Search results for: Gian Andrea Blengini

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

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

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

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Authors: Mylena McCoggle, Shyra Wilson, Andrea Rivera, Rocio Alba-Flores

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This work introduces the use of EMGs (electromyography) from muscle sensors to develop an Artificial Neural Network (ANN) for pattern recognition to control a small unmanned aerial vehicle. The objective of this endeavor exhibits interfacing drone applications beyond manual control directly. MyoWare Muscle sensor contains three EMG electrodes (dual and single type) used to collect signals from the posterior (extensor) and anterior (flexor) forearm and the bicep. Collection of raw voltages from each sensor were connected to an Arduino Uno and a data processing algorithm was developed with the purpose of interpreting the voltage signals given when performing flexing, resting, and motion of the arm. Each sensor collected eight values over a two-second period for the duration of one minute, per assessment. During each two-second interval, the movements were alternating between a resting reference class and an active motion class, resulting in controlling the motion of the drone with left and right movements. This paper further investigated adding up to three sensors to differentiate between hand gestures to control the principal motions of the drone (left, right, up, and land). The hand gestures chosen to execute these movements were: a resting position, a thumbs up, a hand swipe right motion, and a flexing position. The MATLAB software was utilized to collect, process, and analyze the signals from the sensors. The protocol (machine learning tool) was used to classify the hand gestures. To generate the input vector to the ANN, the mean, root means squared, and standard deviation was processed for every two-second interval of the hand gestures. The neuromuscular information was then trained using an artificial neural network with one hidden layer of 10 neurons to categorize the four targets, one for each hand gesture. Once the machine learning training was completed, the resulting network interpreted the processed inputs and returned the probabilities of each class. Based on the resultant probability of the application process, once an output was greater or equal to 80% of matching a specific target class, the drone would perform the motion expected. Afterward, each movement was sent from the computer to the drone through a Wi-Fi network connection. These procedures have been successfully tested and integrated into trial flights, where the drone has responded successfully in real-time to predefined command inputs with the machine learning algorithm through the MyoWare sensor interface. The full paper will describe in detail the database of the hand gestures, the details of the ANN architecture, and confusion matrices results.

Keywords: artificial neural network, biosensors, electromyography, machine learning, MyoWare muscle sensors, Arduino

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Authors: Eva Masson, Andrea Kübler

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Anxiety disorders (AD) are the most prevalent psychological disorders. However, far from most affected individuals are diagnosed and receive treatment. This gap is probably due to the diagnosis criteria, relying on symptoms (according to the DSM-5 definition) with no objective biomarker. Approach-avoidance conflict tasks are one common approach to simulate such disorders in a lab setting, with most of the paradigms focusing on the relationships between behavior and neurophysiology. Approach-avoidance conflict tasks typically place participants in a situation where they have to make a decision that leads to both positive and negative outcomes, thereby sending conflicting signals that trigger the Behavioral Inhibition System (BIS). Furthermore, behavioral validation of such paradigms adds credibility to the tasks – with overt conflict behavior, it is safer to assume that the task actually induced a conflict. Some of those tasks have linked asymmetrical frontal brain activity to induced conflicts and the BIS. However, there is currently no consensus for the direction of the frontal activation. The authors present here a modified version of the T-Maze paradigm, a motivational conflict desktop task, in which behavior is recorded simultaneously to the recording of high-density EEG (HD-EEG). Methods: In this within-subject design, HD-EEG and behavior of 35 healthy participants was recorded. EEG data was collected with a 128 channels sponge-based system. The motivational conflict desktop task consisted of three blocks of repeated trials. Each block was designed to record a slightly different behavioral pattern, to increase the chances of eliciting conflict. This variety of behavioral patterns was however similar enough to allow comparison of the number of trials categorized as ‘overt conflict’ between the blocks. Results: Overt conflict behavior was exhibited in all blocks, but always for under 10% of the trials, in average, in each block. However, changing the order of the paradigms successfully introduced a ‘reset’ of the conflict process, therefore providing more trials for analysis. As for the EEG correlates, the authors expect a different pattern for trials categorized as conflict, compared to the other ones. More specifically, we expect an elevated alpha frequency power in the left frontal electrodes at around 200ms post-cueing, compared to the right one (relative higher right frontal activity), followed by an inversion around 600ms later. Conclusion: With this comprehensive approach of a psychological mechanism, new evidence would be brought to the frontal asymmetry discussion, and its relationship with the BIS. Furthermore, with the present task focusing on a very particular type of motivational approach-avoidance conflict, it would open the door to further variations of the paradigm to introduce different kinds of conflicts involved in AD. Even though its application as a potential biomarker sounds difficult, because of the individual reliability of both the task and peak frequency in the alpha range, we hope to open the discussion for task robustness for neuromodulation and neurofeedback future applications.

Keywords: anxiety, approach-avoidance conflict, behavioral inhibition system, EEG

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Authors: Victoria Brennan, Andrea Mallaburn, Linda Seton

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Science outreach programmes connect school pupils with external agencies to provide activities and experiences that enhance their exposure to science. It can be argued that these programmes not only aim to support teachers with curriculum engagement and promote scientific literacy but also provide pivotal opportunities to spark scientific interest in students. In turn, a further objective of these programmes is to increase awareness of career opportunities within this field. Although outreach work is also often described as a fun and satisfying venture, a plethora of researchers express caution to how successful the processes are to increases engagement post-16 in science. When researching the impact of outreach programmes, it is often student feedback regarding the activities or enrolment numbers to particular science courses post-16, which are generated and analysed. Although this is informative, the longevity of the programme’s impact could be better informed by the teacher’s perceptions; the evidence of which is far more limited in the literature. In addition, there are strong suggestions that teachers can have an indirect impact on a student’s own self-concept. These themes shape the focus and importance of this ongoing research project as it presents the rationale that teachers are under-used resources when it comes to considering the design of science outreach programmes. Therefore, the end result of the research will consist of a presentation of an ‘optimum’ model of outreach. The result of which should be of interest to the wider stakeholders such as universities or private or government organisations who design science outreach programmes in the hope to recruit future scientists. During phase one, questionnaires (n=52) and interviews (n=8) have generated both quantitative and qualitative data. These have been analysed using the Wilcoxon non-parametric test to compare teachers’ perceptions of science outreach interventions and thematic analysis for open-ended questions. Both of these research activities provide an opportunity for a cross-section of teacher opinions of science outreach to be obtained across all educational levels. Therefore, an early draft of the ‘optimum’ model of science outreach delivery was generated using both the wealth of literature and primary data. This final (ongoing) phase aims to refine this model using teacher focus groups to provide constructive feedback about the proposed model. The analysis uses principles of modified Grounded Theory to ensure that focus group data is used to further strengthen the model. Therefore, this research uses a pragmatist approach as it aims to focus on the strengths of the different paradigms encountered to ensure the data collected will provide the most suitable information to create an improved model of sustainable outreach. The results discussed will focus on this ‘optimum’ model and teachers’ perceptions of benefits and drawbacks when it comes to engaging with science outreach work. Although the model is still a ‘work in progress’, it provides both insight into how teachers feel outreach delivery can be a sustainable intervention tool within the classroom and what providers of such programmes should consider when designing science outreach activities.

Keywords: educational partnerships, science education, science outreach, teachers

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Authors: Danny Alexander R. Moreno, Gustavo L. Sartorello, Yuli Andrea P. Bermudez, Richard R. Lobo, Ives Claudio S. Bueno, Augusto H. Gameiro

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Meat production is a significant sector for Brazil's economy; however, the agricultural segment has suffered censure regarding the negative impacts on the environment, which consequently results in climate change. Therefore, it is essential the implementation of nutritional strategies that can improve the environmental performance of livestock. This research aimed to estimate the environmental impact and profitability of the use of yerba mate extract (Ilex paraguariensis) as an additive in the feeding of feedlot lamb. Thirty-six castrated male lambs (average weight of 23.90 ± 3.67 kg and average age of 75 days) were randomly assigned to four experimental diets with different levels of inclusion of yerba mate extract (0, 1, 2, and 4 %) based on dry matter. The animals were confined for fifty-three days and fed with 60:40 corn silage to concentrate ratio. As an indicator of environmental impact, the carbon footprint (CF) was measured as kg of CO₂ equivalent (CO₂-eq) per kg of body weight produced (BWP). The greenhouse gas (GHG) emissions such as methane (CH₄) generated from enteric fermentation, were calculated using the sulfur hexafluoride gas tracer (SF₆) technique; while the CH₄, nitrous oxide (N₂O - emissions generated by feces and urine), and carbon dioxide (CO₂ - emissions generated by concentrate and silage processing) were estimated using the Intergovernmental Panel on Climate Change (IPCC) methodology. To estimate profitability, the gross margin was used, which is the total revenue minus the total cost; the latter is composed of the purchase of animals and food. The boundaries of this study considered only the lamb fattening system. The enteric CH₄ emission from the lamb was the largest source of on-farm GHG emissions (47%-50%), followed by CH₄ and N₂O emissions from manure (10%-20%) and CO₂ emission from the concentrate, silage, and fossil energy (17%-5%). The treatment that generated the least environmental impact was the group with 4% of yerba mate extract (YME), which showed a 3% reduction in total GHG emissions in relation to the control (1462.5 and 1505.5 kg CO₂-eq, respectively). However, the scenario with 1% YME showed an increase in emissions of 7% compared to the control group. In relation to CF, the treatment with 4% YME had the lowest value (4.1 kg CO₂-eq/kg LW) compared with the other groups. Nevertheless, although the 4% YME inclusion scenario showed the lowest CF, the gross margin decreased by 36% compared to the control group (0% YME), due to the cost of YME as a food additive. The results showed that the extract has the potential for use in reducing GHG. However, the cost of implementing this input as a mitigation strategy increased the production cost. Therefore, it is important to develop political strategies that help reduce the acquisition costs of input that contribute to the search for the environmental and economic benefit of the livestock sector.

Keywords: meat production, natural additives, profitability, sheep

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Authors: Despina Giannadaki, Jos Lelieveld, Andrea Pozzer, John Evans

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Outdoor air pollution by fine particles ranks among the top ten global health risk factors that can lead to premature mortality. Epidemiological cohort studies, mainly conducted in United States and Europe, have shown that the long-term exposure to PM2.5 (particles with an aerodynamic diameter less than 2.5μm) is associated with increased mortality from cardiovascular, respiratory diseases and lung cancer. Fine particulates can cause health impacts even at very low concentrations. Previously, no concentration level has been defined below which health damage can be fully prevented. The World Health Organization ambient air quality guidelines suggest an annual mean PM2.5 concentration limit of 10μg/m3. Populations in large parts of the world, especially in East and Southeast Asia, and in the Middle East, are exposed to high levels of fine particulate pollution that by far exceeds the World Health Organization guidelines. The aim of this work is to evaluate the implementation of recent air quality standards for PM2.5 in the EU, the US and other countries worldwide and estimate what measures will be needed to substantially reduce premature mortality. We investigated premature mortality attributed to fine particulate matter (PM2.5) under adults ≥ 30yrs and children < 5yrs, applying a high-resolution global atmospheric chemistry model combined with epidemiological concentration-response functions. The latter are based on the methodology of the Global Burden of Disease for 2010, assuming a ‘safe’ annual mean PM2.5 threshold of 7.3μg/m3. We estimate the global premature mortality by PM2.5 at 3.15 million/year in 2010. China is the leading country with about 1.33 million, followed by India with 575 thousand and Pakistan with 105 thousand. For the European Union (EU) we estimate 173 thousand and the United States (US) 52 thousand in 2010. Based on sensitivity calculations we tested the gains from PM2.5 control by applying the air quality guidelines (AQG) and standards of the World Health Organization (WHO), the EU, the US and other countries. To estimate potential reductions in mortality rates we take into consideration the deaths that cannot be avoided after the implementation of PM2.5 upper limits, due to the contribution of natural sources to total PM2.5 and therefore to mortality (mainly airborne desert dust). The annual mean EU limit of 25μg/m3 would reduce global premature mortality by 18%, while within the EU the effect is negligible, indicating that the standard is largely met and that stricter limits are needed. The new US standard of 12μg/m3 would reduce premature mortality by 46% worldwide, 4% in the US and 20% in the EU. Implementing the AQG by the WHO of 10μg/m3 would reduce global premature mortality by 54%, 76% in China and 59% in India. In the EU and US, the mortality would be reduced by 36% and 14%, respectively. Hence, following the WHO guideline will prevent 1.7 million premature deaths per year. Sensitivity calculations indicate that even small changes at the lower PM2.5 standards can have major impacts on global mortality rates.

Keywords: air quality guidelines, outdoor air pollution, particulate matter, premature mortality

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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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Authors: Andrea L. Arnett, Ann T. Packard, Yolanda I. Garces, Kenneth W. Merrell

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Objective: Pathologic response following neoadjuvant chemoradiation (CRT) has been associated with improved overall survival (OS). Conflicting results have been reported regarding the pathologic predictive value of positron emission tomography (PET) response in patients with stage III lung cancer. The aim of this study was to evaluate the correlation between post-treatment PET response and pathologic response utilizing novel FDG-PET parameters. Methods: This retrospective study included patients with non-metastatic, stage IIIA (N2) NSCLC cancer treated with CRT followed by resection. All patients underwent PET prior to and after neoadjuvant CRT. Univariate analysis was utilized to assess correlations between PET response, nodal clearance, pCR, and near-complete pathologic response (defined as the microscopic residual disease or less). Maximal standard uptake value (SUV), standard uptake ratio (SUR) [normalized independently to the liver (SUR-L) and blood pool (SUR-BP)], metabolic tumor volume (MTV), and total lesion glycolysis (TLG) were measured pre- and post-chemoradiation. Results: A total of 44 patients were included for review. Median age was 61.9 years, and median follow-up was 2.6 years. Histologic subtypes included adenocarcinoma (72.2%) and squamous cell carcinoma (22.7%), and the majority of patients had the T2 disease (59.1%). The rate of pCR and near-complete pathologic response within the primary lesion was 28.9% and 44.4%, respectively. The average reduction in SUVmₐₓ was 9.2 units (range -1.9-32.8), and the majority of patients demonstrated some degree of favorable treatment response. SUR-BP and SUR-L showed a mean reduction of 4.7 units (range -0.1-17.3) and 3.5 units (range –1.7-12.6), respectively. Variation in PET response was not significantly associated with histologic subtype, concurrent chemotherapy type, stage, or radiation dose. No significant correlation was found between pathologic response and absolute change in MTV or TLG. Reduction in SUVmₐₓ and SUR were associated with increased rate of pathologic response (p ≤ 0.02). This correlation was not impacted by normalization of SUR to liver versus mediastinal blood pool. A threshold of > 75% decrease in SUR-L correlated with near-complete response, with a sensitivity of 57.9% and specificity of 85.7%, as well as positive and negative predictive values of 78.6% and 69.2%, respectively (diagnostic odds ratio [DOR]: 5.6, p=0.02). A threshold of >50% decrease in SUR was also significantly associated pathologic response (DOR 12.9, p=0.2), but specificity was substantially lower when utilizing this threshold value. No significant association was found between nodal PET parameters and pathologic nodal clearance. Conclusions: Our results suggest that treatment response to neoadjuvant therapy as assessed on PET imaging can be a predictor of pathologic response when evaluated via SUV and SUR. SUR parameters were associated with higher diagnostic odds ratios, suggesting improved predictive utility compared to SUVmₐₓ. MTV and TLG did not prove to be significant predictors of pathologic response but may warrant further investigation in a larger cohort of patients.

Keywords: lung cancer, positron emission tomography (PET), standard uptake ratio (SUR), standard uptake value (SUV)

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Authors: Tatiane Benvenuti, Fernando Hamerski, Alexandre Giacobbo, Andrea M. Bernardes, Marco A. S. Rodrigues

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Population growth in urban areas has caused damages to the environment, a consequence of the uncontrolled dumping of domestic and industrial wastewater. The capacity of some plants to purify domestic and agricultural wastewater has been demonstrated by several studies. Since natural wetlands have the ability to transform, retain and remove nutrients, constructed wetlands have been used for wastewater treatment. They are widely recognized as an economical, efficient and environmentally acceptable means of treating many different types of wastewater. T. domingensis Pers. species have shown a good performance and low deployment cost to extract, detoxify and sequester pollutants. Constructed Floating Wetlands (CFWs) consist of emergent vegetation established upon a buoyant structure, floating on surface waters. The upper parts of the vegetation grow and remain primarily above the water level, while the roots extend down in the water column, developing an extensive under water-level root system. Thus, the vegetation grows hydroponically, performing direct nutrient uptake from the water column. Biofilm is attached on the roots and rhizomes, and as physical and biochemical processes take place, the system functions as a natural filter. The aim of this study is to diagnose the application of macrophytes in artificial floating in the treatment of domestic sewage in south Brazil. The T. domingensis Pers. plants were placed in a flotation system (polymer structure), in full scale, in a sewage treatment plant. The sewage feed rate was 67.4 m³.d⁻¹ ± 8.0, and the hydraulic retention time was 11.5 d ± 1.3. This CFW treat the sewage generated by 600 inhabitants, which corresponds to 12% of the population served by this municipal treatment plant. During 12 months, samples were collected every two weeks, in order to evaluate parameters as chemical oxygen demand (COD), biochemical oxygen demand in 5 days (BOD5), total Kjeldahl nitrogen (TKN), total phosphorus, total solids, and metals. The average removal of organic matter was around 55% for both COD and BOD5. For nutrients, TKN was reduced in 45.9% what was similar to the total phosphorus removal, while for total solids the reduction was 33%. For metals, aluminum, copper, and cadmium, besides in low concentrations, presented the highest percentage reduction, 82.7, 74.4 and 68.8% respectively. Chromium, iron, and manganese removal achieved values around 40-55%. The use of T. domingensis Pers. in artificial floating for sewage treatment is an effective and innovative alternative in Brazilian sewage treatment systems. The evaluation of additional parameters in the treatment system may give useful information in order to improve the removal efficiency and increase the quality of the water bodies.

Keywords: constructed wetland, floating system, sewage treatment, Typha domingensis Pers.

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Authors: Victor Pena Ribeiro, Caroline Arruda, Jennyfer Andrea Aldana Mejia, Jairo Kenupp Bastos

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Leishmaniasis is a serious health problem around the world. The treatment of infected individuals with pentavalent antimonial drugs is the main therapeutic strategy. However, they present high toxicity and persistence side effects. Therefore, the discovery of new and safe natural-derived therapeutic agents against leishmaniasis is important. Propolis is a resin of viscous consistency produced by Apis mellifera bees from parts of plants. The main types of Brazilian propolis are green, red, yellow and brown. Thus, the aim of this work was to investigate the chemical composition and leishmanicidal properties of a brown propolis (BP). For this purpose, the hydroalcoholic crude extract of BP was obtained and was fractionated by liquid-liquid chromatography. The chemical profile of the extract and its fractions were obtained by HPLC-UV-DAD. The fractions were submitted to preparative HPLC chromatography for isolation of the major compounds of each fraction. They were analyzed by NMR for structural determination. The volatile compounds were obtained by hydrodistillation and identified by GC/MS. Promastigote forms of Leishmania amazonensis were cultivated in M199 medium and then 2×106 parasites.mL-1 were incubated in 96-well microtiter plates with the samples. The BP was dissolved in dimethyl sulfoxide (DMSO) and diluted into the medium, to give final concentrations of 1.56, 3.12, 6.25, 12.5, 25 and 50 µg.mL⁻¹. The plates were incubated at 25ºC for 24 h, and the lysis percentage was determined by using a Neubauer chamber. The bioassays were performed in triplicate, using a medium with 0.5% DMSO as a negative control and amphotericin B as a positive control. The leishimnicidal effect against promastigote forms was also evaluated at the same concentrations. Cytotoxicity experiments also were performed in 96-well plates against normal (CHO-k1) and tumor cell lines (AGP01 and HeLa) using XTT colorimetric method. Phenolic compounds, flavonoids, and terpenoids were identified in brown propolis. The major compounds were identified as follows: p-coumaric acid (24.6%) for a methanolic fraction, Artepelin-C (29.2%) for ethyl acetate fraction and the compounds of hexane fraction are in the process of structural elucidation. The major volatile compounds identified were β-caryophyllene (10.9%), germacrene D (9.7%), nerolidol (10.8%) and spathulenol (8.5%). The propolis did not show cytotoxicity against normal cell lines (CHO) with IC₅₀ > 100 μg.mL⁻¹, whereas the IC₅₀ < 10 μg.mL⁻¹ showed a potential against the AGP01 cell line, propolis did not demonstrate cytotoxicity against HeLa cell lines IC₅₀ > 100 μg.mL⁻¹. In the determination of the leishmanicidal activity, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations of the crude extract caused the lysis of 76% and 45% of promastigote forms of L. amazonensis, respectively. To the amastigote form, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations caused the mortality of 89% and 75% of L. amazonensis, respectively. The IC₅₀ was 2.8 μg.mL⁻¹ to amastigote form and 3.9 μg.mL⁻¹ to promastigote form, showing a promising activity against Leishmania amazonensis.

Keywords: amastigote, brown propolis, cytotoxicity, promastigote

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Authors: Jimena Alarcon, Andrea Llorens, Gabriel Hernandez, Maritza Palma, Lucia Navarrete

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The research has funding from the Government of Chile and is focused on defining the design of rural primary classroom that stimulates creativity. The relevance of the study consists of its capacity to define adequate educational spaces for the implementation of the design-based learning (DBL) methodology. This methodology promotes creativity and teamwork, generating a meaningful learning experience for students, based on the appreciation of their environment and the generation of projects that contribute positively to their communities; also, is an inquiry-based form of learning that is based on the integration of design thinking and the design process into the classroom. The main goal of the study is to define the design characteristics of rural primary school classrooms, associated with the implementation of the DBL methodology. Along with the change in learning strategies, it is necessary to change the educational spaces in which they develop. The hypothesis indicates that a change in the space and equipment of the classrooms based on the emotions of the students will motivate better learning results based on the implementation of a new methodology. In this case, the pedagogical dynamics require an important interaction between the participants, as well as an environment favorable to creativity. Methodologies from Kansei engineering are used to know the emotional variables associated with their definition. The study is done to 50 students between 6 and 10 years old (average age of seven years), 48% of men and 52% women. Virtual three-dimensional scale models and semantic differential tables are used. To define the semantic differential, self-applied surveys were carried out. Each survey consists of eight separate questions in two groups: question A to find desirable emotions; question B related to emotions. Both questions have a maximum of three alternatives to answer. Data were tabulated with IBM SPSS Statistics version 19. Terms referred to emotions are grouped into twenty concepts with a higher presence in surveys. To select the values obtained as part of the implementation of Semantic Differential, a number expected of 'chi-square test (x2)' frequency calculated for classroom space is considered lower limit. All terms over the N expected a cut point, are included to prepare tables for surveys to find a relation between emotion and space. Statistic contrast (Chi-Square) represents significance level ≥ 0, indicator that frequencies appeared are not random. Then, the most representative terms depend on the variable under study: a) definition of textures and color of vertical surfaces is associated with emotions such as tranquility, attention, concentration, creativity; and, b) distribution of the equipment of the rooms, with emotions associated with happiness, distraction, creativity, freedom. The main findings are linked to the generation of classrooms according to diverse DBL team dynamics. Kansei engineering is the appropriate methodology to know the emotions that students want to feel in the classroom space.

Keywords: creativity, design-based learning, education spaces, emotions

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Authors: Pavlina Capkova, Josef Srovnal, Vera Becvarova, Marie Trkova, Zuzana Capkova, Andrea Stefekova, Vaclava Curtisova, Alena Santava, Sarka Vejvalkova, Katerina Adamova, Radek Vodicka

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ASDs are heterogeneous and complex developmental diseases with a significant genetic background. Recurrent CNVs are known to be a frequent cause of ASD. These CNVs can have, however, a variable expressivity which results in a spectrum of phenotypes from asymptomatic to ID/DD/ASD. ASD is associated with ID in ~75% individuals. Various platforms are used to detect pathogenic mutations in the genome of these patients. The performed study is focused on a determination of the frequency of pathogenic mutations in a group of ASD patients and a group of ID/DD patients using various strategies along with a comparison of their detection rate. The possible role of the origin of these mutations in aetiology of ASD was assessed. The study included 35 individuals with ASD and 68 individuals with ID/DD (64 males and 39 females in total), who underwent rigorous genetic, neurological and psychological examinations. Screening for pathogenic mutations involved karyotyping, screening for FMR1 mutations and for metabolic disorders, a targeted MLPA test with probe mixes Telomeres 3 and 5, Microdeletion 1 and 2, Autism 1, MRX and a chromosomal microarray analysis (CMA) (Illumina or Affymetrix). Chromosomal aberrations were revealed in 7 (1 in the ASD group) individuals by karyotyping. FMR1 mutations were discovered in 3 (1 in the ASD group) individuals. The detection rate of pathogenic mutations in ASD patients with a normal karyotype was 15.15% by MLPA and CMA. The frequencies of the pathogenic mutations were 25.0% by MLPA and 35.0% by CMA in ID/DD patients with a normal karyotype. CNVs inherited from asymptomatic parents were more abundant than de novo changes in ASD patients (11.43% vs. 5.71%) in contrast to the ID/DD group where de novo mutations prevailed over inherited ones (26.47% vs. 16.18%). ASD patients shared more frequently their mutations with their fathers than patients from ID/DD group (8.57% vs. 1.47%). Maternally inherited mutations predominated in the ID/DD group in comparison with the ASD group (14.7% vs. 2.86 %). CNVs of an unknown significance were found in 10 patients by CMA and in 3 patients by MLPA. Although the detection rate is the highest when using CMA, recurrent CNVs can be easily detected by MLPA. CMA proved to be more efficient in the ID/DD group where a larger spectrum of rare pathogenic CNVs was revealed. This study determined that maternally inherited highly penetrant mutations and de novo mutations more often resulted in ID/DD without ASD in patients. The paternally inherited mutations could be, however, a source of the greater variability in the genome of the ASD patients and contribute to the polygenic character of the inheritance of ASD. As the number of the subjects in the group is limited, a larger cohort is needed to confirm this conclusion. Inherited CNVs have a role in aetiology of ASD possibly in combination with additional genetic factors - the mutations elsewhere in the genome. The identification of these interactions constitutes a challenge for the future. Supported by MH CZ – DRO (FNOl, 00098892), IGA UP LF_2016_010, TACR TE02000058 and NPU LO1304.

Keywords: autistic spectrum disorders, copy number variant, chromosomal microarray, intellectual disability, karyotyping, MLPA, multiplex ligation-dependent probe amplification

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Authors: Tiffany Riggleman, Andrea Schultheis, Kalyn Jannace, Jerika Taylor, Michelle Nordstrom, Paul F. Pasquina

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Introduction: Posttraumatic Stress (PTS) and Posttraumatic Stress Disorder (PTSD) remain significant problems for military and veteran communities. Symptoms of PTSD often include poor sleep, intrusive thoughts, difficulty concentrating, and trouble with emotional regulation. Unfortunately, despite its high prevalence, service members diagnosed with PTSD often do not seek help, usually because of the perceived stigma surrounding behavioral health care. To help address these challenges, non-pharmacological, therapeutic approaches are being developed to help improve care and enhance compliance. The Service Dog Training Program (SDTP), which involves teaching patients how to train puppies to become mobility service dogs, has been successfully implemented into PTS/PTSD care programs with anecdotal reports of improved outcomes. This study was designed to assess the biopsychosocial effects of SDTP from military beneficiaries with PTS symptoms. Methods: Individuals between the ages of 18 and 65 with PTS symptom were recruited to participate in this prospective study. Each subject completes 4 weeks of baseline testing, followed by 6 weeks of active service dog training (twice per week for one hour sessions) with a professional service dog trainer. Outcome measures included the Posttraumatic Stress Checklist for the DSM-5 (PCL-5), Generalized Anxiety Disorder questionnaire-7 (GAD-7), Patient Health Questionnaire-9 (PHQ-9), social support/interaction, anthropometrics, blood/serum biomarkers, and qualitative interviews. Preliminary analysis of 17 participants examined mean scores on the GAD-7, PCL-5, and PHQ-9, pre- and post-SDTP, and changes were assessed using Wilcoxon Signed-Rank tests. Results: Post-SDTP, there was a statistically significant mean decrease in PCL-5 scores of 13.5 on an 80-point scale (p=0.03) and a significant mean decrease of 2.2 in PHQ-9 scores on a 27 point scale (p=0.04), suggestive of decreased PTSD and depression symptoms. While there was a decrease in mean GAD-7 scores post-SDTP, the difference was not significant (p=0.20). Recurring themes among results from the qualitative interviews include decreased pain, forgetting about stressors, improved sense of calm, increased confidence, improved communication, and establishing a connection with the service dog. Conclusion: Preliminary results of the first 17 participants in this study suggest that individuals who received SDTP had a statistically significant decrease in PTS symptom, as measured by the PCL-5 and PHQ-9. This ongoing study seeks to enroll a total of 156 military beneficiaries with PTS symptoms. Future analyses will include additional psychological outcomes, pain scores, blood/serum biomarkers, and other measures of the social aspects of PTSD, such as relationship satisfaction and sleep hygiene.

Keywords: post-concussive syndrome, posttraumatic stress, service dog, service dog training program, traumatic brain injury

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Authors: Mauro Callejas-Cuervo, Andrea C. Alarcon-Aldana, Andres F. Ruiz-Olaya, Juan C. Alvarez

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Motion capture (MC) is the construction of a precise and accurate digital representation of a real motion. Systems have been used in the last years in a wide range of applications, from films special effects and animation, interactive entertainment, medicine, to high competitive sport where a maximum performance and low injury risk during training and competition is seeking. This paper presents an inertial and magnetic sensor based technological platform, intended for particular amplitude monitoring and telerehabilitation processes considering an efficient cost/technical considerations compromise. Our platform particularities offer high social impact possibilities by making telerehabilitation accessible to large population sectors in marginal socio-economic sector, especially in underdeveloped countries that in opposition to developed countries specialist are scarce, and high technology is not available or inexistent. This platform integrates high-resolution low-cost inertial and magnetic sensors with adequate user interfaces and communication protocols to perform a web or other communication networks available diagnosis service. The amplitude information is generated by sensors then transferred to a computing device with adequate interfaces to make it accessible to inexperienced personnel, providing a high social value. Amplitude measurements of the platform virtual system presented a good fit to its respective reference system. Analyzing the robotic arm results (estimation error RMSE 1=2.12° and estimation error RMSE 2=2.28°), it can be observed that during arm motion in any sense, the estimation error is negligible; in fact, error appears only during sense inversion what can easily be explained by the nature of inertial sensors and its relation to acceleration. Inertial sensors present a time constant delay which acts as a first order filter attenuating signals at large acceleration values as is the case for a change of sense in motion. It can be seen a damped response of platform virtual in other images where error analysis show that at maximum amplitude an underestimation of amplitude is present whereas at minimum amplitude estimations an overestimation of amplitude is observed. This work presents and describes the platform virtual as a motion capture system suitable for telerehabilitation with the cost - quality and precision - accessibility relations optimized. These particular characteristics achieved by efficiently using the state of the art of accessible generic technology in sensors and hardware, and adequate software for capture, transmission analysis and visualization, provides the capacity to offer good telerehabilitation services, reaching large more or less marginal populations where technologies and specialists are not available but accessible with basic communication networks.

Keywords: inertial sensors, joint amplitude measurement, MEMS, telerehabilitation

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Authors: Andrea Pietrelli, Vincenzo Ferrara, Bruno Allard, Francois Buret, Irene Bavasso, Nicola Lovecchio, Francesca Costantini, Firas Khaled

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This paper aims to show some applications of microbial fuel cells (MFCs), an energy harvesting technique, as clean power source to supply low power device for application like wireless sensor network (WSN) for environmental monitoring. Furthermore, MFC can be used directly as biosensor to analyse parameters like pH and temperature or arranged in form of cluster devices in order to use as small power plant. An MFC is a bioreactor that converts energy stored in chemical bonds of organic matter into electrical energy, through a series of reactions catalysed by microorganisms. We have developed a lab-scale terrestrial microbial fuel cell (TMFC), based on soil that acts as source of bacteria and flow of nutrient and a lab-scale waste water microbial fuel cell (WWMFC), where waste water acts as flow of nutrient and bacteria. We performed large series of tests to exploit the capability as biosensor. The pH value has strong influence on the open circuit voltage (OCV) delivered from TMFCs. We analyzed three condition: test A and B were filled with same soil but changing pH from 6 to 6.63, test C was prepared using a different soil with a pH value of 6.3. Experimental results clearly show how with higher pH value a higher OCV was produced; indeed reactors are influenced by different values of pH which increases the voltage in case of a higher pH value until the best pH value of 7 is achieved. The influence of pH on OCV of lab-scales WWMFC was analyzed at pH value of 6.5, 7, 7.2, 7.5 and 8. WWMFCs are influenced from temperature more than TMFCs. We tested the power performance of WWMFCs considering four imposed values of ambient temperature. Results show how power performance increase proportionally with higher temperature values, doubling the output power from 20° to 40°. The best value of power produced from our lab-scale TMFC was equal to 310 μW using peaty soil, at 1KΩ, corresponding to a current of 0.5 mA. A TMFC can supply proper energy to low power devices of a WSN by means of the design of three stages scheme of an energy management system, which adapts voltage level of TMFC to those required by a WSN node, as 3.3V. Using a commercial DC/DC boost converter, that needs an input voltage of 700 mV, the current source of 0.5 mA, charges a capacitor of 6.8 mF until it will have accumulated an amount of charge equal to 700 mV in a time of 10 s. The output stage includes an output switch that close the circuit after a time of 10s + 1.5ms because the converter can boost the voltage from 0.7V to 3.3V in 1.5 ms. Furthermore, we tested in form of clusters connected in series up to 20 WWMFCs, we have obtained a high voltage value as output, around 10V, but low current value. MFC can be considered a suitable clean energy source to be used to supply low power devices as a WSN node or to be used directly as biosensor.

Keywords: energy harvesting, low power electronics, microbial fuel cell, terrestrial microbial fuel cell, waste-water microbial fuel cell, wireless sensor network

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Authors: Zoltán Höhling, Sarah-Lu Oberschelp, Niklas Gilsdorf, Michael Wirsching, Andrea Kuhnert

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A majority of medical students at the University of Freiburg reported stress-related psychosomatic symptoms which are often associated with their studies. International research supports these findings, as medical students worldwide seem to be at special risk for mental health problems. In some countries and institutions, psychologically based interventions that assist medical students in coping with their stressors have been implemented. It turned out that anonymity is an important aspect here. Many students fear a potential damage of reputation when being associated with mental health problems, which may be due to a high level of competitiveness in classes. Therefore, we launched an online-platform where medical students could anonymously seek help and exchange their experiences with fellow students and experts. Medical students of all semesters have access to it through the university’s learning management system (called “ILIAS”). The informative part of the platform consists of exemplary videos showing medical students (actors) who act out scenes that demonstrate the antecedents of stress-related psychosomatic disorders. These videos are linked to different expert comments, describing the exhibited symptoms in an understandable and normalizing way. The (inter-)active part of the platform consists of self-help tools (such as meditation exercises or general tips for stress-coping) and an anonymous interactive forum where students can describe their stress-related problems and seek guidance from experts and/or share their experiences with fellow students. Besides creating an immediate proposal to help affected students, we expect that competitiveness between students might be diminished and bondage improved through mutual support between them. In the initial phase after the platform’s launch, it was accessed by a considerable number of medical students. On a closer look it appeared that platform sections like general information on psychosomatic-symptoms and self-treatment tools were accessed far more often than the online-forum during the first months after the platform launch. Although initial acceptance of the platform was relatively high, students showed a rather passive way of using our platform. While user statistics showed a clear demand for information on stress-related psychosomatic symptoms and its possible remedies, active engagement in the interactive online-forum was rare. We are currently advertising the platform intensively and trying to point out the assured anonymity of the platform and its interactive forum. Our plans, to assure students their anonymity through the use of an e-learning facility and promote active engagement in the online forum, did not (yet) turn out as expected. The reasons behind this may be manifold and based on either e-learning related issues or issues related to students’ individual needs. Students might, for example, question the assured anonymity due to a lack of trust in the technological functioning university’s learning management system. However, one may also conclude that reluctance to discuss stress-related psychosomatic symptoms with peer medical students may not be solely based on anonymity concerns, but could be rooted in more complex issues such as general mistrust between students.

Keywords: e-tutoring, stress-coping, student support, online forum

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Authors: Zubair Ahmed, Andrea Barbieri

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The need to reduce energy consumption has prompted a considerable research effort for developing alternative energy-efficient lighting systems to replace conventional light sources (i.e., incandescent and fluorescent lamps). Organic light emitting device (OLED) technology offers the distinctive possibility to fabricate large area flat devices by vacuum or solution processing. Lanthanide β-diketonates complexes, due to unique photophysical properties of Ln(III) ions, have been explored as emitting layers in OLED displays and in solid-state lighting (SSL) in order to achieve high efficiency and color purity. For such applications, the excellent photoluminescence quantum yield (PLQY) and stability are the two key points that can be achieved simply by selecting the proper organic ligands around the Ln ion in a coordination sphere. Regarding the strategies to enhance the PLQY, the most common is the suppression of the radiationless deactivation pathways due to the presence of high-frequency oscillators (e.g., OH, –CH groups) around the Ln centre. Recently, a different approach to maximize the PLQY of Ln(β-DKs) has been proposed (named 'Escalate Coordination Anisotropy', ECA). It is based on the assumption that coordinating the Ln ion with different ligands will break the centrosymmetry of the molecule leading to less forbidden transitions (loosening the constraints of the Laporte rule). The OLEDs based on such complexes are available, but with low efficiency and stability. In order to get efficient devices, there is a need to develop some new Ln complexes with enhanced PLQYs and stabilities. For this purpose, the Ln complexes, both visible and (NIR) emitting, of variant coordination structures based on the various fluorinated/non-fluorinated β-diketones and O/N-donor neutral ligands were synthesized using a one step in situ method. In this method, the β-diketones, base, LnCl₃.nH₂O and neutral ligands were mixed in a 3:3:1:1 M ratio in ethanol that gave air and moisture stable complexes. Further, they were characterized by means of elemental analysis, NMR spectroscopy and single crystal X-ray diffraction. Thereafter, their photophysical properties were studied to select the best complexes for the fabrication of stable and efficient OLEDs. Finally, the OLEDs were fabricated and investigated using these complexes as emitting layers along with other organic layers like NPB,N,N′-Di(1-naphthyl)-N,N′-diphenyl-(1,1′-biphenyl)-4,4′-diamine (hole-transporting layer), BCP, 2,9-Dimethyl-4,7-diphenyl-1,10-phenanthroline (hole-blocker) and Alq3 (electron-transporting layer). The layers were sequentially deposited under high vacuum environment by thermal evaporation onto ITO glass substrates. Moreover, co-deposition techniques were used to improve charge transport in the devices and to avoid quenching phenomena. The devices show strong electroluminescence at 612, 998, 1064 and 1534 nm corresponding to ⁵D₀ →⁷F₂(Eu), ²F₅/₂ → ²F₇/₂ (Yb), ⁴F₃/₂→ ⁴I₉/₂ (Nd) and ⁴I1₃/₂→ ⁴I1₅/₂ (Er). All the devices fabricated show good efficiency as well as stability.

Keywords: electroluminescence, lanthanides, paramagnetic NMR, photoluminescence

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Authors: Iris Vural Gursel, Andrea Ramirez

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Effective utilization of lignin is an important mean for developing economically profitable biorefineries. Current literature suggests that large amounts of lignin will become available in second generation biorefineries. New conversion technologies will, therefore, be needed to carry lignin transformation well beyond combustion to produce energy, but towards high-value products such as chemicals and transportation fuels. In recent years, significant progress on catalysis has been made to improve transformation of lignin, and new catalytic processes are emerging. In this work, a techno-economic assessment of two of these novel conversion routes and comparison with more established lignin pyrolysis route were made. The aim is to provide insights into the potential performance and potential hotspots in order to guide the experimental research and ease the commercialization by early identifying cost drivers, strengths, and challenges. The lignin conversion routes selected for detailed assessment were: (non-catalytic) lignin pyrolysis as the benchmark, direct hydrodeoxygenation (HDO) of lignin and hydrothermal lignin depolymerisation. Products generated were mixed oxygenated aromatic monomers (MOAMON), light organics, heavy organics, and char. For the technical assessment, a basis design followed by process modelling in Aspen was done using experimental yields. A design capacity of 200 kt/year lignin feed was chosen that is equivalent to a 1 Mt/y scale lignocellulosic biorefinery. The downstream equipment was modelled to achieve the separation of the product streams defined. For determining external utility requirement, heat integration was considered and when possible gasses were combusted to cover heating demand. The models made were used in generating necessary data on material and energy flows. Next, an economic assessment was carried out by estimating operating and capital costs. Return on investment (ROI) and payback period (PBP) were used as indicators. The results of the process modelling indicate that series of separation steps are required. The downstream processing was found especially demanding in the hydrothermal upgrading process due to the presence of significant amount of unconverted lignin (34%) and water. Also, external utility requirements were found to be high. Due to the complex separations, hydrothermal upgrading process showed the highest capital cost (50 M€ more than benchmark). Whereas operating costs were found the highest for the direct HDO process (20 M€/year more than benchmark) due to the use of hydrogen. Because of high yields to valuable heavy organics (32%) and MOAMON (24%), direct HDO process showed the highest ROI (12%) and the shortest PBP (5 years). This process is found feasible with a positive net present value. However, it is very sensitive to the prices used in the calculation. The assessments at this stage are associated with large uncertainties. Nevertheless, they are useful for comparing alternatives and identifying whether a certain process should be given further consideration. Among the three processes investigated here, the direct HDO process was seen to be the most promising.

Keywords: biorefinery, economic assessment, lignin conversion, process design

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Authors: Andrea Miconi

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The research will focus on the results of the Tempus European Project eMEDia dedicated to Cross-Media Journalism. The project is founded by the European Commission as it involves four European partners - IULM University, Tampere University, University of Barcelona, and the Mediterranean network Unimed - and three Tunisian Universities – IPSI La Manouba, Sfax and Sousse – along with the Tunisian Ministry for Higher Education and the National Syndicate of Journalists. The focus on Tunisian condition is basically due to the role played by digital activists in its recent history. The research is dedicated to the relationship between political participation, news-making practices and the spread of social media, as it is affecting Tunisian society. As we know, Tunisia during the Arab Spring had been widely considered as a laboratory for the analysis the use of new technologies for political participation. Nonetheless, the literature about the Arab Spring actually fell short in explaining the genesis of the phenomenon, on the one hand by isolating technologies as a casual factor in the spread of demonstrations, and on the other by analyzing North-African condition through a biased perspective. Nowadays, it is interesting to focus on the consolidation of the information environment three years after the uprisings. And what is relevant, only a close, in-depth analysis of Tunisian society is able to provide an explanation of its history, and namely of the part of digital media in the overall evolution of political system. That is why the research is based on different methodologies: desk stage, interviews, and in-depth analysis of communication practices. Networked journalism is the condition determined by the technological innovation on news-making activities: a condition upon which professional journalist can no longer be considered the only player in the information arena, and a new skill must be developed. Along with democratization, nonetheless, the so-called citizen journalism is also likely to produce some ambiguous effects, such as the lack of professional standards and the spread of information cascades, which may prove to be particularly dangerous in an evolving media market as the Tunisian one. This is why, according to the project, a new profile must be defined, which is able to manage this new condition, and which can be hardly reduced to the parameters of traditional journalistic work. Rather than simply using new devices for news visualization, communication professionals must also be able to dialogue with all new players and to accept the decentralized nature of digital environments. This networked nature of news-making seemed to emerge during the Tunisian revolution, when bloggers, journalists, and activists used to retweet each other. Nonetheless, this intensification of communication exchange was inspired by the political climax of the uprising, while all media, by definition, are also supposed to bring some effects on people’s state of mind, culture and daily life routines. That is why it is worth analyzing the consolidation of these practices in a normal, post-revolutionary situation.

Keywords: cross-media, education, Mediterranean, networked journalism, social media, Tunisia

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Authors: Nicola Milani, Carla Bellissimo, Davide Pogliana, Davide Panzin, Luca Garlaschelli, Giulia Facchinetti, Claudia Casson, Luca Marazzina, Andrea Sartori, Simone Rivaroli, Jeff Konin

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The patient is a 26 year-old male amateur football player from Milan, Italy; (81kg; 185cm; BMI 23.6 kg/m²). He sustained a non-contact anterior cruciate ligament tear to his right knee in June 2021. In September 2021, his right knee ligament was reconstructed using a semitendinosus graft. The injury occurred during a football match on natural grass with typical shoes on a warm day (32 degrees celsius). Playing as a defender he sustained the injury during a change of direction, where the foot was fixated on the grass. He felt pain and was unable to continue playing the match. The surgeon approved his rehabilitation to begin two weeks post-operative. The initial physiotherapist assessment determined performing two training sessions per day within the first three months. In the first three weeks, the pain was 4/10 on Numerical Rating Scale (NRS), no swelling, a range of motion was 0-110°, with difficulty fully extending his knee and minimal quadriceps activation. Crutches were discontinued at four weeks with improved walking. Active exercise, electrostimulator, physical therapy, massages, osteopathy, and passive motion were initiated. At week 6, he completed his first functional movement screen; the score was 16/21 with no pain and no swelling. At week 8, the isokinetic test showed a 23% differential deficit between the two legs in maximum strength (at 90°/s). At week 10, he improved to 15% of injury-induced deficit which suggested he was ready to start running. At week 12, the athlete sustained his first threshold test. At week 16, he performed his first return to sports movement assessment, which revealed a 10% stronger difference between the legs. At week 16, he had his second threshold test. At week 17, his first on-field test revealed a 5% differential deficit between the two legs in the hop test. At week 18, isokinetic test demonstrates that the uninjured leg was 7% stronger than the recovering leg in maximum strength (at 90°/s). At week 20, his second on-field test revealed a 2% difference in hop test; at week 21, his third isokinetic test demonstrated a difference of 5% in maximum strength (at 90°/s). At week 21, he performed his second return to sports movement assessment which revealed a 2% difference between the limbs. Since it was the end of the championship, the team asked him to partake in the playoffs; moreover the player was very motivated to participate in the playoffs also because he was the captain of the team. Together with the player and the team, we decided to let him play even though we were aware of a heightened risk of injury than what is reported in the literature because of two factors: biological recovery times and the results of the tests we performed. In the decision making process about the athlete’s recovery time, it is important to balance the information available from the literature with the desires of the patient to avoid frustration.

Keywords: ACL, football, rehabilitation, return to play

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Authors: Andrea Bikfalvi, Josep Llach, Ferran Lazaro, Bojan Jovanovski

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Due to the continuous need to foster university-business cooperation in both developed and developing countries, some higher education institutions face the challenge of designing, piloting, operating, and consolidating knowledge and technology transfer units. University-business cooperation has different maturity stages worldwide, with some higher education institutions excelling in these practices, but with lots of others that could be qualified as intermediate, or even some situated at the very beginning of their knowledge transfer adventure. These latter face the imminent necessity to formally create the technology transfer unit and to draw its roadmap. The complexity of this operation is due to various aspects that need to align and coordinate, including a major change in mission, vision, structure, priorities, and operations. Qualitative in approach, this study presents 5 case studies, consisting of higher education institutions located in the Western Balkans – 2 in Albania, 2 in Bosnia and Herzegovina, 1 in Montenegro- fully immersed in the entrepreneurial journey of creating their knowledge and technology transfer unit. The empirical evidence is developed in a pan-European project, illustratively called KnowHub (reconnecting universities and enterprises to unleash regional innovation and entrepreneurial activity), which is being implemented in three countries and has resulted in at least 15 pilot cooperation agreements between academia and business. Based on a peer-mentoring approach including more experimented and more mature technology transfer models of European partners located in Spain, Finland, and Austria, a series of initial lessons learned are already available. The findings show that each unit developed its tailor-made approach to engage with internal and external stakeholders, offer value to the academic staff, students, as well as business partners. The latest technology underpinning KnowHub services and institutional commitment are found to be key success factors. Although specific strategies and plans differ, they are based on a general strategy jointly developed and based on common tools and methods of strategic planning and business modelling. The main output consists of providing good practice for designing, piloting, and initial operations of units aiming to fully valorise knowledge and expertise available in academia. Policymakers can also find valuable hints on key aspects considered vital for initial operations. The value of this contribution is its focus on the intersection of three perspectives (service orientation, organisational innovation, business model) since previous research has only relied on a single topic or dual approaches, most frequently in the business context and less frequently in higher education.

Keywords: business model, capacity building, entrepreneurial education, knowledge transfer

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Authors: Cynthia Adlerstein-Grimberg, Andrea Bralic-Echeverría

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An extensive and controversial research debate discusses pedagogical documentation (PD) within early childhood education (ECE) as integral to ECE teachers' professional development. The literature converges in acknowledging that ICT integration in PD can be fundamental for children's and teachers' collaborative learning by making their processes visible and open to reflection. Controversial issues about PD emerge around ICT integration and the use of multimedia applications and platforms, displacing the physical experience involved in this pedagogical practice. Authors argue that online platforms make PD become a passive device to demonstrate accountability and performance. Furthermore, ICT integration would make educators inform children and families of pedagogical processes, positioning them more as consumers instead of involving them in collective thinking and pedagogical decision-making. This article analyses how pedagogical documentation mediated by a multimedia application (MAFApp) allows for the positive strengthening of an ECE pedagogical online community that thinks collectively about learning environments. In doing so, the paper shows how ICT integration supports ECE teachers' collective online thinking, enabling them to move from the controversial version of online PD, where they only act as informers of children's learning and assume a voyeuristic perspective, towards a collective online thinking that builds professional development and supports pedagogical decision-making about learning environments. This article answers How ECE teachers' pedagogical documentation evolves with ICT integration using the MAFApp multimedia application in a national ECE online community. From a posthumanist stance, this paper draws on an 18-month collaborative ethnographic immersion in Chile's unique public ECE online PD community. It develops a unique case study of an online ECE pedagogical community mediated by a multimedia application called MAFApp. This ECE online community includes 32 Chilean public kindergartens, 45 ECE teachers, and 72 assistants, who produced 534 pedagogical documentation. Fieldwork included 35 in-depth interviews, 13 discussion groups, and the constant comparison method for the PD coding. Findings show ICT integration in PD builds collective online thinking that evolves through four moments of growing complexity: 1) teachernalism of built environments, 2) onlookerism of children's anecdotes in learning environments; 3) storytelling of children's place-making, and 4) empowering pedagogies for co-creating learning environments. ICT integration through the MAFApp multimedia application enabled ECE teachers to build collective online thinking, making pedagogies of place visible and engaging children in co-constructing learning environments. This online PD is a continuous professional learning space for ECE teachers, empowering pedagogies of place. In conclusion, ICT integration into PD progressively empowers pedagogies of place in Chilean public ECE. Strengthening collective online thinking using the MAFApp multimedia application sharply contrasts with some recent PD research findings. ICT integration to PD enabled strong collective online thinking. Doing so makes PD operate as a place of professional development, pedagogical reflective encounters, and experimentation while inhabiting their own learning environments with children.

Keywords: early childhood education, ICT integration, multimedia application, online collective thinking, pedagogical documentation, professional development

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Authors: Pawel Kiper, Alfonc Baba, Mahmoud Alhelou, Giorgia Pregnolato, Michela Agostini, Andrea Turolla

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Impairment of cervical spine mobility is often related to pain triggered by musculoskeletal disorders or direct traumatic injuries of the spine. To date, these disorders are assessed with goniometers and inclinometers, which are the most popular devices used in clinical settings. Nevertheless, these technologies usually allow measurement of no more than two-dimensional range of motion (ROM) quotes in static conditions. Conversely, the wide use of motion tracking systems able to measure 3 to 6 degrees of freedom dynamically, while performing standard ROM assessment, are limited due to technical complexities in preparing the setup and high costs. Thus, motion tracking systems are primarily used in research. These systems are an integral part of virtual reality (VR) technologies, which can be used for measuring spine mobility. To our knowledge, the accuracy of VR measure has not yet been studied within virtual environments. Thus, the aim of this study was to test the reliability of a protocol for the assessment of sensorimotor function of the cervical spine in a population of healthy subjects and to compare whether using immersive or non-immersive VR for visualization affects the performance. Both VR assessments consisted of the same five exercises and random sequence determined which of the environments (i.e. immersive or non-immersive) was used as first assessment. Subjects were asked to perform head rotation (right and left), flexion, extension and lateral flexion (right and left side bending). Each movement was executed five times. Moreover, the participants were invited to perform head reaching movements i.e. head movements toward 8 targets placed along a circular perimeter each 45°, visualized one-by-one in random order. Finally, head repositioning movement was obtained by head movement toward the same 8 targets as for reaching and following reposition to the start point. Thus, each participant performed 46 tasks during assessment. Main measures were: ROM of rotation, flexion, extension, lateral flexion and complete kinematics of the cervical spine (i.e. number of completed targets, time of execution (seconds), spatial length (cm), angle distance (°), jerk). Thirty-five healthy participants (i.e. 14 males and 21 females, mean age 28.4±6.47) were recruited for the cervical spine assessment with immersive and non-immersive VR environments. Comparison analysis demonstrated that: head right rotation (p=0.027), extension (p=0.047), flexion (p=0.000), time (p=0.001), spatial length (p=0.004), jerk target (p=0.032), trajectory repositioning (p=0.003), and jerk target repositioning (p=0.007) were significantly better in immersive than non-immersive VR. A regression model showed that assessment in immersive VR was influenced by height, trajectory repositioning (p<0.05), and handedness (p<0.05), whereas in non-immersive VR performance was influenced by height, jerk target (p=0.002), head extension, jerk target repositioning (p=0.002), and by age, head flex/ext, trajectory repositioning, and weight (p=0.040). The results of this study showed higher accuracy of cervical spine assessment when executed in immersive VR. The assessment of ROM and kinematics of the cervical spine can be affected by independent and dependent variables in both immersive and non-immersive VR settings.

Keywords: virtual reality, cervical spine, motion analysis, range of motion, measurement validity

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Authors: Andrea Ghermandi

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Natural treatment systems such as constructed wetlands and waste stabilization ponds are increasingly used to treat water and wastewater from a variety of sources, including stormwater and polluted surface water. The provision of ancillary benefits in the form of cultural ecosystem services makes these systems unique among water and wastewater treatment technologies and greatly contributes to determine their potential role in promoting sustainable water management practices. A quantitative analysis of these benefits, however, has been lacking in the literature. Here, a critical assessment of the recreational and educational benefits in natural treatment systems is provided, which combines observed public use from a survey of managers and operators with estimated public use as obtained using geotagged photos from social media as a proxy for visitation rates. Geographic Information Systems (GIS) are used to characterize the spatial boundaries of 273 natural treatment systems worldwide. Such boundaries are used as input for the Application Program Interfaces (APIs) of two popular photo-sharing websites (Flickr and Panoramio) in order to derive the number of photo-user-days, i.e., the number of yearly visits by individual photo users in each site. The adequateness and predictive power of four univariate calibration models using the crowdsourced data as a proxy for visitation are evaluated. A high correlation is found between photo-user-days and observed annual visitors (Pearson's r = 0.811; p-value < 0.001; N = 62). Standardized Major Axis (SMA) regression is found to outperform Ordinary Least Squares regression and count data models in terms of predictive power insofar as standard verification statistics – such as the root mean square error of prediction (RMSEP), the mean absolute error of prediction (MAEP), the reduction of error (RE), and the coefficient of efficiency (CE) – are concerned. The SMA regression model is used to estimate the intensity of public use in all 273 natural treatment systems. System type, influent water quality, and area are found to statistically affect public use, consistently with a priori expectations. Publicly available information regarding the home location of the sampled visitors is derived from their social media profiles and used to infer the distance they are willing to travel to visit the natural treatment systems in the database. Such information is analyzed using the travel cost method to derive monetary estimates of the recreational benefits of the investigated natural treatment systems. Overall, the findings confirm the opportunities arising from an integrated design and management of natural treatment systems, which combines the objectives of water quality enhancement and provision of cultural ecosystem services through public use in a multi-functional approach and compatibly with the need to protect public health.

Keywords: constructed wetlands, cultural ecosystem services, ecological engineering, waste stabilization ponds

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Authors: Till Fehlauer, Blanche Collin, Bernard Angeletti, Andrea Somogyi, Claire Lallemand, Perrine Chaurand, Cédric Dentant, Clement Levard, Jerome Rose

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Over the last decades, yttrium (Y) has gained importance in high-tech applications. It is an essential part of alloys and compounds used for lasers, displays, or cell phones, for example. Due to its chemical similarities with the lanthanides, Y is often considered a rare earth element (REE). Despite their increased usage, the environmental behavior of REEs remains poorly understood. Especially regarding their interactions with plants, many uncertainties exist. On the one hand, Y is known to have a negative effect on root development and germination, but on the other hand, it appears to promote plant growth at low concentrations. In order to understand these phenomena, a precise knowledge is necessary about how Y is absorbed by the plant and how it is handled once inside the organism. Contradictory studies exist, stating that due to a similar ionic radius, Y and the other REEs might be absorbed through Ca²⁺-channels, while others suspect that Y has a shared pathway with Al³⁺. In this study, laser ablation coupled ICP-MS, and synchrotron-based micro-X-ray fluorescence (µXRF, beamline Nanoscopium, SOLEIL, France) have been used in order to localize Y within the plant tissue and identify associated elements. The plant used in this study is Saxifraga paniculata, a rugged alpine plant that has shown an affinity for Y in previous studies (in prep.). Furthermore, Saxifraga paniculata performs guttation, which means that it possesses phloem sap secreting openings on the leaf surface that serve to regulate root pressure. These so-called hydathodes could provide special insights in elemental transport in plants. The plants have been grown on Y doped soil (500mg/kg DW) for four months. The results showed that Y was mainly concentrated in the roots of Saxifraga paniculata (260 ± 85mg/kg), and only a small amount was translocated to the leaves (10 ± 7.8mg/kg). µXRF analysis indicated that within the root transects, the majority of Y remained in the epidermis and hardly penetrated the stele. Laser ablation coupled ICP-MS confirmed this finding and showed a positive correlation in the roots between Y, Fe, Al, and to a lesser extent Ca. In the stem transect, Y was mainly detected in a hotspot of approximately 40µm in diameter situated in the endodermis area. Within the stem and especially in the hotspot, Y was highly colocalized with Al and Fe. Similar-sized Y hotspots have been detected in/on the leaves. All of them were strongly colocalized with Al and Fe, except for those situated within the hydathodes, which showed no colocalization with any of the measured elements. Accordingly, a relation between Y and Ca during root uptake remains possible, whereas a correlation to Fe and Al appears to be dominant in the aerial parts, suggesting common storage compartments, the formation of complexes, or a shared pathway during translocation.

Keywords: laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), Phytoaccumulation, Rare earth elements, Saxifraga paniculata, Synchrotron-based micro-X-ray fluorescence, Yttrium

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Authors: Hannah Edwards, Femke T. A. Buisman-Pijlman, Adrian Esterman, Craig Phillips, Sandra Orgeig, Andrea Gordon

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Mother-infant bonding is a term which refers to the early emotional connectedness between a mother and her infant. Strong mother-infant bonding promotes higher quality mother and infant interactions including prolonged breastfeeding, secure attachment and increased sensitive parenting and maternal responsiveness. Strengthening of all such interactions leads to improved social behavior, and emotional and cognitive development throughout childhood, adolescence and adulthood. The positive outcomes observed following strong mother-infant bonding emphasize the need to screen new mothers for disrupted mother-infant bonding, and in turn the need for a robust, valid tool to assess mother-infant bonding. A recent scoping review conducted by the research team identified four tools to assess mother-infant bonding, all of which employed self-rating scales. Thus, whilst these tools demonstrated both adequate validity and reliability, they rely on self-reported information from the mother. As such this may reflect a mother’s perception of bonding with their infant, rather than their actual behavior. Therefore, a new tool to assess mother-infant bonding has been developed. The Recorded Interaction Task (RIT) addresses shortcomings of previous tools by employing observational methods to assess bonding. The RIT focusses on the common interaction between mother and infant of changing a nappy, at the target age of 2-6 months, which is visually recorded and then later assessed. Thirteen maternal and seven infant behaviors are scored on the RIT Observation Scoring Sheet, and a final combined score of mother-infant bonding is determined. The aim of the current study was to assess the content validity and inter-rater reliability of the RIT. A panel of six experts with specialized expertise in bonding and infant behavior were consulted. Experts were provided with the RIT Observation Scoring Sheet, a visual recording of a nappy change interaction, and a feedback form. Experts scored the mother and infant interaction on the RIT Observation Scoring Sheet and completed the feedback form which collected their opinions on the validity of each item on the RIT Observation Scoring Sheet and the RIT as a whole. Twelve of the 20 items on the RIT Observation Scoring Sheet were scored ‘Valid’ by all (n=6) or most (n=5) experts. Two items received a ‘Not valid’ score from one expert. The remainder of the items received a mixture of ‘Valid’ and ‘Potentially Valid’ scores. Few changes were made to the RIT Observation Scoring Sheet following expert feedback, including rewording of items for clarity and the exclusion of an item focusing on behavior deemed not relevant for the target infant age. The overall ICC for single rater absolute agreement was 0.48 (95% CI 0.28 – 0.71). Experts (n=6) ratings were less consistent for infant behavior (ICC 0.27 (-0.01 – 0.82)) compared to mother behavior (ICC 0.55 (0.28 – 0.80)). Whilst previous tools employ self-report methods to assess mother-infant bonding, the RIT utilizes observational methods. The current study highlights adequate content validity and moderate inter-rater reliability of the RIT, supporting its use in future research. A convergent validity study comparing the RIT against an existing tool is currently being undertaken to confirm these results.

Keywords: content validity, inter-rater reliability, mother-infant bonding, observational tool, recorded interaction task

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Authors: Juliette Begin, Juliano Colapelle, Andrea Taratanu, Daniel Thirion, Amelie Marsot, Bryan A. Ross

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Chronic obstructive pulmonary disease (COPD), a leading cause of death globally, is characterized by chronic airflow obstruction and acute exacerbations (AECOPDs) that are often triggered by respiratory infections. Pseudomonas aeruginosa (P. aeruginosa), a potentially serious bacterial cause of AECOPDs, is treated with targeted anti-pseudomonal antibiotics. These select few antimicrobials are often used as first-line therapy in patients who are clinically unwell and/or in those suspected of P. aeruginosa-related infection prior to confirmation, potentially contributing to antimicrobial resistance. The present study evaluates prescribing practices in patients with a confirmed sputum history of P. aeruginosa admitted for AECOPD at the McGill University Health Centre (MUHC) and treated with anti-pseudomonal antibiotics. Serum antibiotic concentrations were measured from the same-day peak, trough, and mid-dose blood sampling intervals after reaching steady-state (on or after day 3) and were quantified using ultra-high-performance liquid chromatography (UHPLC). Demographic, clinical, and treatment outcomes were extracted from patient medical charts. Treatment failure was defined by respiratory-related death or mechanical ventilation after ≥3 days of antibiotics; antibiotic therapy extended beyond 2 weeks or a new antibiotic regimen started; or urgent care readmission within 30 days for AECOPD. To date, 9 of 30 planned participants have completed testing: seven received ciprofloxacin, one received meropenem, and one received piperacillin-tazobactam. Due to serum sample batching requirements, the serum ciprofloxacin concentration results for the first 2/8 participants are presented at the time of writing. The first participant had serum levels of 5.45mg/L (T₀), 4.74mg/L (T₅₀), and 4.49mg/L (T₁₀₀), while the second had serum levels of 5mg/L (T₀), 2.6mg/L (T₅₀), and 2.51mg/L (T₁₀₀). Pharmacokinetic parameters Cmax (5.18±0.43mg/L), T₁/₂ (23.56±18.94hours), and AUC (181.9±155.95mg*h/l) were higher than reported monograph values and met target AUC-to-MIC ratio of >125. The patients treated with meropenem and with piperacillin-tazobactam experienced treatment failure. Preliminary results suggest that standard ciprofloxacin dosing in patients experiencing an AECOPD and colonized with P. aeruginosa appears to achieve effective serum concentrations. Final cohort results will inform the pharmacokinetic appropriateness and clinical sufficiency of current AECOPD antimicrobial strategies in P. aeruginosa-colonized patients. This study will guide clinicians in determining the appropriate dosing for AECOPD treatment to achieve therapeutic levels, optimizing outcomes, and minimizing adverse effects. It could also highlight the value of routine antibiotic level monitoring in patients with treatment failure to ensure optimal serum concentrations.

Keywords: acute exacerbation, antimicrobial resistance, chronic obstructive pulmonary disease, pharmacokinetics/pharmacodynamics, Pseudomonas aeruginosa

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Authors: Christina Tran, Lu Khoo, Andrea Waylen

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Introduction: The evolution of the dental consultation from paternalism to partnership has been well documented in developed Western countries. Great emphasis is now placed on the importance of empowering patients to make decisions regarding their care, obtaining informed consent, and maintaining patient privacy and confidentiality. With the majority of communication occurring non-verbally, clinicians often adopt behaviours which suggest an approachable and positive attitude. However, evidence indicates that in Asia, a paternalistic model may be favored in medicine. The power imbalance occurring in doctor-patient relationships worldwide may be exacerbated by various factors in Southeast Asia: the strong hierarchical culture, and the large education gap between doctor and patient. Further insight into this matter can be gained by observing patient-dentist communication in Cambodia. The dentist:population ratio in Cambodia is approximately 1:33,000, with rural areas remaining extremely underserviced. We have carried out an observational study of communication in a voluntary dental clinic in Cambodia with the aim of describing whether the patient-dentist relationship follows a paternalistic or patient-centred model. Method: Over a period of two weeks, two clinicians provided dental care as part of a voluntary program in two Cambodian settings: a temporary, rural clinic and a permanent clinic in Phnom Penh. The clinicians independently recorded their experiences in diaries, making observations on the verbal and non-verbal communication between patients and staff. General observations such as the clinic environment were also made. The diaries were then compared and analyzed using a thematic approach. Results: The overall themes that emerged were regarding the clinic environment, verbal communication, and non-verbal communication. Regarding the clinic environment, the rural clinic was arranged in order to easily direct patients from one dentist to another, with little emphasis on continuous patient care. There was also little consideration for patient privacy: patients were often treated in the presence of many observers, including other waiting patients. However, the permanent clinic was structured to allow greater patient privacy, with continuous patient care occurring throughout the appointment. Regarding verbal communication, there was a strongly paternalistic approach to gaining consent and giving instruction. Patients rarely asked questions regarding their treatment, with dentists doing little to encourage patient involvement. Non-verbal communication between patients and dentists was generally paternalistic, with the dentist often addressing the supine patient from above. Patients often avoided making eye-contact, which may have indicated discomfort or lack of engagement. Both adult and paediatric patients rarely raised verbal concerns regarding pain during treatment, despite displaying non-verbal signs of experiencing pain. Anxious paediatric patients were sometimes managed with physical restraint by their mothers to facilitate treatment. Conclusion: Patient-professional communication in the Cambodian dental setting was observed to be generally paternalistic in nature, although more patient-centred aspects were observed in the established, urban setting. However, it should be noted that these observations are subjective in nature, and that the patients’ actual perceptions of their communication experience were unexplored. Further observations in variety of dental settings in Cambodia are needed before any definitive conclusions can be made.

Keywords: patient-dentist communication, paternalism, patient-centered, non-verbal communication

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Authors: Julia Kapr, Andrea Rossi, Haribaskar Ramachandran, Marius Pollet, Ilka Egger, Selina Dangeleit, Katharina Koch, Jean Krutmann, Ellen Fritsche

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It is widely acknowledged that animal models do not always represent human disease. Especially human brain development is difficult to model in animals due to a variety of structural and functional species-specificities. This causes significant discrepancies between predicted and apparent drug efficacies in clinical trials and their subsequent failure. Emerging alternatives based on 3D in vitro approaches, such as human brain spheres or organoids, may in the future reduce and ultimately replace animal models. Here, we present a human induced pluripotent stem cell (hiPSC)-based 3D neural in a vitro disease model for the Cockayne Syndrome B (CSB). CSB is a rare hereditary disease and is accompanied by severe neurologic defects, such as microcephaly, ataxia and intellectual disability, with currently no treatment options. Therefore, the aim of this study is to investigate the molecular and cellular defects found in neural hiPSC-derived CSB models. Understanding the underlying pathology of CSB enables the development of treatment options. The two CSB models used in this study comprise a patient-derived hiPSC line and its isogenic control as well as a CSB-deficient cell line based on a healthy hiPSC line (IMR90-4) background thereby excluding genetic background-related effects. Neurally induced and differentiated brain sphere cultures were characterized via RNA Sequencing, western blot (WB), immunocytochemistry (ICC) and multielectrode arrays (MEAs). CSB-deficiency leads to an altered gene expression of markers for autophagy, focal adhesion and neural network formation. Cell migration was significantly reduced and electrical activity was significantly increased in the disease cell lines. These data hint that the cellular pathologies is possibly underlying CSB. By induction of autophagy, the migration phenotype could be partially rescued, suggesting a crucial role of disturbed autophagy in defective neural migration of the disease lines. Altered autophagy may also lead to inefficient mitophagy. Accordingly, disease cell lines were shown to have a lower mitochondrial base activity and a higher susceptibility to mitochondrial stress induced by rotenone. Since mitochondria play an important role in neurotransmitter cycling, we suggest that defective mitochondria may lead to altered electrical activity in the disease cell lines. Failure to clear the defective mitochondria by mitophagy and thus missing initiation cues for new mitochondrial production could potentiate this problem. With our data, we aim at establishing a disease adverse outcome pathway (AOP), thereby adding to the in-depth understanding of this multi-faced disorder and subsequently contributing to alternative drug development.

Keywords: autophagy, disease modeling, in vitro, pluripotent stem cells

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Authors: Natalia C. E. S. Nascimento, Mercia L. Oliveira, Fernando R. A. Lima, Leonardo T. C. do Nascimento, Marina B. Silveira, Brigida G. A. Schirmer, Andrea V. Ferreira, Carlos Malamut, Juliana B. da Silva

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Tissue hypoxia is a common characteristic of solid tumors leading to decreased sensitivity to radiotherapy and chemotherapy. In the clinical context, tumor hypoxia assessment employing the positron emission tomography (PET) tracer ¹⁸F-fluoromisonidazole ([¹⁸F]FMISO) is helpful for physicians for planning and therapy adjusting. The aim of this work was to implement the synthesis of 18F-FMISO in a TRACERlab® MXFDG module and also to establish the quality control procedure. [¹⁸F]FMISO was synthesized at Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN/Brazil) using an automated synthesizer (TRACERlab® MXFDG, GE) adapted for the production of [¹⁸F]FMISO. The FMISO chemical standard was purchased from ABX. 18O- enriched water was acquired from Center of Molecular Research. Reagent kits containing eluent solution, acetonitrile, ethanol, 2.0 M HCl solution, buffer solution, water for injections and [¹⁸F]FMISO precursor (dissolved in 2 ml acetonitrile) were purchased from ABX. The [¹⁸F]FMISO samples were purified by Solid Phase Extraction method. The quality requirements of [¹⁸F]FMISO are established in the European Pharmacopeia. According to that reference, quality control of [¹⁸F]FMISO should include appearance, pH, radionuclidic identity and purity, radiochemical identity and purity, chemical purity, residual solvents, bacterial endotoxins, and sterility. The duration of the synthesis process was 53 min, with radiochemical yield of (37.00 ± 0.01) % and the specific activity was more than 70 GBq/µmol. The syntheses were reproducible and showed satisfactory results. In relation to the quality control analysis, the samples were clear and colorless at pH 6.0. The spectrum emission, measured by using a High-Purity Germanium Detector (HPGe), presented a single peak at 511 keV and the half-life, determined by the decay method in an activimeter, was (111.0 ± 0.5) min, indicating no presence of radioactive contaminants, besides the desirable radionuclide (¹⁸F). The samples showed concentration of tetrabutylammonium (TBA) < 50μg/mL, assessed by visual comparison to TBA standard applied in the same thin layer chromatographic plate. Radiochemical purity was determined by high performance liquid chromatography (HPLC) and the results were 100%. Regarding the residual solvents tested, ethanol and acetonitrile presented concentration lower than 10% and 0.04%, respectively. Healthy female mice were injected via lateral tail vein with [¹⁸F]FMISO, microPET imaging studies (15 min) were performed after 2 h post injection (p.i), and the biodistribution was analyzed in five-time points (30, 60, 90, 120 and 180 min) after injection. Subsequently, organs/tissues were assayed for radioactivity with a gamma counter. All parameters of quality control test were in agreement to quality criteria confirming that [¹⁸F]FMISO was suitable for use in non-clinical and clinical trials, following the legal requirements for the production of new radiopharmaceuticals in Brazil.

Keywords: automatic radiosynthesis, hypoxic tumors, pharmacopeia, positron emitters, quality requirements

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