery processes, as well as a fuel vehicle transportation. Natural gas can be used as CNG which represents an excellent compromise between the availability of the technology that is proven and relatively cheap to use in many areas of the automotive industry. Natural gas can also be seen as an important bridge to other alternative sources of energy derived from fuel and harmless to the environment. For these reasons CNG as a fuel stimulates considerable interest in the worldwide.

Keywords: alternative fuels, CNG (Compressed Natural Gas), LNG (Liquefied Natural Gas), NGVs (Natural Gas Vehicles)

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506 The Study of Fine and Nanoscale Gold in the Ores of Primary Deposits and Gold-Bearing Placers of Kazakhstan

Authors: G. Omarova, S. Assubayeva, S. Tugambay, K. Bulegenov

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The article discusses the problem of developing a methodology for studying thin and nanoscale gold in ores and placers of primary deposits, which will allow us to develop schemes for revealing dispersed gold inclusions and thus improve its recovery rate to increase the gold reserves of the Republic of Kazakhstan. The type of studied gold, is characterized by a number of features. In connection with this, the conditions of its concentration and distribution in ore bodies and formations, as well as the possibility of reliably determining it by "traditional" methods, differ significantly from that of fine gold (less than 0.25 microns) and even more so from that of larger grains. The mineral composition of rocks (metasomatites) and gold ore and the mineralization associated with them was studied in detail on the Kalba ore field in Kazakhstan. Mineralized zones were identified and samples were taken from them for analytical studies. The research revealed paragenetic relationships of newly formed mineral formations at the nanoscale, which makes it possible to clarify the conditions for formation of deposits with a particular type of mineralization. This will provide significant assistance in developing a scheme for study. Typomorphic features of gold were revealed, and mechanisms of formation and aggregation of gold nanoparticles were proposed. The presence of a large number of particles isolated at the laboratory stage from concentrates of gravitational enrichment can serve as an indicator of the presence of even smaller particles in the object. Even the most advanced devices based on gravitational methods for gold concentration provide extraction of metal at a level of around 50%, while pulverized metal is extracted much worse, and gold of less than 1 micron size is extracted at only a few percent. Therefore, when particles of gold smaller than 10 microns are detected, their actual numbers may be significantly higher than expected. In particular, at the studied sites, enrichment of slurry and samples with volumes up to 1 m³ was carried out using a screw lock or separator to produce a final concentrate weighing up to several kilograms. Free gold particles were extracted from the concentrates in the laboratory using a number of processes (magnetic and electromagnetic separation, washing with bromoform in a cup to obtain an ultracontentrate, etc.) and examined under electron microscopes to investigate the nature of their surface and chemical composition. The main result of the study was the detection of gold nanoparticles located on the surface of loose metal grains. The most characteristic forms of gold secretions are individual nanoparticles and aggregates of different configurations. Sometimes, aggregates form solid dense films, deposits, and crusts, all of which are confined to the negative forms of the nano- and microrelief on the surfaces of golden. The results will provide significant knowledge about the prevalence and conditions for the distribution of fine and nanoscale gold in Kazakhstan deposits, as well as the development of methods for studying it, which will minimize losses of this type of gold during extraction.

Keywords: electron microscopy, microminerology, placers, thin and nanoscale gold

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505 Safety of Mesenchymal Stem Cells Therapy: Potential Risk of Spontaneous Transformations

Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska

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Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.

Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture

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504 Gold Nanoprobes Assay for the Identification of Foodborn Pathogens Such as Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

Abstract:

Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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