Search results for: reactive molecular dynamics
Commenced in January 2007
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Edition: International
Paper Count: 5335

Search results for: reactive molecular dynamics

265 2,7-Diazaindole as a Photophysical Probe for Excited State Hydrogen/Proton Transfer

Authors: Simran Baweja, Bhavika Kalal, Surajit Maity

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Photoinduced tautomerization reactions have been the centre of attention among the scientific community over the past several decades because of their significance in various biological systems. 7-azaindole (7AI) is considered a model system for DNA base pairing and to understand the role of such tautomerization reactions in mutations. To the best of our knowledge, extensive studies have been carried out on 7-azaindole and its solvent clusters exhibiting proton/ hydrogen transfer in both solution as well as gas phases. Derivatives of the above molecule, like 2,7- and 2,6-diazaindoles are proposed to have even better photophysical properties due to the presence of -aza group on the 2nd position. However, there are studies in the solution phase that suggest the relevance of these molecules, but there are no experimental studies reported in the gas phase yet. In our current investigation, we present the first gas phase spectroscopic data of 2,7-diazaindole (2,7-DAI) and its solvent cluster (2,7-DAI-H2O). In this, we have employed state-of-the-art laser spectroscopic methods such as fluorescence excitation (LIF), dispersed fluorescence (DF), resonant two-photon ionization-time of flight mass spectrometry (2C-R2PI), photoionization efficiency spectroscopy (PIE), IR-UV double resonance spectroscopy, i.e., fluorescence-dip infrared spectroscopy (FDIR) and resonant ion-dip infrared spectroscopy (IDIR) to understand the electronic structure of the molecule. The origin band corresponding to the S1 ← S0 transition of the bare 2,7-DAI is found to be positioned at 33910 cm-1, whereas the origin band corresponding to S1 ← S0 transition of the 2,7-DAI-H2O is positioned at 33074 cm-1. The red-shifted transition in the case of solvent cluster suggests the enhanced feasibility of excited state hydrogen/ proton transfer. The ionization potential for the 2,7-DAI molecule is found to be 8.92 eV which is significantly higher than the previously reported 7AI (8.11 eV) molecule, making it a comparatively complex molecule to study. The ionization potential is reduced by 0.14 eV in the case of 2,7-DAI-H2O (8.78 eV) cluster compared to that of 2,7-DAI. Moreover, on comparison with the available literature values of 7AI, we found the origin band of 2,7-DAI and 2,7-DAI-H2O to be red-shifted by -729 and -280 cm-1 respectively. The ground and excited state N-H stretching frequencies of the 27DAI molecule were determined using fluorescence-dip infrared spectra (FDIR) and resonant ion dip infrared spectroscopy (IDIR), obtained at 3523 and 3467 cm-1, respectively. The lower value of vNH in the electronically excited state of 27DAI implies the higher acidity of the group compared to the ground state. Moreover, we have done extensive computational analysis, which suggests that the energy barrier in the excited state reduces significantly as we increase the number of catalytic solvent molecules (S= H2O, NH3) as well as the polarity of solvent molecules. We found that the ammonia molecule is a better candidate for hydrogen transfer compared to water because of its higher gas-phase basicity. Further studies are underway to understand the excited state dynamics and photochemistry of such N-rich chromophores.

Keywords: excited state hydrogen transfer, supersonic expansion, gas phase spectroscopy, IR-UV double resonance spectroscopy, laser induced fluorescence, photoionization efficiency spectroscopy

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264 Detection and Identification of Antibiotic Resistant UPEC Using FTIR-Microscopy and Advanced Multivariate Analysis

Authors: Uraib Sharaha, Ahmad Salman, Eladio Rodriguez-Diaz, Elad Shufan, Klaris Riesenberg, Irving J. Bigio, Mahmoud Huleihel

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Antimicrobial drugs have played an indispensable role in controlling illness and death associated with infectious diseases in animals and humans. However, the increasing resistance of bacteria to a broad spectrum of commonly used antibiotics has become a global healthcare problem. Many antibiotics had lost their effectiveness since the beginning of the antibiotic era because many bacteria have adapted defenses against these antibiotics. Rapid determination of antimicrobial susceptibility of a clinical isolate is often crucial for the optimal antimicrobial therapy of infected patients and in many cases can save lives. The conventional methods for susceptibility testing require the isolation of the pathogen from a clinical specimen by culturing on the appropriate media (this culturing stage lasts 24 h-first culturing). Then, chosen colonies are grown on media containing antibiotic(s), using micro-diffusion discs (second culturing time is also 24 h) in order to determine its bacterial susceptibility. Other methods, genotyping methods, E-test and automated methods were also developed for testing antimicrobial susceptibility. Most of these methods are expensive and time-consuming. Fourier transform infrared (FTIR) microscopy is rapid, safe, effective and low cost method that was widely and successfully used in different studies for the identification of various biological samples including bacteria; nonetheless, its true potential in routine clinical diagnosis has not yet been established. The new modern infrared (IR) spectrometers with high spectral resolution enable measuring unprecedented biochemical information from cells at the molecular level. Moreover, the development of new bioinformatics analyses combined with IR spectroscopy becomes a powerful technique, which enables the detection of structural changes associated with resistivity. The main goal of this study is to evaluate the potential of the FTIR microscopy in tandem with machine learning algorithms for rapid and reliable identification of bacterial susceptibility to antibiotics in time span of few minutes. The UTI E.coli bacterial samples, which were identified at the species level by MALDI-TOF and examined for their susceptibility by the routine assay (micro-diffusion discs), are obtained from the bacteriology laboratories in Soroka University Medical Center (SUMC). These samples were examined by FTIR microscopy and analyzed by advanced statistical methods. Our results, based on 700 E.coli samples, were promising and showed that by using infrared spectroscopic technique together with multivariate analysis, it is possible to classify the tested bacteria into sensitive and resistant with success rate higher than 90% for eight different antibiotics. Based on these preliminary results, it is worthwhile to continue developing the FTIR microscopy technique as a rapid and reliable method for identification antibiotic susceptibility.

Keywords: antibiotics, E.coli, FTIR, multivariate analysis, susceptibility, UTI

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263 Fucoidan: A Potent Seaweed-Derived Polysaccharide with Immunomodulatory and Anti-inflammatory Properties

Authors: Tauseef Ahmad, Muhammad Ishaq, Mathew Eapen, Ahyoung Park, Sam Karpiniec, Vanni Caruso, Rajaraman Eri

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Fucoidans are complex, fucose-rich sulfated polymers discovered in brown seaweeds. Fucoidans are popular around the world, particularly in the nutraceutical and pharmaceutical industries, due to their promising medicinal properties. Fucoidans have been shown to have a variety of biological activities, including anti-inflammatory effects. They are known to inhibit inflammatory processes through a variety of mechanisms, including enzyme inhibition and selectin blockade. Inflammation is a part of the complicated biological response of living systems to damaging stimuli, and it plays a role in the pathogenesis of a variety of disorders, including arthritis, inflammatory bowel disease, cancer, and allergies. In the current investigation, various fucoidan extracts from Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica were assessed for inhibition of pro-inflammatory cytokine production (TNF-α, IL-1β, and IL-6) in LPS induced human macrophage cell line (THP-1) and human peripheral blood mononuclear cells (PBMCs). Furthermore, we also sought to catalogue these extracts based on their anti-inflammatory effects in the current in-vitro cell model. Materials and Methods: To assess the cytotoxicity of fucoidan extracts, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5, -diphenyltetrazolium bromide) cell viability assay was performed. Furthermore, a dose-response for fucoidan extracts was performed in LPS induced THP-1 cells and PBMCs after pre-treatment for 24 hours, and levels of TNF-α, IL-1β, and IL-6 cytokines were measured using Enzyme-Linked Immunosorbent Assay (ELISA). Results: The MTT cell viability assay demonstrated that fucoidan extracts exhibited no evidence of cytotoxicity in THP-1 cells or PBMCs after 48 hours of incubation. The results of the sandwich ELISA revealed that all fucoidan extracts suppressed cytokine production in LPS-stimulated PBMCs and human THP-1 cells in a dose-dependent manner. Notably, at lower concentrations, the lower molecular fucoidan (5-30 kDa) extract from Macrocystis pyrifera was a highly efficient inhibitor of pro-inflammatory cytokines. Fucoidan extracts from all species including Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica exhibited significant anti-inflammatory effects. These findings on several fucoidan extracts provide insight into strategies for improving their efficacy against inflammation-related diseases. Conclusion: In the current research, we have successfully catalogued several fucoidan extracts based on their efficiency in LPS-induced macrophages and PBMCs in downregulating the key pro-inflammatory cytokines (TNF-, IL-1 and IL-6), which are prospective targets in human inflammatory illnesses. Further research would provide more information on the mechanism of action, allowing it to be tested for therapeutic purposes as an anti-inflammatory medication.

Keywords: fucoidan, PBMCs, THP-1, TNF-α, IL-1β, IL-6, inflammation

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262 Reducing the Computational Cost of a Two-way Coupling CFD-FEA Model via a Multi-scale Approach for Fire Determination

Authors: Daniel Martin Fellows, Sean P. Walton, Jennifer Thompson, Oubay Hassan, Kevin Tinkham, Ella Quigley

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Structural integrity for cladding products is a key performance parameter, especially concerning fire performance. Cladding products such as PIR-based sandwich panels are tested rigorously, in line with industrial standards. Physical fire tests are necessary to ensure the customer's safety but can give little information about critical behaviours that can help develop new materials. Numerical modelling is a tool that can help investigate a fire's behaviour further by replicating the fire test. However, fire is an interdisciplinary problem as it is a chemical reaction that behaves fluidly and impacts structural integrity. An analysis using Computational Fluid Dynamics (CFD) and Finite Element Analysis (FEA) is needed to capture all aspects of a fire performance test. One method is a two-way coupling analysis that imports the updated changes in thermal data, due to the fire's behaviour, to the FEA solver in a series of iterations. In light of our recent work with Tata Steel U.K using a two-way coupling methodology to determine the fire performance, it has been shown that a program called FDS-2-Abaqus can make predictions of a BS 476 -22 furnace test with a degree of accuracy. The test demonstrated the fire performance of Tata Steel U.K Trisomet product, a Polyisocyanurate (PIR) based sandwich panel used for cladding. Previous works demonstrated the limitations of the current version of the program, the main limitation being the computational cost of modelling three Trisomet panels, totalling an area of 9 . The computational cost increases substantially, with the intention to scale up to an LPS 1181-1 test, which includes a total panel surface area of 200 .The FDS-2-Abaqus program is developed further within this paper to overcome this obstacle and better accommodate Tata Steel U.K PIR sandwich panels. The new developments aim to reduce the computational cost and error margin compared to experimental data. One avenue explored is a multi-scale approach in the form of Reduced Order Modeling (ROM). The approach allows the user to include refined details of the sandwich panels, such as the overlapping joints, without a computationally costly mesh size.Comparative studies will be made between the new implementations and the previous study completed using the original FDS-2-ABAQUS program. Validation of the study will come from physical experiments in line with governing body standards such as BS 476 -22 and LPS 1181-1. The physical experimental data includes the panels' gas and surface temperatures and mechanical deformation. Conclusions are drawn, noting the new implementations' impact factors and discussing the reasonability for scaling up further to a whole warehouse.

Keywords: fire testing, numerical coupling, sandwich panels, thermo fluids

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261 Developing a High Performance Cement Based Material: The Influence of Silica Fume and Organosilane

Authors: Andrea Cretu, Calin Cadar, Maria Miclaus, Lucian Barbu-Tudoran, Siegfried Stapf, Ioan Ardelean

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Additives and mineral admixtures have become an integral part of cement-based materials. It is common practice to add silica fume to cement based mixes in order to produce high-performance concrete. There is still a lack of scientific understanding regarding the effects that silica fume has on the microstructure of hydrated cement paste. The aim of the current study is to develop high-performance materials with low permeability and high resistance to flexural stress using silica fume and an organosilane. Organosilane bonds with cement grains and silica fume, influencing both the workability and the final properties of the mix, especially the pore size distributions and pore connectivity. Silica fume is a known pozzolanic agent which reacts with the calcium hydroxide in hydrated cement paste, producing more C-S-H and improving the mechanical properties of the mix. It is believed that particles of silica fume act as capillary pore fillers and nucleation centers for C-S-H and other hydration products. In order to be able to design cement-based materials with added silica fume and organosilane, it is necessary first to understand the formation of the porous network during hydration and to observe the distribution of pores and their connectivity. Nuclear magnetic resonance (NMR) methods in low-fields are non-destructive and allow the study of cement-based materials from the standpoint of their porous structure. Other methods, such as XRD and SEM-EDS, help create a comprehensive picture of the samples, along with the classic mechanical tests (compressive and flexural strength measurements). The transverse relaxation time (T₂) was measured during the hydration of 16 samples prepared with two water/cement ratios (0.3 and 0.4) and different concentrations or organosilane (APTES, up to 2% by mass of cement) and silica fume (up to 6%). After their hydration, the pore size distribution was assessed using the same NMR approach on the samples filled with cyclohexane. The SEM-EDS and XRD measurements were applied on pieces and powders prepared from the samples that were used in mechanical testing, which were kept under water for 28 days. Adding silica fume does not influence the hydration dynamics of cement paste, while the addition of organosilane extends the dormancy stage up to 10 hours. The size distribution of the capillary pores is not influenced by the addition of silica fume or organosilane, while the connectivity of capillary pores is decreased only when there is organosilane in the mix. No filling effect is observed even at the highest concentration of silica fume. There is an apparent increase in flexural strength of samples prepared only with silica fume and a decrease for those prepared with organosilane, with a few exceptions. XRD reveals that the pozzolanic reactivity of silica fume can only be observed when there is no organosilane present and the SEM-EDS method reveals the pore distribution, as well as hydration products and the presence or absence of calcium hydroxide. The current work was funded by the Romanian National Authority for Scientific Research, CNCS – UEFISCDI, through project PN-III-P2-2.1-PED-2016-0719.

Keywords: cement hydration, concrete admixtures, NMR, organosilane, porosity, silica fume

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260 Telomerase, a Biomarker in Oral Cancer Cell Proliferation and Tool for Its Prevention at Initial Stage

Authors: Shaista Suhail

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As cancer populations is increasing sharply, the incidence of oral squamous cell carcinoma (OSCC) has also been expected to increase. Oral carcinogenesis is a highly complex, multistep process which involves accumulation of genetic alterations that lead to the induction of proteins promoting cell growth (encoded by oncogenes), increased enzymatic (telomerase) activity promoting cancer cell proliferation. The global increase in frequency and mortality, as well as the poor prognosis of oral squamous cell carcinoma, has intensified current research efforts in the field of prevention and early detection of this disease. The advances in the understanding of the molecular basis of oral cancer should help in the identification of new markers. The study of the carcinogenic process of the oral cancer, including continued analysis of new genetic alterations, along with their temporal sequencing during initiation, promotion and progression, will allow us to identify new diagnostic and prognostic factors, which will provide a promising basis for the application of more rational and efficient treatments. Telomerase activity has been readily found in most cancer biopsies, in premalignant lesions or germ cells. Activity of telomerase is generally absent in normal tissues. It is known to be induced upon immortalization or malignant transformation of human cells such as in oral cancer cells. Maintenance of telomeres plays an essential role during transformation of precancer to malignant stage. Mammalian telomeres, a specialized nucleoprotein structures are composed of large conctamers of the guanine-rich sequence 5_-TTAGGG-3_. The roles of telomeres in regulating both stability of genome and replicative immortality seem to contribute in essential ways in cancer initiation and progression. It is concluded that activity of telomerase can be used as a biomarker for diagnosis of malignant oral cancer and a target for inactivation in chemotherapy or gene therapy. Its expression will also prove to be an important diagnostic tool as well as a novel target for cancer therapy. The activation of telomerase may be an important step in tumorgenesis which can be controlled by inactivating its activity during chemotherapy. The expression and activity of telomerase are indispensable for cancer development. There are no drugs which can effect extremely to treat oral cancers. There is a general call for new emerging drugs or methods that are highly effective towards cancer treatment, possess low toxicity, and have a minor environment impact. Some novel natural products also offer opportunities for innovation in drug discovery. Natural compounds isolated from medicinal plants, as rich sources of novel anticancer drugs, have been of increasing interest with some enzyme (telomerase) blockage property. The alarming reports of cancer cases increase the awareness amongst the clinicians and researchers pertaining to investigate newer drug with low toxicity.

Keywords: oral carcinoma, telomere, telomerase, blockage

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259 Performance Validation of Model Predictive Control for Electrical Power Converters of a Grid Integrated Oscillating Water Column

Authors: G. Rajapakse, S. Jayasinghe, A. Fleming

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This paper aims to experimentally validate the control strategy used for electrical power converters in grid integrated oscillating water column (OWC) wave energy converter (WEC). The particular OWC’s unidirectional air turbine-generator output power results in discrete large power pulses. Therefore, the system requires power conditioning prior to integrating to the grid. This is achieved by using a back to back power converter with an energy storage system. A Li-Ion battery energy storage is connected to the dc-link of the back-to-back converter using a bidirectional dc-dc converter. This arrangement decouples the system dynamics and mitigates the mismatch between supply and demand powers. All three electrical power converters used in the arrangement are controlled using finite control set-model predictive control (FCS-MPC) strategy. The rectifier controller is to regulate the speed of the turbine at a set rotational speed to uphold the air turbine at a desirable speed range under varying wave conditions. The inverter controller is to maintain the output power to the grid adhering to grid codes. The dc-dc bidirectional converter controller is to set the dc-link voltage at its reference value. The software modeling of the OWC system and FCS-MPC is carried out in the MATLAB/Simulink software using actual data and parameters obtained from a prototype unidirectional air-turbine OWC developed at Australian Maritime College (AMC). The hardware development and experimental validations are being carried out at AMC Electronic laboratory. The designed FCS-MPC for the power converters are separately coded in Code Composer Studio V8 and downloaded into separate Texas Instrument’s TIVA C Series EK-TM4C123GXL Launchpad Evaluation Boards with TM4C123GH6PMI microcontrollers (real-time control processors). Each microcontroller is used to drive 2kW 3-phase STEVAL-IHM028V2 evaluation board with an intelligent power module (STGIPS20C60). The power module consists of a 3-phase inverter bridge with 600V insulated gate bipolar transistors. Delta standard (ASDA-B2 series) servo drive/motor coupled to a 2kW permanent magnet synchronous generator is served as the turbine-generator. This lab-scale setup is used to obtain experimental results. The validation of the FCS-MPC is done by comparing these experimental results to the results obtained by MATLAB/Simulink software results in similar scenarios. The results show that under the proposed control scheme, the regulated variables follow their references accurately. This research confirms that FCS-MPC fits well into the power converter control of the OWC-WEC system with a Li-Ion battery energy storage.

Keywords: dc-dc bidirectional converter, finite control set-model predictive control, Li-ion battery energy storage, oscillating water column, wave energy converter

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258 An Adaptable Semi-Numerical Anisotropic Hyperelastic Model for the Simulation of High Pressure Forming

Authors: Daniel Tscharnuter, Eliza Truszkiewicz, Gerald Pinter

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High-quality surfaces of plastic parts can be achieved in a very cost-effective manner using in-mold processes, where e.g. scratch resistant or high gloss polymer films are pre-formed and subsequently receive their support structure by injection molding. The pre-forming may be done by high-pressure forming. In this process, a polymer sheet is heated and subsequently formed into the mold by pressurized air. Due to the heat transfer to the cooled mold the polymer temperature drops below its glass transition temperature. This ensures that the deformed microstructure is retained after depressurizing, giving the sheet its final formed shape. The development of a forming process relies heavily on the experience of engineers and trial-and-error procedures. Repeated mold design and testing cycles are however both time- and cost-intensive. It is, therefore, desirable to study the process using reliable computer simulations. Through simulations, the construction of the mold and the effect of various process parameters, e.g. temperature levels, non-uniform heating or timing and magnitude of pressure, on the deformation of the polymer sheet can be analyzed. Detailed knowledge of the deformation is particularly important in the forming of polymer films with integrated electro-optical functions. Care must be taken in the placement of devices, sensors and electrical and optical paths, which are far more sensitive to deformation than the polymers. Reliable numerical prediction of the deformation of the polymer sheets requires sophisticated material models. Polymer films are often either transversely isotropic or orthotropic due to molecular orientations induced during manufacturing. The anisotropic behavior affects the resulting strain field in the deformed film. For example, parts of the same shape but different strain fields may be created by varying the orientation of the film with respect to the mold. The numerical simulation of the high-pressure forming of such films thus requires material models that can capture the nonlinear anisotropic mechanical behavior. There are numerous commercial polymer grades for the engineers to choose from when developing a new part. The effort required for comprehensive material characterization may be prohibitive, especially when several materials are candidates for a specific application. We, therefore, propose a class of models for compressible hyperelasticity, which may be determined from basic experimental data and which can capture key features of the mechanical response. Invariant-based hyperelastic models with a reduced number of invariants are formulated in a semi-numerical way, such that the models are determined from a single uniaxial tensile tests for isotropic materials, or two tensile tests in the principal directions for transversely isotropic or orthotropic materials. The simulation of the high pressure forming of an orthotropic polymer film is finally done using an orthotropic formulation of the hyperelastic model.

Keywords: hyperelastic, anisotropic, polymer film, thermoforming

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257 Journeys of Healing for Military Veterans: A Pilot Study

Authors: Heather Warfield, Brad Genereux

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Military personnel encounter a number of challenges when separating from military service to include career uncertainty, relational/family dynamics, trauma as a result of military experiences, reconceptualization of identity, and existential issues related to purpose, meaning making and framing of the military experience(s). Embedded within military culture are well-defined rites of passage and a significant sense of belonging. Consequently, transition out of the military can result in the loss of such rites of passage and belongingness. However, a pilgrimage journey can provide the time and space to engage in a new rite of passage, to construct a new pilgrim identity, and a to develop deep social relationships that lead to a sense of belongingness to a particular pilgrim community as well as to the global community of pilgrims across numerous types of pilgrimage journeys. The aims of the current paper are to demonstrate the rationale for why pilgrimage journeys are particularly significant for military veterans, provide an overview of an innovative program that facilitates the Camino de Santiago pilgrimage for military veterans, and discusses the lessons learned from the initial pilot project of a recently established program. Veterans on the Camino (VOC) is an emerging nongovernmental organization in the USA. Founded by a military veteran, after leaving his military career, the primary objective of the organization is to facilitate healing for veterans via the Camino de Santiago pilgrimage journey. As part of the program, participants complete a semi-structured interview at three time points – pre, during, and post journey. The interview items are based on ongoing research by the principal investigator and address such constructs as meaning-making, wellbeing, therapeutic benefits and transformation. In addition, program participants complete The Sources of Meaning and Meaning in Life Questionnaire (SoMe). The pilot program occurred in the spring of 2017. Five participants were selected after an extensive application process and review by a three-person selection board. The selection criteria included demonstrated compatibility with the program objectives (i.e., prior military experience, availability for a 40 day journey, and awareness of the need for a transformational intervention). The participants were connected as a group through a private Facebook site and interacted with one another for several months prior to the pilgrimage. Additionally, the participants were interviewed prior to beginning the pilgrimage, at one point during the pilgrimage and immediately following the conclusion of the pilgrimage journey. The interviews yielded themes related to loss, meaning construction, renewed hope in humanity, and a commitment to future goals. The lessons learned from this pilot project included a confirmation of the need for such a program, a need for greater focus on logistical details, and the recognition that the pilgrimage experience needs to continue in some manner once the veterans return home.

Keywords: pilgrimage, healing, military veterans, Camino de Santiago

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256 Antioxidant Potential of Sunflower Seed Cake Extract in Stabilization of Soybean Oil

Authors: Ivanor Zardo, Fernanda Walper Da Cunha, Júlia Sarkis, Ligia Damasceno Ferreira Marczak

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Lipid oxidation is one of the most important deteriorating processes in oil industry, resulting in the losses of nutritional value of oils as well as changes in color, flavor and other physiological properties. Autoxidation of lipids occurs naturally between molecular oxygen and the unsaturation of fatty acids, forming fat-free radicals, peroxide free radicals and hydroperoxides. In order to avoid the lipid oxidation in vegetable oils, synthetic antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tertiary butyl hydro-quinone (TBHQ) are commonly used. However, the use of synthetic antioxidants has been associated with several health side effects and toxicity. The use of natural antioxidants as stabilizers of vegetable oils is being suggested as a sustainable alternative to synthetic antioxidants. The alternative that has been studied is the use of natural extracts obtained mainly from fruits, vegetables and seeds, which have a well-known antioxidant activity related mainly to the presence of phenolic compounds. The sunflower seed cake is rich in phenolic compounds (1 4% of the total mass), being the chlorogenic acid the major constituent. The aim of this study was to evaluate the in vitro application of the phenolic extract obtained from the sunflower seed cake as a retarder of the lipid oxidation reaction in soybean oil and to compare the results with a synthetic antioxidant. For this, the soybean oil, provided from the industry without any addition of antioxidants, was subjected to an accelerated storage test for 17 days at 65 °C. Six samples with different treatments were submitted to the test: control sample, without any addition of antioxidants; 100 ppm of synthetic antioxidant BHT; mixture of 50 ppm of BHT and 50 ppm of phenolic compounds; and 100, 500 and 1200 ppm of phenolic compounds. The phenolic compounds concentration in the extract was expressed in gallic acid equivalents. To evaluate the oxidative changes of the samples, aliquots were collected after 0, 3, 6, 10 and 17 days and analyzed for the peroxide, diene and triene conjugate values. The soybean oil sample initially had a peroxide content of 2.01 ± 0.27 meq of oxygen/kg of oil. On the third day of the treatment, only the samples treated with 100, 500 and 1200 ppm of phenolic compounds showed a considerable oxidation retard compared to the control sample. On the sixth day of the treatment, the samples presented a considerable increase in the peroxide value (higher than 13.57 meq/kg), and the higher the concentration of phenolic compounds, the lower the peroxide value verified. From the tenth day on, the samples had a very high peroxide value (higher than 55.39 meq/kg), where only the sample containing 1200 ppm of phenolic compounds presented significant oxidation retard. The samples containing the phenolic extract were more efficient to avoid the formation of the primary oxidation products, indicating effectiveness to retard the reaction. Similar results were observed for dienes and trienes. Based on the results, phenolic compounds, especially chlorogenic acid (the major phenolic compound of sunflower seed cake), can be considered as a potential partial or even total substitute for synthetic antioxidants.

Keywords: chlorogenic acid, natural antioxidant, vegetables oil deterioration, waste valorization

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255 Single Cell Rna Sequencing Operating from Benchside to Bedside: An Interesting Entry into Translational Genomics

Authors: Leo Nnamdi Ozurumba-Dwight

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Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.

Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq

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254 Isolation and Structural Elucidation of 20 Hydroxyecdystone from Vitex doniana Sweet Stem Bark

Authors: Mustapha A. Tijjani, Fanna I. Abdulrahman, Irfan Z. Khan, Umar K. Sandabe, Cong Li

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Air dried sample V. doniana after collection and identification was extracted with ethanol and further partition with chloroform, ethyl acetate and n-butanol. Ethanolic extract (11.9g) was fractionated on a silica gel accelerated column chromatography using solvents such as n-hexane, ethyl acetate and methanol. Each eluent fractions (150ml aliquots) were collected and monitored with thin layer chromatography. Fractions with similar Rf values from same solvents system were pooled together. Phytochemical test of all the fractions were performed using standard procedure. Complete elution yielded 48 fractions (150ml/fraction) which were pooled to 24 fractions base on the Rf values. It was further recombined and 12 fractions were obtained on the basis on Rf values and coded Vd1 to Vd12 fractions. Vd8 was further eluted with ethylacetate and methanol and gave fourteen sub fractions Vd8-a, -Vd8-m. Fraction Vd8-a (56mg) gave a white crystal compound coded V1. It was further checked on TLC and observed under ultraviolet lamp and was found to give a single spot. The Rf values were calculated to be 0.433. The melting point was determined using Gallenkamp capillary melting point apparatus and found to be 241-243°C uncorrected. Characterization of the isolated compound coded V1 was done using FT-infra-red spectroscopy, HNMR, 13CNMR(1and 2D) and HRESI-MS. The IR spectrum of compound V1 shows prominent peaks that corresponds to OHstr (3365cm-1) and C=0 (1652cm-1) etc. This spectrum suggests that among the functional moiety in compound V1 are the carbonyl and hydroxyl group. The 1H NMR (400 MHz) spectrum of compound V1 in DMSO-d6 displayed five singlet signals at δ 0.72 (3H, s, H-18), 0.79 (3H, s, H-19), 1.03 (3H, s, H-21), 1.04 (3H, s, H-26), 1.06 (3H, s, H-27) each integrating for three protons indicating the five methyl functional groups present in the compound. It further showed a broad singlet at δ 5.58 integrated for 1 H due to an olefinic H-atom adjacent to the carbonyl carbon atom. Three signals at δ 3.10 (d, J = 9.0 Hz, H-22), 3.59 (m, 1H, 2H-a) and 3.72 (m, 1H, 3H-e), each integrating for one proton is due to oxymethine protons indicating that three oxymethine H-atoms are present in the compound. These all signals are characteristic to the ecdysteroid skeletons. The 13C-NMR spectrum showed the presence of 27 carbon atoms, suggesting that may be steroid skeleton. The DEPT-135 experiment showed the presence of five CH3, eight CH2, and seven CH groups, and seven quaternary C-atoms. The molecular formula was established as C27H44O7 by high resolution electron spray ionization-mass spectroscopy (HRESI-MS) positive ion mode m/z 481.3179. The signals in mass spectrum are 463, 445, and 427 peaks corresponding to losses of one, two, three, or four water molecules characteristic for ecdysterone skeleton reported in the literature. Based on the spectral analysis (HNMR, 13CNMR, DEPT, HMQC, IR, HRESI-MS) the compound V1 is thus concluded to have ecdysteriod skeleton and conclusively conforms with 2β, 3β 14α, 20R, 22R, 25-hexahydroxy-5 β cholest-7-ene-6- one, or 2, 3, 14, 20, 22, 25 hexahydroxy cholest-7-ene-6-one commonly known as 20-hydroxyecdysone.

Keywords: vitex, phytochemical, purification, isolation, chromatography, spectroscopy

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253 Investigating Seasonal Changes of Urban Land Cover with High Spatio-Temporal Resolution Satellite Data via Image Fusion

Authors: Hantian Wu, Bo Huang, Yuan Zeng

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Divisions between wealthy and poor, private and public landscapes are propagated by the increasing economic inequality of cities. While these are the spatial reflections of larger social issues and problems, urban design can at least employ spatial techniques that promote more inclusive rather than exclusive, overlapping rather than segregated, interlinked rather than disconnected landscapes. Indeed, the type of edge or border between urban landscapes plays a critical role in the way the environment is perceived. China experiences rapid urbanization, which poses unpredictable environmental challenges. The urban green cover and water body are under changes, which highly relevant to resident wealth and happiness. However, very limited knowledge and data on their rapid changes are available. In this regard, enhancing the monitoring of urban landscape with high-frequency method, evaluating and estimating the impacts of the urban landscape changes, and understating the driving forces of urban landscape changes can be a significant contribution for urban planning and studying. High-resolution remote sensing data has been widely applied to urban management in China. The map of urban land use map for the entire China of 2018 with 10 meters resolution has been published. However, this research focuses on the large-scale and high-resolution remote sensing land use but does not precisely focus on the seasonal change of urban covers. High-resolution remote sensing data has a long-operation cycle (e.g., Landsat 8 required 16 days for the same location), which is unable to satisfy the requirement of monitoring urban-landscape changes. On the other hand, aerial-remote or unmanned aerial vehicle (UAV) sensing are limited by the aviation-regulation and cost was hardly widely applied in the mega-cities. Moreover, those data are limited by the climate and weather conditions (e.g., cloud, fog), and those problems make capturing spatial and temporal dynamics is always a challenge for the remote sensing community. Particularly, during the rainy season, no data are available even for Sentinel Satellite data with 5 days interval. Many natural events and/or human activities drive the changes of urban covers. In this case, enhancing the monitoring of urban landscape with high-frequency method, evaluating and estimating the impacts of the urban landscape changes, and understanding the mechanism of urban landscape changes can be a significant contribution for urban planning and studying. This project aims to use the high spatiotemporal fusion of remote sensing data to create short-cycle, high-resolution remote sensing data sets for exploring the high-frequently urban cover changes. This research will enhance the long-term monitoring applicability of high spatiotemporal fusion of remote sensing data for the urban landscape for optimizing the urban management of landscape border to promoting the inclusive of the urban landscape to all communities.

Keywords: urban land cover changes, remote sensing, high spatiotemporal fusion, urban management

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252 Exploring the Neural Correlates of Different Interaction Types: A Hyperscanning Investigation Using the Pattern Game

Authors: Beata Spilakova, Daniel J. Shaw, Radek Marecek, Milan Brazdil

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Hyperscanning affords a unique insight into the brain dynamics underlying human interaction by simultaneously scanning two or more individuals’ brain responses while they engage in dyadic exchange. This provides an opportunity to observe dynamic brain activations in all individuals participating in interaction, and possible interbrain effects among them. The present research aims to provide an experimental paradigm for hyperscanning research capable of delineating among different forms of interaction. Specifically, the goal was to distinguish between two dimensions: (1) interaction structure (concurrent vs. turn-based) and (2) goal structure (competition vs cooperation). Dual-fMRI was used to scan 22 pairs of participants - each pair matched on gender, age, education and handedness - as they played the Pattern Game. In this simple interactive task, one player attempts to recreate a pattern of tokens while the second player must either help (cooperation) or prevent the first achieving the pattern (competition). Each pair played the game iteratively, alternating their roles every round. The game was played in two consecutive sessions: first the players took sequential turns (turn-based), but in the second session they placed their tokens concurrently (concurrent). Conventional general linear model (GLM) analyses revealed activations throughout a diffuse collection of brain regions: The cooperative condition engaged medial prefrontal cortex (mPFC) and posterior cingulate cortex (PCC); in the competitive condition, significant activations were observed in frontal and prefrontal areas, insula cortices and the thalamus. Comparisons between the turn-based and concurrent conditions revealed greater precuneus engagement in the former. Interestingly, mPFC, PCC and insulae are linked repeatedly to social cognitive processes. Similarly, the thalamus is often associated with a cognitive empathy, thus its activation may reflect the need to predict the opponent’s upcoming moves. Frontal and prefrontal activation most likely represent the higher attentional and executive demands of the concurrent condition, whereby subjects must simultaneously observe their co-player and place his own tokens accordingly. The activation of precuneus in the turn-based condition may be linked to self-other distinction processes. Finally, by performing intra-pair correlations of brain responses we demonstrate condition-specific patterns of brain-to-brain coupling in mPFC and PCC. Moreover, the degree of synchronicity in these neural signals related to performance on the game. The present results, then, show that different types of interaction recruit different brain systems implicated in social cognition, and the degree of inter-player synchrony within these brain systems is related to nature of the social interaction.

Keywords: brain-to-brain coupling, hyperscanning, pattern game, social interaction

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251 The Latest Salt Caravans: The Chinese Presence between Danakil and Tigray: Interdisciplinary Study to Integrate Chinese and African Relations in Ethiopia: Analyzing Road Evolution and Ethnographic Contexts

Authors: Erika Mattio

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The aim of this project is to study the Chinese presence in Ethiopia, in the area between the Saba River and the Coptic areas of the Tigray, with detailed documentation of the Danakil region, from which the salt pickers caravans departed; the study was created to understand the relationships and consequences of the Chinese advance in these areas, inhabited by tribes linked to ancient, still practiced religious rituals, and home to unique landscapes and archaeological sites. Official estimates of the number of Chinese in Africa vary widely; on the continent, there are increasingly diverse groups of Chinese migrants in terms of language, dialect, class, education, and employment. Based on this and on a very general state of the art, it was decided to increase the studies on this phenomenon, focusing the attention on one of the most interesting countries for its diversity, cultural wealth, and for strong Chinese presence: Ethiopia. The study will be integrated with interdisciplinary investigation methods, such as landscape archeology, historiographic research, participatory anthropology, geopolitics, and cultural anthropology and ethnology. There are two main objectives of the research. The first is to predict what will happen to these populations and how the territory will be modified, trying to monitor the growth of infrastructure in the country and the effects it will have on the population. Risk analyzes will be carried out to understand what the foreign presence may entail, such as the absence of sustenance for local populations, the ghettoization of foreigners, unemployment of natives and the exodus of the population to the capital; the relationships between families and the local population will be analyzed, trying to understand the dynamics of socialization and interaction. Thanks to the use of GIS, the areas affected by the Chinese presence will be geo-referenced and mapped, delimiting the areas most affected and creating a risk analysis, both in desert areas and in archaeologically and historically relevant areas. The second point is to document the life and rituals of Ethiopian populations in order not to lose the aspects of uniqueness that risk being lost. Local interviews will collect impressions and criticisms from the local population to understand if the Chinese presence is perceived as a threat or as a solution. Furthermore, Afar leaders in the Logya area will be interviewed, in truly exclusive research, to understand their links with the foreign presence. From the north, along the Saba river, we will move to the northwest, in the Tigray region, to know the impressions in the Coptic area, currently less threatened by the Chinese presence but still affected by urbanization proposals. There will also be documented the Coptic rituals of Gennà and Timkat, unique expressions of a millennial tradition. This will allow the understanding of whether the Maoist presence could influence the religious rites and forms of belief present in the country, or the country will maintain its cultural independence.

Keywords: Ethiopia, GIS, risk perceptions, salt caravans

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250 Research Project on Learning Rationality in Strategic Behaviors: Interdisciplinary Educational Activities in Italian High Schools

Authors: Giovanna Bimonte, Luigi Senatore, Francesco Saverio Tortoriello, Ilaria Veronesi

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The education process considers capabilities not only to be seen as a means to a certain end but rather as an effective purpose. Sen's capability approach challenges human capital theory, which sees education as an ordinary investment undertaken by individuals. A complex reality requires complex thinking capable of interpreting the dynamics of society's changes to be able to make decisions that can be rational for private, ethical and social contexts. Education is not something removed from the cultural and social context; it exists and is structured within it. In Italy, the "Mathematical High School Project" is a didactic research project is based on additional laboratory courses in extracurricular hours where mathematics intends to bring itself in a dialectical relationship with other disciplines as a cultural bridge between the two cultures, the humanistic and the scientific ones, with interdisciplinary educational modules on themes of strong impact in younger life. This interdisciplinary mathematics presents topics related to the most advanced technologies and contemporary socio-economic frameworks to demonstrate how mathematics is not only a key to reading but also a key to resolving complex problems. The recent developments in mathematics provide the potential for profound and highly beneficial changes in mathematics education at all levels, such as in socio-economic decisions. The research project is built to investigate whether repeated interactions can successfully promote cooperation among students as rational choice and if the skill, the context and the school background can influence the strategies choice and the rationality. A Laboratory on Game Theory as mathematical theory was conducted in the 4th year of the Mathematical High Schools and in an ordinary scientific high school of the Scientific degree program. Students played two simultaneous games of repeated Prisoner's Dilemma with an indefinite horizon, with two different competitors in each round; even though the competitors in each round will remain the same for the duration of the game. The results highlight that most of the students in the two classes used the two games with an immunization strategy against the risk of losing: in one of the games, they started by playing Cooperate, and in the other by the strategy of Compete. In the literature, theoretical models and experiments show that in the case of repeated interactions with the same adversary, the optimal cooperation strategy can be achieved by tit-for-tat mechanisms. In higher education, individual capacities cannot be examined independently, as conceptual framework presupposes a social construction of individuals interacting and competing, making individual and collective choices. The paper will outline all the results of the experimentation and the future development of the research.

Keywords: game theory, interdisciplinarity, mathematics education, mathematical high school

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249 Transient Heat Transfer: Experimental Investigation near the Critical Point

Authors: Andreas Kohlhepp, Gerrit Schatte, Wieland Christoph, Spliethoff Hartmut

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In recent years the research of heat transfer phenomena of water and other working fluids near the critical point experiences a growing interest for power engineering applications. To match the highly volatile characteristics of renewable energies, conventional power plants need to shift towards flexible operation. This requires speeding up the load change dynamics of steam generators and their heating surfaces near the critical point. In dynamic load transients, both a high heat flux with an unfavorable ratio to the mass flux and a high difference in fluid and wall temperatures, may cause problems. It may lead to deteriorated heat transfer (at supercritical pressures), dry-out or departure from nucleate boiling (at subcritical pressures), all cases leading to an extensive rise of temperatures. For relevant technical applications, the heat transfer coefficients need to be predicted correctly in case of transient scenarios to prevent damage to the heated surfaces (membrane walls, tube bundles or fuel rods). In transient processes, the state of the art method of calculating the heat transfer coefficients is using a multitude of different steady-state correlations for the momentarily existing local parameters for each time step. This approach does not necessarily reflect the different cases that may lead to a significant variation of the heat transfer coefficients and shows gaps in the individual ranges of validity. An algorithm was implemented to calculate the transient behavior of steam generators during load changes. It is used to assess existing correlations for transient heat transfer calculations. It is also desirable to validate the calculation using experimental data. By the use of a new full-scale supercritical thermo-hydraulic test rig, experimental data is obtained to describe the transient phenomena under dynamic boundary conditions as mentioned above and to serve for validation of transient steam generator calculations. Aiming to improve correlations for the prediction of the onset of deteriorated heat transfer in both, stationary and transient cases the test rig was specially designed for this task. It is a closed loop design with a directly electrically heated evaporation tube, the total heating power of the evaporator tube and the preheater is 1MW. To allow a big range of parameters, including supercritical pressures, the maximum pressure rating is 380 bar. The measurements contain the most important extrinsic thermo-hydraulic parameters. Moreover, a high geometric resolution allows to accurately predict the local heat transfer coefficients and fluid enthalpies.

Keywords: departure from nucleate boiling, deteriorated heat transfer, dryout, supercritical working fluid, transient operation of steam generators

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248 Mathematical Modelling of Bacterial Growth in Products of Animal Origin in Storage and Transport: Effects of Temperature, Use of Bacteriocins and pH Level

Authors: Benjamin Castillo, Luis Pastenes, Fernando Cordova

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The pathogen growth in animal source foods is a common problem in the food industry, causing monetary losses due to the spoiling of products or food intoxication outbreaks in the community. In this sense, the quality of the product is reflected by the population of deteriorating agents present in it, which are mainly bacteria. The factors which are likely associated with freshness in animal source foods are temperature and processing, storage, and transport times. However, the level of deterioration of products depends, in turn, on the characteristics of the bacterial population, causing the decomposition or spoiling, such as pH level and toxins. Knowing the growth dynamics of the agents that are involved in product contamination allows the monitoring for more efficient processing. This means better quality and reasonable costs, along with a better estimation of necessary time and temperature intervals for transport and storage in order to preserve product quality. The objective of this project is to design a secondary model that allows measuring the impact on temperature bacterial growth and the competition for pH adequacy and release of bacteriocins in order to describe such phenomenon and, thus, estimate food product half-life with the least possible risk of deterioration or spoiling. In order to achieve this objective, the authors propose an analysis of a three-dimensional ordinary differential which includes; logistic bacterial growth extended by the inhibitory action of bacteriocins including the effect of the medium pH; change in the medium pH levels through an adaptation of the Luedeking-Piret kinetic model; Bacteriocin concentration modeled similarly to pH levels. These three dimensions are being influenced by the temperature at all times. Then, this differential system is expanded, taking into consideration the variable temperature and the concentration of pulsed bacteriocins, which represent characteristics inherent of the modeling, such as transport and storage, as well as the incorporation of substances that inhibit bacterial growth. The main results lead to the fact that temperature changes in an early stage of transport increased the bacterial population significantly more than if it had increased during the final stage. On the other hand, the incorporation of bacteriocins, as in other investigations, proved to be efficient in the short and medium-term since, although the population of bacteria decreased, once the bacteriocins were depleted or degraded over time, the bacteria eventually returned to their regular growth rate. The efficacy of the bacteriocins at low temperatures decreased slightly, which equates with the fact that their natural degradation rate also decreased. In summary, the implementation of the mathematical model allowed the simulation of a set of possible bacteria present in animal based products, along with their properties, in various transport and storage situations, which led us to state that for inhibiting bacterial growth, the optimum is complementary low constant temperatures and the initial use of bacteriocins.

Keywords: bacterial growth, bacteriocins, mathematical modelling, temperature

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247 Decomposition of the Discount Function Into Impatience and Uncertainty Aversion. How Neurofinance Can Help to Understand Behavioral Anomalies

Authors: Roberta Martino, Viviana Ventre

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Intertemporal choices are choices under conditions of uncertainty in which the consequences are distributed over time. The Discounted Utility Model is the essential reference for describing the individual in the context of intertemporal choice. The model is based on the idea that the individual selects the alternative with the highest utility, which is calculated by multiplying the cardinal utility of the outcome, as if the reception were instantaneous, by the discount function that determines a decrease in the utility value according to how the actual reception of the outcome is far away from the moment the choice is made. Initially, the discount function was assumed to have an exponential trend, whose decrease over time is constant, in line with a profile of a rational investor described by classical economics. Instead, empirical evidence called for the formulation of alternative, hyperbolic models that better represented the actual actions of the investor. Attitudes that do not comply with the principles of classical rationality are termed anomalous, i.e., difficult to rationalize and describe through normative models. The development of behavioral finance, which describes investor behavior through cognitive psychology, has shown that deviations from rationality are due to the limited rationality condition of human beings. What this means is that when a choice is made in a very difficult and information-rich environment, the brain does a compromise job between the cognitive effort required and the selection of an alternative. Moreover, the evaluation and selection phase of the alternative, the collection and processing of information, are dynamics conditioned by systematic distortions of the decision-making process that are the behavioral biases involving the individual's emotional and cognitive system. In this paper we present an original decomposition of the discount function to investigate the psychological principles of hyperbolic discounting. It is possible to decompose the curve into two components: the first component is responsible for the smaller decrease in the outcome as time increases and is related to the individual's impatience; the second component relates to the change in the direction of the tangent vector to the curve and indicates how much the individual perceives the indeterminacy of the future indicating his or her aversion to uncertainty. This decomposition allows interesting conclusions to be drawn with respect to the concept of impatience and the emotional drives involved in decision-making. The contribution that neuroscience can make to decision theory and inter-temporal choice theory is vast as it would allow the description of the decision-making process as the relationship between the individual's emotional and cognitive factors. Neurofinance is a discipline that uses a multidisciplinary approach to investigate how the brain influences decision-making. Indeed, considering that the decision-making process is linked to the activity of the prefrontal cortex and amygdala, neurofinance can help determine the extent to which abnormal attitudes respect the principles of rationality.

Keywords: impatience, intertemporal choice, neurofinance, rationality, uncertainty

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246 Intriguing Modulations in the Excited State Intramolecular Proton Transfer Process of Chrysazine Governed by Host-Guest Interactions with Macrocyclic Molecules

Authors: Poojan Gharat, Haridas Pal, Sharmistha Dutta Choudhury

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Tuning photophysical properties of guest dyes through host-guest interactions involving macrocyclic hosts are the attractive research areas since past few decades, as these changes can directly be implemented in chemical sensing, molecular recognition, fluorescence imaging and dye laser applications. Excited state intramolecular proton transfer (ESIPT) is an intramolecular prototautomerization process display by some specific dyes. The process is quite amenable to tunability by the presence of different macrocyclic hosts. The present study explores the interesting effect of p-sulfonatocalix[n]arene (SCXn) and cyclodextrin (CD) hosts on the excited-state prototautomeric equilibrium of Chrysazine (CZ), a model antitumour drug. CZ exists exclusively in its normal form (N) in the ground state. However, in the excited state, the excited N* form undergoes ESIPT along with its pre-existing intramolecular hydrogen bonds, giving the excited state prototautomer (T*). Accordingly, CZ shows a single absorption band due to N form, but two emission bands due to N* and T* forms. Facile prototautomerization of CZ is considerably inhibited when the dye gets bound to SCXn hosts. However, in spite of lower binding affinity, the inhibition is more profound with SCX6 host as compared to SCX4 host. For CD-CZ system, while prototautomerization process is hindered by the presence of β-CD, it remains unaffected in the presence of γCD. Reduction in the prototautomerization process of CZ by SCXn and βCD hosts is unusual, because T* form is less dipolar in nature than the N*, hence binding of CZ within relatively hydrophobic hosts cavities should have enhanced the prototautomerization process. At the same time, considering the similar chemical nature of two CD hosts, their effect on prototautomerization process of CZ would have also been similar. The atypical effects on the prototautomerization process of CZ by the studied hosts are suggested to arise due to the partial inclusion or external binding of CZ with the hosts. As a result, there is a strong possibility of intermolecular H-bonding interaction between CZ dye and the functional groups present at the portals of SCXn and βCD hosts. Formation of these intermolecular H-bonds effectively causes the pre-existing intramolecular H-bonding network within CZ molecule to become weak, and this consequently reduces the prototautomerization process for the dye. Our results suggest that rather than the binding affinity between the dye and host, it is the orientation of CZ in the case of SCXn-CZ complexes and the binding stoichiometry in the case of CD-CZ complexes that play the predominant role in influencing the prototautomeric equilibrium of the dye CZ. In the case of SCXn-CZ complexes, the results obtained through experimental findings are well supported by quantum chemical calculations. Similarly for CD-CZ systems, binding stoichiometries obtained through geometry optimization studies on the complexes between CZ and CD hosts correlate nicely with the experimental results. Formation of βCD-CZ complexes with 1:1 stoichiometry while formation of γCD-CZ complexes with 1:1, 1:2 and 2:2 stoichiometries are revealed from geometry optimization studies and these results are in good accordance with the observed effects by the βCD and γCD hosts on the ESIPT process of CZ dye.

Keywords: intermolecular proton transfer, macrocyclic hosts, quantum chemical studies, photophysical studies

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245 Newly Designed Ecological Task to Assess Cognitive Map Reading Ability: Behavioral Neuro-Anatomic Correlates of Mental Navigation

Authors: Igor Faulmann, Arnaud Saj, Roland Maurer

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Spatial cognition consists in a plethora of high level cognitive abilities: among them, the ability to learn and to navigate in large scale environments is probably one of the most complex skills. Navigation is thought to rely on the ability to read a cognitive map, defined as an allocentric representation of ones environment. Those representations are of course intimately related to the two geometrical primitives of the environment: distance and direction. Also, many recent studies point to a predominant hippocampal and para-hippocampal role in spatial cognition, as well as in the more specific cluster of navigational skills. In a previous study in humans, we used a newly validated test assessing cognitive map processing by evaluating the ability to judge relative distances and directions: the CMRT (Cognitive Map Recall Test). This study identified in topographically disorientated patients (1) behavioral differences between the evaluation of distances and of directions, and (2) distinct causality patterns assessed via VLSM (i.e., distinct cerebral lesions cause distinct response patterns depending on the modality (distance vs direction questions). Thus, we hypothesized that: (1) if the CMRT really taps into the same resources as real navigation, there would be hippocampal, parahippocampal, and parietal activation, and (2) there exists underlying neuroanatomical and functional differences between the processing of this two modalities. Aiming toward a better understanding of the neuroanatomical correlates of the CMRT in humans, and more generally toward a better understanding of how the brain processes the cognitive map, we adapted the CMRT as an fMRI procedure. 23 healthy subjects (11 women, 12 men), all living in Geneva for at least 2 years, underwent the CMRT in fMRI. Results show, for distance and direction taken together, than the most active brain regions are the parietal, frontal and cerebellar parts. Additionally, and as expected, patterns of brain activation differ when comparing the two modalities. Furthermore, distance processing seems to rely more on parietal regions (compared to other brain regions in the same modality and also to direction). It is interesting to notice that no significant activity was observed in the hippocampal or parahippocampal areas. Direction processing seems to tap more into frontal and cerebellar brain regions (compared to other brain regions in the same modality and also to distance). Significant hippocampal and parahippocampal activity has been shown only in this modality. This results demonstrated a complex interaction of structures which are compatible with response patterns observed in other navigational tasks, thus showing that the CMRT taps at least partially into the same brain resources as real navigation. Additionally, differences between the processing of distances and directions leads to the conclusion that the human brain processes each modality distinctly. Further research should focus on the dynamics of this processing, allowing a clearer understanding between the two sub-processes.

Keywords: cognitive map, navigation, fMRI, spatial cognition

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244 Phenotypic and Molecular Heterogeneity Linked to the Magnesium Transporter CNNM2

Authors: Reham Khalaf-Nazzal, Imad Dweikat, Paula Gimenez, Iker Oyenarte, Alfonso Martinez-Cruz, Domonik Muller

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Metal cation transport mediator (CNNM) gene family comprises 4 isoforms that are expressed in various human tissues. Structurally, CNNMs are complex proteins that contain an extracellular N-terminal domain preceding a DUF21 transmembrane domain, a ‘Bateman module’ and a C-terminal cNMP-binding domain. Mutations in CNNM2 cause familial dominant hypomagnesaemia. Growing evidence highlights the role of CNNM2 in neurodevelopment. Mutations in CNNM2 have been implicated in epilepsy, intellectual disability, schizophrenia, and others. In the present study, we aim to elucidate the function of CNNM2 in the developing brain. Thus, we present the genetic origin of symptoms in two family cohorts. In the first family, three siblings of a consanguineous Palestinian family in which parents are first cousins, and consanguinity ran over several generations, presented a varying degree of intellectual disability, cone-rod dystrophy, and autism spectrum disorder. Exome sequencing and segregation analysis revealed the presence of homozygous pathogenic mutation in the CNNM2 gene, the parents were heterozygous for that gene mutation. Magnesium blood levels were normal in the three children and their parents in several measurements. They had no symptoms of hypomagnesemia. The CNNM2 mutation in this family was found to locate in the CBS1 domain of the CNNM2 protein. The crystal structure of the mutated CNNM2 protein was not significantly different from the wild-type protein, and the binding of AMP or MgATP was not dramatically affected. This suggests that the CBS1 domain could be involved in pure neurodevelopmental functions independent of its magnesium-handling role, and this mutation could have affected a protein partner binding or other functions in this protein. In the second family, another autosomal dominant CNNM2 mutation was found to run in a large family with multiple individuals over three generations. All affected family members had hypomagnesemia and hypermagnesuria. Oral supplementation of magnesium did not increase the levels of magnesium in serum significantly. Some affected members of this family have defects in fine motor skills such as dyslexia and dyslalia. The detected mutation is located in the N-terminal part, which contains a signal peptide thought to be involved in the sorting and routing of the protein. In this project, we describe heterogenous clinical phenotypes related to CNNM2 mutations and protein functions. In the first family, and up to the authors’ knowledge, we report for the first time the involvement of CNNM2 in retinal photoreceptor development and function. In addition, we report the presence of a neurophenotype independent of magnesium status related to the CNNM2 protein mutation. Taking into account the different modes of inheritance and the different positions of the mutations within CNNM2 and its different structural and functional domains, it is likely that CNNM2 might be involved in a wide spectrum of neuropsychiatric comorbidities with considerable varying phenotypes.

Keywords: magnesium transport, autosomal recessive, autism, neurodevelopment, CBS domain

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243 Two-wavelength High-energy Cr:LiCaAlF6 MOPA Laser System for Medical Multispectral Optoacoustic Tomography

Authors: Radik D. Aglyamov, Alexander K. Naumov, Alexey A. Shavelev, Oleg A. Morozov, Arsenij D. Shishkin, Yury P.Brodnikovsky, Alexander A.Karabutov, Alexander A. Oraevsky, Vadim V. Semashko

Abstract:

The development of medical optoacoustic tomography with the using human blood as endogenic contrast agent is constrained by the lack of reliable, easy-to-use and inexpensive sources of high-power pulsed laser radiation in the spectral region of 750-900 nm [1-2]. Currently used titanium-sapphire, alexandrite lasers or optical parametric light oscillators do not provide the required and stable output characteristics, they are structurally complex, and their cost is up to half the price of diagnostic optoacoustic systems. Here we are developing the lasers based on Cr:LiCaAlF6 crystals which are free of abovementioned disadvantages and provides intensive ten’s ns-range tunable laser radiation at specific absorption bands of oxy- (~840 nm) and -deoxyhemoglobin (~757 nm) in the blood. Cr:LiCAF (с=3 at.%) crystals were grown in Kazan Federal University by the vertical directional crystallization (Bridgman technique) in graphite crucibles in a fluorinating atmosphere at argon overpressure (P=1500 hPa) [3]. The laser elements have cylinder shape with the diameter of 8 mm and 90 mm in length. The direction of the optical axis of the crystal was normal to the cylinder generatrix, which provides the π-polarized laser action correspondent to maximal stimulated emission cross-section. The flat working surfaces of the active elements were polished and parallel to each other with an error less than 10”. No any antireflection coating was applied. The Q-switched master oscillator-power amplifiers laser system (MOPA) with the dual-Xenon flashlamp pumping scheme in diffuse-reflectivity close-coupled head were realized. A specially designed laser cavity, consisting of dielectric highly reflective reflectors with a 2 m-curvature radius, a flat output mirror, a polarizer and Q-switch sell, makes it possible to operate sequentially in a circle (50 ns - laser one pulse after another) at wavelengths of 757 and 840 nm. The programmable pumping system from Tomowave Laser LLC (Russia) provided independent to each pulses (up to 250 J at 180 μs) pumping to equalize the laser radiation intensity at these wavelengths. The MOPA laser operates at 10 Hz pulse repetition rate with the output energy up to 210 mJ. Taking into account the limitations associated with physiological movements and other characteristics of patient tissues, the duration of laser pulses and their energy allows molecular and functional high-contrast imaging to depths of 5-6 cm with a spatial resolution of at least 1 mm. Highly likely the further comprehensive design of laser allows improving the output properties and realizing better spatial resolution of medical multispectral optoacoustic tomography systems.

Keywords: medical optoacoustic, endogenic contrast agent, multiwavelength tunable pulse lasers, MOPA laser system

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242 Anti-proliferative Activity and HER2 Receptor Expression Analysis of MCF-7 (Breast Cancer Cell) Cells by Plant Extract Coleus Barbatus (Andrew)

Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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241 One Pot Synthesis of Cu–Ni–S/Ni Foam for the Simultaneous Removal and Detection of Norfloxacin

Authors: Xincheng Jiang, Yanyan An, Yaoyao Huang, Wei Ding, Manli Sun, Hong Li, Huaili Zheng

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The residual antibiotics in the environment will pose a threat to the environment and human health. Thus, efficient removal and rapid detection of norfloxacin (NOR) in wastewater is very important. The main sources of NOR pollution are the agricultural, pharmaceutical industry and hospital wastewater. The total consumption of NOR in China can reach 5440 tons per year. It is found that neither animals nor humans can totally absorb and metabolize NOR, resulting in the excretion of NOR into the environment. Therefore, residual NOR has been detected in water bodies. The hazards of NOR in wastewater lie in three aspects: (1) the removal capacity of the wastewater treatment plant for NOR is limited (it is reported that the average removal efficiency of NOR in the wastewater treatment plant is only 68%); (2) NOR entering the environment will lead to the emergence of drug-resistant strains; (3) NOR is toxic to many aquatic species. At present, the removal and detection technologies of NOR are applied separately, which leads to a cumbersome operation process. The development of simultaneous adsorption-flocculation removal and FTIR detection of pollutants has three advantages: (1) Adsorption-flocculation technology promotes the detection technology (the enrichment effect on the material surface improves the detection ability); (2) The integration of adsorption-flocculation technology and detection technology reduces the material cost and makes the operation easier; (3) FTIR detection technology endows the water treatment agent with the ability of molecular recognition and semi-quantitative detection for pollutants. Thus, it is of great significance to develop a smart water treatment material with high removal capacity and detection ability for pollutants. This study explored the feasibility of combining NOR removal method with the semi-quantitative detection method. A magnetic Cu-Ni-S/Ni foam was synthesized by in-situ loading Cu-Ni-S nanostructures on the surface of Ni foam. The novelty of this material is the combination of adsorption-flocculation technology and semi-quantitative detection technology. Batch experiments showed that Cu-Ni-S/Ni foam has a high removal rate of NOR (96.92%), wide pH adaptability (pH=4.0-10.0) and strong ion interference resistance (0.1-100 mmol/L). According to the Langmuir fitting model, the removal capacity can reach 417.4 mg/g at 25 °C, which is much higher than that of other water treatment agents reported in most studies. Characterization analysis indicated that the main removal mechanisms are surface complexation, cation bridging, electrostatic attraction, precipitation and flocculation. Transmission FTIR detection experiments showed that NOR on Cu-Ni-S/Ni foam has easily recognizable FTIR fingerprints; the intensity of characteristic peaks roughly reflects the concentration information to some extent. This semi-quantitative detection method has a wide linear range (5-100 mg/L) and a low limit of detection (4.6 mg/L). These results show that Cu-Ni-S/Ni foam has excellent removal performance and semi-quantitative detection ability of NOR molecules. This paper provides a new idea for designing and preparing multi-functional water treatment materials to achieve simultaneous removal and semi-quantitative detection of organic pollutants in water.

Keywords: adsorption-flocculation, antibiotics detection, Cu-Ni-S/Ni foam, norfloxacin

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240 Ramification of Pemphigus Vulgaris Sera and the Monoclonal Antibody Against Desmoglein-3 on Nrf2 Expression in Keratinocyte Cultures

Authors: Faris Mohsin Alabeedi

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Pemphigus Vulgaris (PV) is a life-threatening autoimmune blistering disease characterized by the presence of autoantibodies directed against the epidermis's surface proteins. There are two forms of PV, mucocutaneous and mucosal-dominant PV. Disruption of the cell junctions is a hallmark of PV due to the autoantibodies targeting the desmosomal cadherins, desmoglein-3 (Dsg3) and desmoglein-1, leading to acantholysis in the skin and mucous membrane. Although the pathogenesis of PV is known, the detailed molecular events remain not fully understood. Our recent study has shown that both the PV sera and pathogenic anti-Dsg3 antibody AK23 can induce ROS and cause oxidative stress in cultured keratinocytes. In line with our finding, other independent studies also demonstrate oxidative stress in PV. Since Nrf2 plays a crucial role in cellular anti-oxidative stress response, we hypothesize that the expression of Nrf2 may alter in PV. Thus, treatment of cells with PV sera or AK23 may cause changes in Nrf2 expression and distribution. The purpose of this study was to examine the effect of AK23 and PV sera on Nrf2 in a normal human keratinocyte cell line, such as NTERT cells. Both a time-course and dose-dependent experiments with AK23, alongside the matched isotype control IgG, were performed in keratinocyte cultures and analysed by immunofluorescence for Nrf2 and Dsg3. Additionally, the same approach was conducted with the sera from PV patients and healthy individuals that served as a control in this study. All the fluorescent images were analysed using ImageJ software. Each experiment was repeated twice. In general, variations were observed throughout this study. In the dose-response experiments, although enhanced Dsg3 expression was consistently detected in AK23 treated cells, the expression of Nrf2 showed no consistent findings between the experiments, although changes in its expression were noticeable in cells treated with AK23. In the time-course study, a trend with induction of Nrf2 over time was shown in control cells treated with mouse isotype IgG. Treatment with AK23 showed a reduction of Nrf2 in a time-dependent manner, especially at the 24-hour time point. However, the earlier time points, such as 2 hours and 6 hours with AK23 treatments, detected somewhat variations. Finally, PV sera caused a decrease of Dsg3, but on the other hand, variations were observed in Nrf2 expression in PV sera treated cells. In general, PV sera seemed to cause a reduction of Nrf2 in the majority of PV sera treated samples. In addition, more pronounced cytoplasmic expression of Nrf2 has been observed in PV sera treated cells than those treated with AK23, suggesting that polyclonal and monoclonal IgG might induce a different effect on Nrf2 expression and distribution. Further experimental studies are crucial to obtain a more coincide global view of Nrf2-mediated gene regulation. In particular, Pemphigus Voulgaris studies assessing how the Nrf2-dependent network changes from a physiological to a pathological condition can provide insight into disease mechanisms and perhaps initiate further treatment approaches.

Keywords: pemphigus vulgaris, monoclonal antibody against desmoglein-3, Nrf2 oxidative stress, keratinocyte cultures

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239 Impact of U.S. Insurance Reimbursement Policy on Healthcare Business and Entrepreneurship

Authors: Iris Xiaohong Quan, Sharon Qi, Kelly Tianqin Shi

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This study focuses on the critical role of insurance policies in a world grappling with increasing mental health challenges, as they significantly influence the dynamics of healthcare businesses and entrepreneurial ventures. The paper utilizes the mental health sector as a case to examine the impact of insurance policies on healthcare service providers, entrepreneurs, and individuals seeking mental health support. This paper addressed the following research questions: To what extent do changes in insurance reimbursement policies affect the accessibility and affordability of mental health services for patients, and how does this impact the overall demand for such services? What are the barriers and opportunities that mental health entrepreneurs face and what strategies and adaptations do mental health businesses employ when navigating the evolving landscape of insurance reimbursement policies? How do changes in insurance reimbursement policies, specifically related to mental health services, influence the financial viability and sustainability of mental health clinics and private practices? Employing a self-designed survey aimed at autism spectrum disorder (ASD) treatment companies, alongside two in-depth case studies and an analysis of pertinent insurance policies and documents, this research aims to elucidate the multifaceted influence of insurance policies on the mental health industry. The findings from this study reveal how insurance policies shape the landscape of mental health businesses and their operations. A total of 821 autism treatment organizations or offices were contacted by telephone between November 1, 2019, and January 31, 2020. About half of the offices (53.33%) were established in the past five years, and 80% were established in the past 15 years. There is a significant increase in the establishment of ABA service centers in the recent two decades as a result of autism insurance reform, the increasing social awareness of ASD, and the redefinition of autism. In addition, almost half of the ABA service providers we surveyed had a patient size ranging from 20 to 50 in the year when the residence state passed the legislation for autism insurance coverage. On average, an ABA service provider works with 5.3 insurance companies. This research find that insurance is the main source of revenue for most ABA service providers. However, our survey reveals that clients’ out of pocket payment has been the second main revenue sources. Despite the changes of regulations and insurance policies in all states, clients still have to pay a fraction of, if not all, the ABA treatment service fees out of pocket. This research shows that some ABA service providers seek federal and government funds and grants to support their services and businesses. Our further analysis with the in-depth case studies and other secondary data also indicate the rise of entrepreneurial startups in the mental health industry. Overall, this research sheds light on both the challenges and opportunities presented by insurance policies in the mental health sector, offering insights into the new industry landscape.

Keywords: entrepreneurship, healthcare policy, insurance policy, mental health industry

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238 The Dynamics of a Droplet Spreading on a Steel Surface

Authors: Evgeniya Orlova, Dmitriy Feoktistov, Geniy Kuznetsov

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Spreading of a droplet over a solid substrate is a key phenomenon observed in the following engineering applications: thin film coating, oil extraction, inkjet printing, and spray cooling of heated surfaces. Droplet cooling systems are known to be more effective than film or rivulet cooling systems. It is caused by the greater evaporation surface area of droplets compared with the film of the same mass and wetting surface. And the greater surface area of droplets is connected with the curvature of the interface. Location of the droplets on the cooling surface influences on the heat transfer conditions. The close distance between the droplets provides intensive heat removal, but there is a possibility of their coalescence in the liquid film. The long distance leads to overheating of the local areas of the cooling surface and the occurrence of thermal stresses. To control the location of droplets is possible by changing the roughness, structure and chemical composition of the surface. Thus, control of spreading can be implemented. The most important characteristic of spreading of droplets on solid surfaces is a dynamic contact angle, which is a function of the contact line speed or capillary number. However, there is currently no universal equation, which would describe the relationship between these parameters. This paper presents the results of the experimental studies of water droplet spreading on metal substrates with different surface roughness. The effect of the droplet growth rate and the surface roughness on spreading characteristics was studied at low capillary numbers. The shadow method using high speed video cameras recording up to 10,000 frames per seconds was implemented. A droplet profile was analyzed by Axisymmetric Drop Shape Analyses techniques. According to change of the dynamic contact angle and the contact line speed three sequential spreading stages were observed: rapid increase in the dynamic contact angle; monotonous decrease in the contact angle and the contact line speed; and form of the equilibrium contact angle at constant contact line. At low droplet growth rate, the dynamic contact angle of the droplet spreading on the surfaces with the maximum roughness is found to increase throughout the spreading time. It is due to the fact that the friction force on such surfaces is significantly greater than the inertia force; and the contact line is pinned on microasperities of a relief. At high droplet growth rate the contact angle decreases during the second stage even on the surfaces with the maximum roughness, as in this case, the liquid does not fill the microcavities, and the droplet moves over the “air cushion”, i.e. the interface is a liquid/gas/solid system. Also at such growth rates pulsation of liquid flow was detected; and the droplet oscillates during the spreading. Thus, obtained results allow to conclude that it is possible to control spreading by using the surface roughness and the growth rate of droplets on surfaces as varied factors. Also, the research findings may be used for analyzing heat transfer in rivulet and drop cooling systems of high energy equipment.

Keywords: contact line speed, droplet growth rate, dynamic contact angle, shadow system, spreading

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237 Long Non-Coding RNAs Mediated Regulation of Diabetes in Humanized Mouse

Authors: Md. M. Hossain, Regan Roat, Jenica Christopherson, Colette Free, Zhiguang Guo

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Long noncoding RNA (lncRNA) mediated post-transcriptional gene regulation, and their epigenetic landscapes have been shown to be involved in many human diseases. However, their regulation in diabetes through governing islet’s β-cell function and survival needs to be elucidated. Due to the technical and ethical constraints, it is difficult to study their role in β-cell function and survival in human under in vivo condition. In this study, humanized mice have been developed through transplanting human pancreatic islet under the kidney capsule of NOD.SCID mice and induced β-cell death leading to diabetes condition to study lncRNA mediated regulation. For this, human islets from 3 donors (3000 IEQ, purity > 80%) were transplanted under the kidney capsule of STZ induced diabetic NOD.scid mice. After at least 2 weeks of normoglycecemia, lymphocytes from diabetic NOD mice were adoptively transferred and islet grafts were collected once blood glucose reached > 200 mg/dl. RNA from human donor islets, islet grafts from humanized mice with either adoptive lymphocyte transfer (ALT) or PBS control (CTL) were ribodepleted; barcoded fragment libraries were constructed and sequenced on the Ion Proton sequencer. lncRNA expression in isolated human islets, islet grafts from humanized mice with and without induced β-cell death and their regulation in human islets function in vitro under glucose challenge, cytokine mediated inflammation and induced apoptotic condition were investigated. Out of 3155 detected lncRNAs, 299 that highly expressed in islets were found to be significantly downregulated and 224 upregulated in ALT compared to CTL. Most of these are found to be collocated within 5 kb upstream and 1 kb downstream of 788 up- and 624 down-regulated mRNAs. Genomic Regions Enrichment of Annotations Analysis revealed deregulated and collocated genes are related to pancreas endocrine development; insulin synthesis, processing, and secretion; pancreatitis and diabetes. Many of them, that found to be located within enhancer domains for islet specific gene activity, are associated to the deregulation of known islet/βcell specific transcription factors and genes that are important for β-cell differentiation, identity, and function. RNA sequencing analysis revealed aberrant lncRNA expression which is associated to the deregulated mRNAs in β-cell function as well as in molecular pathways related to diabetes. A distinct set of candidate lncRNA isoforms were identified as highly enriched and specific to human islets, which are deregulated in human islets from donors with different BMIs and with type 2 diabetes. These RNAs show an interesting regulation in cultured human islets under glucose stimulation and with induced β-cell death by cytokines. Aberrant expression of these lncRNAs was detected in the exosomes from the media of islets cultured with cytokines. Results of this study suggest that the islet specific lncRNAs are deregulated in human islet with β-cell death, hence important in diabetes. These lncRNAs might be important for human β-cell function and survival thus could be used as biomarkers and novel therapeutic targets for diabetes.

Keywords: β-cell, humanized mouse, pancreatic islet, LncRNAs

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236 Dragonflies (Odonata) Reflect Climate Warming Driven Changes in High Mountain Invertebrates Populations

Authors: Nikola Góral, Piotr Mikołajczuk, Paweł Buczyński

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Much scientific research in the last 20 years has focused on the influence of global warming on the distribution and phenology of living organisms. Three potential responses to climate change are predicted: individual species may become extinct, adapt to new conditions in their existing range or change their range by migrating to places where climatic conditions are more favourable. It means not only migration to areas in other latitudes, but also different altitudes. In the case of dragonflies (Odonata), monitoring in Western Europe has shown that in response to global warming, dragonflies tend to change their range to a more northern one. The strongest response to global warming is observed in arctic and alpine species, as well as in species capable of migrating over long distances. The aim of the research was to assess whether the fauna of aquatic insects in high-mountain habitats has changed as a result of climate change and, if so, how big and what type these changes are. Dragonflies were chosen as a model organism because of their fast reaction to changes in the environment: they have high migration abilities and short life cycle. The state of the populations of boreal-mountain species and the extent to which lowland species entered high altitudes was assessed. The research was carried out on 20 sites in Western Sudetes, Southern Poland. They were located at an altitude of between 850 and 1250 m. The selected sites were representative of many types of valuable alpine habitats (subalpine raised bog, transitional spring bog, habitats associated with rivers and mountain streams). Several sites of anthropogenic origin were also selected. Thanks to this selection, a wide characterization of the fauna of the Karkonosze was made and it was compared whether the studied processes proceeded differently, depending on whether the habitat is primary or secondary. Both imagines and larvae were examined (by taking hydrobiological samples with a kick-net), and exuviae were also collected. Individual species dragonflies were characterized in terms of their reproductive, territorial and foraging behaviour. During each inspection, the basic physicochemical parameters of the water were measured. The population of the high-mountain dragonfly Somatochlora alpestris turned out to be in a good condition. This species was noted at several sites. Some of those sites were situated relatively low (995 m AMSL), which proves that the thermal conditions at the lower altitudes might be still optimal for this species. The protected by polish law species Somatochlora arctica, Aeshna subarctica and Leucorrhinia albifrons, as well as strongly associated with bogs Leucorrhinia dubia and Aeshna juncea bogs were observed. However, they were more frequent and more numerous in habitats of anthropogenic origin, which may suggest minor changes in the habitat preferences of dragonflies. The subject requires further research and observations over a longer time scale.

Keywords: alpine species, bioindication, global warming, habitat preferences, population dynamics

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