Search results for: cellular stress response

Authors: Deepti Kumari, Upamanyu Hore

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Cities are an example of a human-modified environment with few fragments of urban green spaces, which are widely considered for urban biodiversity. The study aims to address the avifaunal diversity in urban parks based on the park size and their urbanization intensity. Also, understanding the key factors affecting species composition and structure as birds are a good indicator of a healthy ecosystem, and they are sensitive to changes in the environment. A 50 m-long line-transect method is used to survey birds in 39 urban parks in Delhi, India. Habitat variables, including vegetation (percentage of non-native trees, percentage of native trees, top canopy cover, sub-canopy cover, diameter at breast height, ground vegetation cover, shrub height) were measured using the quadrat method along the transect, and disturbance variables (distance from water, distance from road, distance from settlement, park area, visitor rate, and urbanization intensity) were measured using ArcGIS and google earth. We analyzed species data for diversity and richness. We explored the relation of species diversity and richness to habitat variables using the multi-model inference approach. Diversity and richness are found significant in different park sizes and their urbanization intensity. Medium size park supports more diversity, whereas large size park has more richness. However, diversity and richness both declined with increasing urbanization intensity. The result of CCA revealed that species composition in urban parks was positively associated with tree diameter at breast height and distance from the settlement. On the model selection approach, disturbance variables, especially distance from road, urbanization intensity, and visitors are the best predictors for the species richness of birds in urban parks. In comparison, multiple regression analysis between habitat variables and bird diversity suggested that native tree species in the park may explain the diversity pattern of birds in urban parks. Feeding guilds such as insectivores, omnivores, carnivores, granivores, and frugivores showed a significant relation with vegetation variables, while carnivores and scavenger bird species mainly responded with disturbance variables. The study highlights the importance of park size in urban areas and their urbanization intensity. It also indicates that distance from the settlement, distance from the road, urbanization intensity, visitors, diameter at breast height, and native tree species can be important determining factors for bird richness and diversity in urban parks. The study also concludes that the response of feeding guilds to vegetation and disturbance in urban parks varies. Therefore, we recommend that park size and surrounding urban matrix should be considered in order to increase bird diversity and richness in urban areas for designing and planning.

Keywords: diversity, feeding guild, urban park, urbanization intensity

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Authors: Evans V. Mauda, Stefan H. Foord, Thinandavha C. Munyai

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The greatest threat to biodiversity is a loss of habitat through landscape fragmentation and attrition. Land use changes are therefore among the most immediate drivers of species diversity. Urbanization and agriculture are the main drivers of habitat loss and transformation in the Savanna biomes of South Africa. Agricultural expansion and the intensification in particular, take place at the expense of biodiversity and will probably be the primary driver of biodiversity loss in this century. Arthropods show measurable behavioural responses to changing land mosaics at the smallest scale and heterogeneous environments are therefore predicted to support more complex and diverse biological assemblages. Ants are premier soil turners, channelers of energy and dominate insect fauna, while spiders are a mega-diverse group that can regulate other invertebrate populations. This study aims to quantify the response of these two taxa in a rural-urban mosaic of a rapidly developing communal area. The study took place in and around two villages in the north-eastern corner of South Africa. Two replicates for each of the dominant land use categories, viz. urban settlements, dryland cultivation and cattle rangelands, were set out in each of the villages and sampled during the dry and wet seasons for a total of 2 villages × 3 land use categories × 2 seasons = 24 assemblages. Local scale variables measured included vertical and horizontal habitat structure as well as structural and chemical composition of the soil. Ant richness was not affected by land use but local scale variables such as vertical vegetation structure (+) and leaf litter cover (+), although vegetation complexity at lower levels was negatively associated with ant richness. However, ant richness was largely shaped by regional and temporal processes invoking the importance of dispersal and historical processes. Spider species richness was mostly affected by land use and local conditions highlighting their landscape elements. Spider richness did not vary much between villages and across seasons and seems to be less dependent on context or history. There was a considerable amount of variation in spider richness that was not explained and this could be related to factors which were not measured in this study such as temperature and competition. For both ant and spider assemblages the constrained ordination explained 18 % of variation in these taxa. Three environmental variables (leaf litter cover, active carbon and rock cover) were important in explaining ant assemblage structure, while two (sand and leaf litter cover) were important for spider assemblage structure. This study highlights the importance of disturbance (land use activities) and leaf litter with the associated effects on ant and spider assemblages across the study area.

Keywords: ants, assemblages, biosphere, diversity, land use, spiders, urbanization

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Authors: Justin Durham, Chanda Rooney, Robert Mather, Mickie Vanhoy

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The psychological construct of empathy is to understand a person’s cognitive perspective and experience the other person’s emotional state. Deciphering emotional states is conducive for interpreting vicarious pain. Observing others' physical pain activates neural networks related to the actual experience of pain itself. The study addresses empathy as a nonlinear dynamic process of simulation for individuals to understand the mental states of others and experience vicarious pain, exhibiting self-organized criticality. Such criticality follows from a combination of neural networks with an excitatory feedback loop generating bistability to resonate permutated empathy. Cortical networks exhibit diverse patterns of activity, including oscillations, synchrony and waves, however, the temporal dynamics of neurophysiological activities underlying empathic processes remain poorly understood. Mu rhythms are EEG oscillations with dominant frequencies of 8-13 Hz becoming synchronized when the body is relaxed with eyes open and when the sensorimotor system is in idle, thus, mu rhythm synchrony is expected to be highest in baseline conditions. When the sensorimotor system is activated either by performing or simulating action, mu rhythms become suppressed or desynchronize, thus, should be suppressed while observing video clips of painful injuries if previous research on mirror system activation holds. Twelve undergraduates contributed EEG data and survey responses to empathy and psychopathy scales in addition to watching consecutive video clips of sports injuries. Participants watched a blank, black image on a computer monitor before and after observing a video of consecutive sports injuries incidents. Each video condition lasted five-minutes long. A BIOPAC MP150 recorded EEG signals from sensorimotor and thalamocortical regions related to a complex neural network called the ‘pain matrix’. Physical and social pain are activated in this network to resonate vicarious pain responses to processing empathy. Five EEG single electrode locations were applied to regions measuring sensorimotor electrical activity in microvolts (μV) to monitor mu rhythms. EEG signals were sampled at a rate of 200 Hz. Mu rhythm desynchronization was measured via 8-13 Hz at electrode sites (F3 & F4). Data for each participant’s mu rhythms were analyzed via Fast Fourier Transformation (FFT) and multifractal time series analysis.

Keywords: desynchronization, dynamical systems theory, electroencephalography (EEG), empathy, multifractal time series analysis, mu waveform, neurophysiology, pain simulation, social cognition

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Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran

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Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.

Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold

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Authors: Juan Bernal-Martinez, Zoe Quinones-Jurado, Miguel Waldo-Mendoza, Elias Perez

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Introduction and Objective: Ag-TiO2 nanoparticles (NP) have been characterized as effective antibacterial compounds against E. aureous, E. coli, Salmonella and others. Because these nanoparticles have been used in plastic-food containers, there is a concern about the toxicity of Ag-TiO2 NP for higher organisms from protozoan, invertebrates, and mammals. The objective of this study is to evaluate the cytotoxic effect of Ag-TiO2 NP on the survival and swimming behavior of the unicellular organism Paramecium tetraurelia. Material and Methods: Preparation of metallic silver on TiO2 surface was based on chemical reduction route of AgNO3. Aqueous suspension of TiO2 nanoparticles was preparing by adding 5 g of TiO2 to 250 ml of deionized water and followed by sonication for 10 min. The required amount of AgNO3 solutions was added to TiO2 suspension, maintaining heating and stirring. Silver concentration was 0.5, 1.5, 5.0, 25, 35 and 45 % w/w versus TiO2. Paramecium tetraurelia (Carolina Biological, Cat. # 131560) was used as a biological preparation. It was cultured in artificial culture media made as follows: Stigmasterol 5 mg/ml of ethanol, Caseaminoacids 0.3 gr/lt.; KCl 4mM; CaCl2 1mM; MgCl2 100uM and MOPS 1mM, pH 7.3. This media was inoculated with Enterobacter-sp. Paramecium was concentrated after 24 hours of incubation by centrifugation. The pellet of cells was resuspended in 4.1.1 solution prepared as follows (in mM): KCl, 4 mM; CaCl2, 1mM and Trizma, 1mM; pH 7.3. Transmission electron microscopy (TEM) studies were performed to evaluate the appropriate dispersion and topographic distribution AgNPs deposited on TiO2. The experimental solutions were prepared as follows: 50 mg of Polyvinyhlpirolidone were added to 5 ml of 4.1.1. solution. Then, 50 mg of powder 25-Ag-TiO2 was added, mixing for 10 min and sonicated for 60 min. Survival of Paramecium and possible toxic effects after 25-Ag-TiO2 treatment was observed through an inverted microscope. The Paramecium swimming behavior and possible dead cells were recorded for periods of approximately 20-50 seconds by using a digital USB camera adapted to the microscope. Results and Discussion: TEM micrographs demonstrated the topographic distribution of AgNPs deposited on TiO2. 25Ag-TiO2 NP was efficiently dissolved and dispersed in 4.1.1 solution at concentrations from 0.1, 1 and 10 mg/ml. When Paramecium were treated with 25Ag-TiO2 NP at 100 ug/ml, it was observed that cells started swimming backwards. This backward swimming behavior is the typical avoiding reaction of the ciliate in response to a noxious stimulus. After 10 min of incubation, it was observed that Paramecium stopped swimming backwards and exploited. We can argue that this toxic effect of 25Ag-TiO2 NP is probably due to the calcium influx and calcium accumulation during the long-lasting swimming backwards. Conclusions: Here we have demonstrated that 25Ag-TiO2 NP has a specific toxic effect on an organism higher than bacteria such as the protozoan Paremecium. Probably these toxic phenomena could be expected to be observed in a higher organism such as invertebrates and mammals.

Keywords: Ag-TiO2, calcium permeability, cytotoxicity, paramecium

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Authors: Lynn Sheridan, Dennis Alonzo, Hoa Nguyen, Andy Gao, Tracy Durksen

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Disruptions to teaching (e.g., COVID-related) have increased work demands for teachers. There is an opportunity for research to explore evidence-informed steps to support teachers. Collective evidence informs data on teachers’ personal attributes (e.g., self-efficacy beliefs) in the workplace are seen to promote success in teaching and support teacher engagement. Teacher engagement plays a role in students’ learning and teachers’ effectiveness. Engaged teachers are better at overcoming work-related stress, burnout and are more likely to take on active roles. Teachers’ commitment is influenced by a host of personal (e.g., teacher well-being) and environmental factors (e.g., job stresses). The job demands-resources model provided a conceptual basis for examining how teachers’ well-being, and is influenced by job demands and job resources. Job demands potentially evoke strain and exceed the employee’s capability to adapt. Job resources entail what the job offers to individual teachers (e.g., organisational support), helping to reduce job demands. The application of the job demands-resources model involves gathering an evidence-base of and connection to personal attributes (job resources). The study explored the association between constructs (resilience, adaptability, commitment, self/collective efficacy) and a teacher’s engagement with the job. The paper sought to elaborate on the model and determine the associations between key constructs of well-being (resilience, adaptability), commitment, and motivation (self and collective-efficacy beliefs) to teachers’ engagement in teaching. Data collection involved online a multi-dimensional instrument using validated items distributed from 2020-2022. The instrument was designed to identify construct relationships. The participant number was 170. Data Analysis: The reliability coefficients, means, standard deviations, skewness, and kurtosis statistics for the six variables were completed. All scales have good reliability coefficients (.72-.96). A confirmatory factor analysis (CFA) and structural equation model (SEM) were performed to provide measurement support and to obtain latent correlations among factors. The final analysis was performed using structural equation modelling. Several fit indices were used to evaluate the model fit, including chi-square statistics and root mean square error of approximation. The CFA and SEM analysis was performed. The correlations of constructs indicated positive correlations exist, with the highest found between teacher engagement and resilience (r=.80) and the lowest between teacher adaptability and collective teacher efficacy (r=.22). Given the associations; we proceeded with CFA. The CFA yielded adequate fit: CFA fit: X (270, 1019) = 1836.79, p < .001, RMSEA = .04, and CFI = .94, TLI = .93 and SRMR = .04. All values were within the threshold values, indicating a good model fit. Results indicate that increasing teacher self-efficacy beliefs will increase a teacher’s level of engagement; that teacher ‘adaptability and resilience are positively associated with self-efficacy beliefs, as are collective teacher efficacy beliefs. Implications for school leaders and school systems: 1. investing in increasing teachers’ sense of efficacy beliefs to manage work demands; 2. leadership approaches can enhance teachers' adaptability and resilience; and 3. a culture of collective efficacy support. Preparing teachers for now and in the future offers an important reminder to policymakers and school leaders on the importance of supporting teachers’ personal attributes when faced with the challenging demands of the job.

Keywords: collective teacher efficacy, teacher self-efficacy, job demands, teacher engagement

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Authors: Fentie Bishaw Wagayie

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This study was carried out with the objective of studying the response of Farta sheep in feed intake and live weight change when fed on hay supplemented with noug seed cake (NSC), wheat bran (WB), and their mixtures. The digestibility trial of 7 days and 90 days of feeding trial was conducted using 25 intact male Farta sheep with a mean initial live weight of 16.83 ± 0.169 kg. The experimental animals were arranged randomly into five blocks based on the initial live weight, and the five treatments were assigned randomly to each animal in a block. Five dietary treatments used in the experiment comprised of grass hay fed ad libitum (T1), grass hay ad libitum + 300 g DM WB (T2), grass hay ad libitum + 300 g DM (67% WB: 33% NSC mixture) (T3), grass hay ad libitum + 300 g DM (67% NSC: 33% WB) (T4) and 300 g DM/ head/day NSC (T5). Common salt and water were offered ad libitum. The supplements were offered twice daily at 0800 and 1600 hours. The experimental sheep were kept in individual pens. Supplementation of NSC, WB, and their mixtures significantly increased (p < 0.01) the total dry matter (DM) (665.84-788 g/head/day) and (p < 0.001) crude protein (CP) intake. Unsupplemented sheep consumed significantly higher (p < 0.01) grass hay DM (540.5g/head/day) as compared to the supplemented treatments (365.8-488 g/h/d), except T2. Among supplemented sheep, T5 had significantly higher (p < 0.001) CP intake (99.98 g/head/day) than the others (85.52-90.2 g/head/day). Supplementation significantly improved (p < 0.001) the digestibility of CP (66.61-78.9%), but there was no significant effect (p > 0.05) on DM, OM, NDF, and ADF digestibility between supplemented and control treatments. Very low CP digestibility (11.55%) observed in the basal diet (grass hay) used in this study indicated that feeding sole grass hay could not provide nutrients even for the maintenance requirement of growing sheep. Significant final and daily live weight gain (p < 0.001) in the range of 70.11-82.44 g/head/day was observed in supplemented Farta sheep, but unsupplemented sheep lost weight by 9.11g/head/day. Numerically, among the supplemented treatments, sheep supplemented with a higher proportion of NSC in T4 (201 NSC + 99 g WB) gained more weight than the rest, though not statistically significant (p > 0.05). The absence of statistical difference in daily body weight gain between all supplemented sheep indicated that the supplementation of NSC, WB, and their mixtures had similar potential to provide nutrients. Generally, supplementation of NSC, WB, and their mixtures to the basal grass hay diet improved feed conversion ratio, total DM intake, CP intake, and CP digestibility, and it also improved the growth performance with a similar trend for all supplemented Farta sheep over the control group. Therefore, from a biological point of view, to attain the required level of slaughter body weight within a short period of the growing program, sheep producer can use all the supplement types depending upon their local availability, but in the order of priority, T4, T5, T3, and T2, respectively. However, based on partial budget analysis, supplementation of 300 g DM/head /day NSC (T5) could be recommended as profitable for producers with no capital limitation, whereas T4 supplementation (201 g NSC + 99 WB DM/day) is recommended when there is capital scarcity.

Keywords: weight gain, supplement, Farta sheep, hay as basal diet

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Authors: Giulia Fredi, Andrea Dorigato, Luca Fambri, Alessandro Pegoretti

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Thermal energy storage (TES) is the storage of heat for later use, thus filling the gap between energy request and supply. The most widely used materials for TES are the organic solid-liquid phase change materials (PCMs), such as paraffin. These materials store/release a high amount of latent heat thanks to their high specific melting enthalpy, operate in a narrow temperature range and have a tunable working temperature. However, they suffer from a low thermal conductivity and need to be confined to prevent leakage. These two issues can be tackled by confining PCMs with carbon nanotubes (CNTs). TES applications include the buildings industry, solar thermal energy collection and thermal management of electronics. In most cases, TES systems are an additional component to be added to the main structure, but if weight and volume savings are key issues, it would be advantageous to embed the TES functionality directly in the structure. Such multifunctional materials could be employed in the automotive industry, where the diffusion of lightweight structures could complicate the thermal management of the cockpit environment or of other temperature sensitive components. This work aims to produce epoxy/carbon structural laminates containing CNT-stabilized paraffin. CNTs were added to molten paraffin in a fraction of 10 wt%, as this was the minimum amount at which no leakage was detected above the melting temperature (45°C). The paraffin/CNT blend was cryogenically milled to obtain particles with an average size of 50 µm. They were added in various percentages (20, 30 and 40 wt%) to an epoxy/hardener formulation, which was used as a matrix to produce laminates through a wet layup technique, by stacking five plies of a plain carbon fiber fabric. The samples were characterized microstructurally, thermally and mechanically. Differential scanning calorimetry (DSC) tests showed that the paraffin kept its ability to melt and crystallize also in the laminates, and the melting enthalpy was almost proportional to the paraffin weight fraction. These thermal properties were retained after fifty heating/cooling cycles. Laser flash analysis showed that the thermal conductivity through the thickness increased with an increase of the PCM, due to the presence of CNTs. The ability of the developed laminates to contribute to the thermal management was also assessed by monitoring their cooling rates through a thermal camera. Three-point bending tests showed that the flexural modulus was only slightly impaired by the presence of the paraffin/CNT particles, while a more sensible decrease of the stress and strain at break and the interlaminar shear strength was detected. Optical and scanning electron microscope images revealed that these could be attributed to the preferential location of the PCM in the interlaminar region. These results demonstrated the feasibility of multifunctional structural TES composites and highlighted that the PCM size and distribution affect the mechanical properties. In this perspective, this group is working on the encapsulation of paraffin in a sol-gel derived organosilica shell. Submicron spheres have been produced, and the current activity focuses on the optimization of the synthesis parameters to increase the emulsion efficiency.

Keywords: carbon fibers, carbon nanotubes, lightweight materials, multifunctional composites, thermal energy storage

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Authors: Heiko Jessen, Laura Tanus, Slobodan Ruzicic

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Objectives: The efficacy of Stribild®, an integrase strand transfer inhibitor (INSTI) -based STR, has been evaluated in randomized clinical trials and it has demonstrated durable capability in terms of achieving sustained suppression of HIV-1 RNA-levels. However, differences in monitoring frequency, existing selection bias and profile of patients enrolled in the trials, may all result in divergent efficacy of this regimen in routine clinical settings. The aim of this study was to assess the virologic outcomes, safety and tolerability profile of Stribild® in a routine clinical setting. Methods: This was a retrospective monocentric analysis on HIV-1-infected patients, who started with or were switched to Stribild®. Virological failure (VF) was defined as confirmed HIV-RNA>50 copies/ml. The minimum time of follow-up was 24 weeks. The percentage of patients remaining free of therapeutic failure was estimated using the time-to-loss-of-virologic-response (TLOVR) algorithm, by intent-to-treat analysis. Results: We analyzed the data of 197 patients (56 ART-naïve and 141 treatment-experienced patients), who fulfilled the inclusion criteria. Majority (95.9%) of patients were male. The median time of HIV-infection at baseline was 2 months in treatment-naïve and 70 months in treatment-experienced patients. Median time [IQR] under ART in treatment-experienced patients was 37 months. Among the treatment-experienced patients 27.0% had already been treated with a regimen consisting of two NRTIs and one INSTI, whereas 18.4% of them experienced a VF. The median time [IQR] of virological suppression prior to therapy with Stribild® in the treatment-experienced patients was 10 months [0-27]. At the end of follow-up (median 33 months), 87.3% (95% CI, 83.5-91.2) of treatment-naïve and 80.3% (95% CI, 75.8-84.8) of treatment-experienced patients remained free of therapeutic failure. Considering only treatment-experienced patients with baseline VL<50 copies/ml, 83.0% (95% CI, 78.5-87.5) remained free of therapeutic failure. A total of 17 patients stopped treatment with Stribild®, 5.4% (3/56) of them were treatment-naïve and 9.9% (14/141) were treatment-experienced patients. The Stribild® therapy was discontinued in 2 (1.0%) because of VF, loss to follow-up in 4 (2.0%), and drug-drug interactions in 2 (1.0%) patients. Adverse events were in 7 (3.6%) patients the reason to switch from therapy with Stribild® and further 2 (1.0%) patients decided personally to switch. The most frequently observed adverse events were gastrointestinal side effects (20.0%), headache (8%), rash events (7%) and dizziness (6%). In two patients we observed an emergence of novel resistances in integrase-gene. The N155H evolved in one patient and resulted in VF. In another patient S119R evolved either during or shortly upon switch from therapy with Stribild®. In one further patient with VF two novel mutations in the RT-gene were observed when compared to historical genotypic test result (V106I/M and M184V), whereby it is not clear whether they evolved during or already before the switch to Stribild®. Conclusions: Effectiveness of Stribild® for treatment-naïve patients was consistent with data obtained in clinical trials. The safety and tolerability profile as well as resistance development confirmed clinical efficacy of Stribild® in a daily practice setting.

Keywords: ART, HIV, integrase inhibitor, stribild

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Authors: Pedro J. Rolim-Neto, Leslie R. M. Ferraz, Fabiana L. A. Santos, Pablo A. Ferreira, Ricardo T. L. Maia-Jr., Magaly A. M. Lyra, Danilo A F. Fonte, Salvana P. M. Costa, Amanda C. Q. M. Vieira, Larissa A. Rolim

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Initially developed as an antimalarial agent, hydroxychloroquine (HCQ) sulfate is often used as a slow-acting antirheumatic drug in the treatment of disorders of connective tissue. The United States Pharmacopeia (USP) 37 provides a reversed-phase HPLC method for quantification of HCQ. However, this method was not reproducible, producing asymmetric peaks in a long analysis time. The asymmetry of the peak may cause an incorrect calculation of the concentration of the sample. Furthermore, the analysis time is unacceptable, especially regarding the routine of a pharmaceutical industry. The aiming of this study was to develop a fast, easy and efficient method for quantification of HCQ sulfate by High Performance Liquid Chromatography (HPLC) based on the Quality by Design (QbD) methodology. This method was optimized in terms of peak symmetry using the surface area graphic as the Design of Experiments (DoE) and the tailing factor (TF) as an indicator to the Design Space (DS). The reference method used was that described at USP 37 to the quantification of the drug. For the optimized method, was proposed a 33 factorial design, based on the QbD concepts. The DS was created with the TF (in a range between 0.98 and 1.2) in order to demonstrate the ideal analytical conditions. Changes were made in the composition of the USP mobile-phase (USP-MP): USP-MP: Methanol (90:10 v/v, 80:20 v/v and 70:30 v/v), in the flow (0.8, 1.0 and 1.2 mL) and in the oven temperature (30, 35, and 40ºC). The USP method allowed the quantification of drug in a long time (40-50 minutes). In addition, the method uses a high flow rate (1,5 mL.min-1) which increases the consumption of expensive solvents HPLC grade. The main problem observed was the TF value (1,8) that would be accepted if the drug was not a racemic mixture, since the co-elution of the isomers can become an unreliable peak integration. Therefore, the optimization was suggested in order to reduce the analysis time, aiming a better peak resolution and TF. For the optimization method, by the analysis of the surface-response plot it was possible to confirm the ideal setting analytical condition: 45 °C, 0,8 mL.min-1 and 80:20 USP-MP: Methanol. The optimized HPLC method enabled the quantification of HCQ sulfate, with a peak of high resolution, showing a TF value of 1,17. This promotes good co-elution of isomers of the HCQ, ensuring an accurate quantification of the raw material as racemic mixture. This method also proved to be 18 times faster, approximately, compared to the reference method, using a lower flow rate, reducing even more the consumption of the solvents and, consequently, the analysis cost. Thus, an analytical method for the quantification of HCQ sulfate was optimized using QbD methodology. This method proved to be faster and more efficient than the USP method, regarding the retention time and, especially, the peak resolution. The higher resolution in the chromatogram peaks supports the implementation of the method for quantification of the drug as racemic mixture, not requiring the separation of isomers.

Keywords: analytical method, hydroxychloroquine sulfate, quality by design, surface area graphic

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Authors: Yang Yang, Luciana Magalhães Rebelo Alencar, Martha Sahylí Ortega Pijeira, Beatriz da Silva Batista, Alefe Roger Silva França, Erick Rafael Dias Rates, Ruana Cardoso Lima, Sara Gemini-Piperni, Ralph Santos-Oliveira

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Radium-²²³ dichloride ([²²³Rₐ]RₐCl₂) is an alpha particle-emitting radiopharmaceutical currently approved for the treatment of patients with castration-resistant prostate cancer, symptomatic bone metastases, and no known visceral metastatic disease. [²²³Rₐ]RₐCl₂ is bone-seeking calcium mimetic that bonds into the newly formed bone stroma, especially osteoblastic or sclerotic metastases, killing the tumor cells by inducing DNA breaks in a potent and localized manner. Nonetheless, the successful therapy of osteosarcoma as primary bone tumors is still a challenge. Nanomicelles are colloidal nanosystems widely used in drug development to improve blood circulation time, bioavailability, and specificity of therapeutic agents, among other applications. In addition, the enhanced permeability and retention effect of the nanosystems, and the renal excretion of the nanomicelles reported in most cases so far, are very attractive to achieve selective and increased accumulation in tumor site as well as to increase the safety of [²²³Rₐ]RₐCl₂ in the clinical routine. In the present work, [²²³Rₐ]RₐCl₂ nanomicelles were produced, characterized, in vitro evaluated, and compared with pure [²²³Rₐ]RₐCl2 solution using SAOS2 osteosarcoma cells. The [²²³Rₐ]RₐCl₂ nanomicelles were prepared using the amphiphilic copolymer Pluronic F127. The dynamic light scattering analysis of freshly produced [²²³Rₐ]RₐCl₂ nanomicelles demonstrated a mean size of 129.4 nm with a polydispersity index (PDI) of 0.303. After one week stored in the refrigerator, the mean size of the [²²³Rₐ]RₐCl₂ nanomicelles increased to 169.4 with a PDI of 0.381. Atomic force microscopy analysis of [223Rₐ]RₐCl₂ nanomicelles exhibited spherical structures whose heights reach 1 µm, suggesting the filling of 127-Pluronic nanomicelles with [²²³Rₐ]RₐCl₂. The viability assay with [²²³Rₐ]RₐCl₂ nanomicelles displayed a dose-dependent response as it was observed using pure [²²³Rₐ]RₐCl2. However, at the same dose, [²²³Rₐ]RₐCl₂ nanomicelles were 20% higher efficient in killing SAOS2 cells when compared with pure [²²³Rₐ]RₐCl₂. These findings demonstrated the effectiveness of the nanosystem validating the application of nanotechnology in targeted alpha therapy with [²²³Ra]RₐCl₂. In addition, the [²²³Rₐ]RaCl₂nanomicelles may be decorated and incorporated with a great variety of agents and compounds (e.g., monoclonal antibodies, aptamers, peptides) to overcome the limited use of [²²³Ra]RₐCl₂.

Keywords: nanomicelles, osteosarcoma, radium dichloride, targeted alpha therapy

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Authors: Wu Liching

Abstract:

Aim This case aims to discuss the pathogenesis, clinical manifestations and medical treatment of strokes caused by mitochondrial gene mutations. Methods Diagnosis of ischemic stroke caused by mitochondrial gene defect by means of "next-generation sequencing mitochondrial DNA gene variation detection", imaging examination, neurological examination, and medical history; this study took samples from the neurology ward of a medical center in northern Taiwan cases diagnosed with acute cerebral infarction as the research objects. Result This case is a 49-year-old married woman with a rare disease, mitochondrial gene mutation inducing ischemic stroke. She has severe hearing impairment and needs to use hearing aids, and has a history of diabetes. During the patient’s hospitalization, the blood test showed that serum Lactate: 7.72 mmol/L, Lactate (CSF) 5.9 mmol/L. Through the collection of relevant medical history, neurological evaluation showed changes in consciousness and cognition, slow response in language expression, and brain magnetic resonance imaging examination showed subacute bilateral temporal lobe infarction, which was an atypical type of stroke. The lineage DNA gene has m.3243A>G known pathogenic mutation point, and its heteroplasmic level is 24.6%. This pathogenic point is located in MITOMAP and recorded as Mitochondrial Encephalopathy, Lactic Acidosis, and Stroke-like episodes (MELAS) , Leigh Syndrome and other disease-related pathogenic loci, this mutation is located in ClinVar and recorded as Pathogenic (dbSNP: rs199474657), so it is diagnosed as a case of stroke caused by a rare disease mitochondrial gene mutation. After medical treatment, there was no more seizure during hospitalization. After interventional rehabilitation, the patient's limb weakness, poor language function, and cognitive impairment have all improved significantly. Conclusion Mitochondrial disorders can also be associated with abnormalities in psychological, neurological, cerebral cortical function, and autonomic functions, as well as problems with internal medical diseases. Therefore, the differential diagnoses cover a wide range and are not easy to be diagnosed. After neurological evaluation, medical history collection, imaging and rare disease serological examination, atypical ischemic stroke caused by rare mitochondrial gene mutation was diagnosed. We hope that through this case, the diagnosis of rare disease mitochondrial gene variation leading to cerebral infarction will be more familiar to clinical medical staff, and this case report may help to improve the clinical diagnosis and treatment for patients with similar clinical symptoms in the future.

Keywords: acute stroke, MELAS, lactic acidosis, mitochondrial disorders

Procedia PDF Downloads 53

Authors: Waseem Akram, Hafiz Azhar Ali Khan

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Unplanned urbanization together with change in life from simple to more technologically advanced style with flow of rural masses to urban areas has played a vital role in pilling loads of solid wastes in our environment. The cities and towns have expanded beyond boundaries. Even the uncontrolled population expansion has caused the overall environmental burden. Thus, today the indifference remains as one of the biggest trash that has come up due to the non-responsive behavior of the people. Everyday huge amount of solid waste is thrown in the streets, on the roads, parks, and in all those places that are frequently and often visited by the human beings. This behavior based response in many countries of the world has led to serious health concerns and environmental issues. Over 80% of our products that are sold in the market are packed in plastic bags. None of the bags are later recycled but simply become a permanent environment concern that flies, choke lines or are burnt and release toxic gases in the environment or form dumps of heaps. Lack of classification of the daily waste generated from houses and other places lead to worst clogging of the sewerage lines and formation of ponding areas which ultimately favor vector borne disease and sometimes become a cause of transmission of polio virus. Solid waste heaps were checked at different places of the cities. All of the wastes on visual assessments were classified into plastic bags, papers, broken plastic pots, clay pots, steel boxes, wrappers etc. All solid waste dumping sites in the cities and wastes that were thrown outside of the trash containers usually contained wrappers, plastic bags, and unconsumed food products. Insect populations seen in these sites included the house flies, bugs, cockroaches and mosquito larvae breeding in water filled wrappers, containers or plastic bags. The population of the mosquitoes, cockroaches and houseflies were relatively very high in dumping sites close to human population. This population has been associated with cases like dengue, malaria, dysentery, gastro and also to skin allergies during the monsoon and summer season. Thus, dumping of the huge amount of solid wastes in and near the residential areas results into serious environmental concerns, bad smell circulation, and health related issues. In some places, the same waste is burnt to get rid of mosquitoes through smoke which ultimately releases toxic material in the atmosphere. Therefore, a proper environmental strategy is needed to minimize environmental burden and promote concepts of recycled products and thus, reduce the disease burden.

Keywords: solid waste accumulation, disease burden, mosquitoes, vector borne diseases

Procedia PDF Downloads 261

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

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

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

Procedia PDF Downloads 148

Authors: Helen S. Joyner (Melito), Mohammad Anvari

Abstract:

Concentrated emulsions stabilized by proteins are systems of great importance in food, pharmaceutical and cosmetic products. Controlling emulsion rheology is critical for ensuring desired properties during formation, storage, and consumption of emulsion-based products. Studies on concentrated emulsions have focused on rheology of monodispersed systems. However, emulsions used for industrial applications are polydispersed in nature, and this polydispersity is regarded as an important parameter that also governs the rheology of the concentrated emulsions. Therefore, the objective of this study was to characterize rheological (small and large deformation behaviors) and microstructural properties of concentrated emulsions which were not truly monodispersed as usually encountered in food products such as margarines, mayonnaise, creams, spreads, and etc. The concentrated emulsions were prepared at different concentrations of fish gelatin (0.2, 0.4, 0.8% w/v in the whole emulsion system), oil-water ratio 80-20 (w/w), homogenization speed 10000 rpm, and 25oC. Confocal laser scanning microscopy (CLSM) was used to determine the microstructure of the emulsions. To prepare samples for CLSM analysis, FG solutions were stained by Fluorescein isothiocyanate dye. Emulsion viscosity profiles were determined using shear rate sweeps (0.01 to 100 1/s). The linear viscoelastic regions (LVRs) of the emulsions were determined using strain sweeps (0.01 to 100% strain) for each sample. Frequency sweeps were performed in the LVR (0.1% strain) from 0.6 to 100 rad/s. Large amplitude oscillatory shear (LAOS) testing was conducted by collecting raw waveform data at 0.05, 1, 10, and 100% strain at 4 different frequencies (0.5, 1, 10, and 100 rad/s). All measurements were performed in triplicate at 25oC. The CLSM results revealed that increased fish gelatin concentration resulted in more stable oil-in-water emulsions with homogeneous, finely dispersed oil droplets. Furthermore, the protein concentration had a significant effect on emulsion rheological properties. Apparent viscosity and dynamic moduli at small deformations increased with increasing fish gelatin concentration. These results were related to increased inter-droplet network connections caused by increased fish gelatin adsorption at the surface of oil droplets. Nevertheless, all samples showed shear-thinning and weak gel behaviors over shear rate and frequency sweeps, respectively. Lissajous plots, or plots of stress versus strain, and phase lag values were used to determine nonlinear behavior of the emulsions in LAOS testing. Greater distortion in the elliptical shape of the plots followed by higher phase lag values was observed at large strains and frequencies in all samples, indicating increased nonlinear behavior. Shifts from elastic-dominated to viscous dominated behavior were also observed. These shifts were attributed to damage to the sample microstructure (e.g. gel network disruption), which would lead to viscous-type behaviors such as permanent deformation and flow. Unlike the small deformation results, the LAOS behavior of the concentrated emulsions was not dependent on fish gelatin concentration. Systems with different microstructures showed similar nonlinear viscoelastic behaviors. The results of this study provided valuable information that can be used to incorporate concentrated emulsions in emulsion-based food formulations.

Keywords: concentrated emulsion, fish gelatin, microstructure, rheology

Procedia PDF Downloads 258

Authors: Leto Leandra, Guaitini Caterina, Agosti Anna, Del Vecchio Lorenzo, Guarrasi Valeria, Cirlini Martina, Chiancone Benedetta

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To the best of our knowledge, this study represents the first attempt to investigate the in vitro germination response of Rhus coriaria L., and its sprout chemical characterization. Rhus coriaria L., a species belonging to the Anacardiaceae family, is commonly called "sumac" and is cultivated, in different countries of the Mediterranean and the Middle East regions, to produce a spice with a sour taste, obtained from its dried and ground fruits. Moreover, since ancient times, many beneficial properties have been attributed to this plant that has been used, in the traditional medicine of several Asian countries, against various diseases, including liver and intestinal pathologies, ulcers and various inflammatory states. In the recent past, sumac was cultivated in the Southern regions of Italy to treat leather, but its cultivation was abandoned, and currently, sumac plants grow spontaneously in marginal areas. Recently, in Italy, the interest in this species has been growing again, thanks to its numerous properties; thus, it becomes imperative to deepen the knowledge of this plant. In this study, in order to set up an efficient in vitro seed germination protocol, sumac seeds collected from spontaneous plants grown in Sicily, an island in the South of Italy, were, firstly, subjected to different treatments, scarification (mechanical, physical and chemical), cold stratification and imbibition, to break their physical and physiological dormancy, then, treated seeds were in vitro cultured on media with different gibberellic acid (GA3) concentrations. Results showed that, without any treatment, only 5% of in vitro sown seeds germinated, while the germination percentage increased up to 19% after the mechanical scarification. A further significative improvement of germination percentages was recorded after the physical scarification, with (40.5%) or without (36.5%) 8 weeks of cold stratification, especially when seeds were sown on gibberellin enriched cultured media. Vitro-derived sumac sprouts, at different developmental stages, were chemically characterized, in terms of polyphenol and tannin content, as well as for their antioxidant activity, to evaluate this matrix as a potential novel food or as a source of bioactive compounds. Results evidenced how more developed sumac sprouts and, above all, their leaves are a wealthy source of polyphenols (78.4 GAE/g SS) and tannins (21.9 mg GAE/g SS), with marked antioxidant activity. The outcomes of this study will be of support the nursery sector and sumac growers in obtaining a higher number of plants in a shorter time; moreover, the sprout chemical characterization will contribute to the process of considering this matrix as a new source of bioactive compounds and tannins to be used in food and non-food sectors.

Keywords: bioactive compounds, germination pre-treatments, rhus coriaria l., tissue culture

Procedia PDF Downloads 75

Authors: Leto Leandra, Guaitini Caterina, Agosti Anna, Del Vecchio Lorenzo, Guarrasi Valeria, Cirlini Martina, Chiancone Benedetta

Abstract:

To the best of our knowledge, this study represents the first attempt to investigate the in vitro germination response of Rhus coriaria L. and its sprout chemical characterization. Rhus coriaria L., a species belonging to the Anacardiaceae family, is commonly called "sumac” and is cultivated, in different countries of the Mediterranean and the Middle East regions, to produce a spice with a sour taste, obtained from its dried and ground fruits. Moreover, since ancient times, many beneficial properties have been attributed to this plant that has been used, in the traditional medicine of several Asian countries, against various diseases, including liver and intestinal pathologies, ulcers, and various inflammatory states. In the recent past, sumac was cultivated in the Southern regions of Italy to treat leather, but its cultivation was abandoned, and currently, sumac plants grow spontaneously in marginal areas. Recently, in Italy, the interest in this species has been growing again, thanks to its numerous properties; thus, it becomes imperative to deepen the knowledge of this plant. In this study, in order to set up an efficient in vitro seed germination protocol, sumac seeds collected from spontaneous plants grown in Sicily, an island in the South of Italy, were, firstly, subjected to different treatments, scarification (mechanical, physical and chemical), cold stratification and imbibition, to break their physical and physiological dormancy, then, treated seeds were in vitro cultured on media with different gibberellic acid (GA3) concentrations. Results showed that, without any treatment, only 5% of in vitro sown seeds germinated, while the germination percentage increased up to 19% after the mechanical scarification. A further significative improvement of germination percentages was recorded after the physical scarification, with (40.5%) or without (36.5%) 8 weeks of cold stratification, especially when seeds were sown on gibberellin enriched cultured media. Vitro-derived sumac sprouts, at different developmental stages, were chemically characterized, in terms of polyphenol and tannin content, as well as for their antioxidant activity, to evaluate this matrix as a potential novel food or as a source of bioactive compounds. Results evidenced how more developed sumac sprouts and, above all, their leaves are a wealthy source of polyphenols (78.4 GAE/g SS) and tannins (21.9 mg GAE/g SS), with marked antioxidant activity. The outcomes of this study will be of support the nursery sector and sumac growers in obtaining a higher number of plants in a shorter time; moreover, the sprout chemical characterization will contribute to the process of considering this matrix as a new source of bioactive compounds and tannins to be used in food and non-food sectors.

Keywords: bioactive compounds, germination pre-treatments, rhus coriaria l., tissue culture

Procedia PDF Downloads 77

Authors: Samuel N. Forjuoh, Marcia G. Ory, Jaewoong Won, Samuel D. Towne, Suojin Wang, Chanam Lee

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The public health burden of obesity is well established as is the influence of physical activity (PA) on the health and wellness of individuals who are obese. This study examined the influence of selected demographic, health, and socioenvironmental factors on the walking behaviors of middle-aged and older overweight and obese adults. Online and paper surveys were administered to community-dwelling overweight and obese adults aged ≥ 50 years residing in four cities in central Texas and seen by a family physician in the primary care clinic from October 2013 to June 2014. Descriptive statistics were used to characterize participants’ anthropometric and demographic data as well as their health conditions and walking, socioenvironmental, and more broadly defined PA behaviors. Then Pearson chi-square tests were used to assess differences between participants who reported walking the recommended ≥ 150 minutes for any purpose in a typical week as a proxy to meeting the U.S. Centers for Disease Control and Prevention’s PA guidelines and those who did not. Finally, logistic regression was used to predict walking the recommended ≥ 150 minutes for any purpose, controlling for covariates. The analysis was conducted in 2016. Of the total sample (n=253, survey response rate of 6.8%), the majority were non-Hispanic white (81.7%), married (74.5%), male (53.5%), and reported an annual household income of ≥ $50,000 (65.7%). Approximately, half were employed (49.6%), or had at least a college degree (51.8%). Slightly more than 1 in 5 (n=57, 22.5%) reported walking the recommended ≥150 minutes for any purpose in a typical week. The strongest predictors of walking the recommended ≥ 150 minutes for any purpose in a typical week in adjusted analysis were related to education and a high favorable perception of the neighborhood environment. Compared to those with a high school diploma or some college, participants with at least a college degree were five times as likely to walk the recommended ≥ 150 minutes for any purpose (OR=5.55, 95% CI=1.79-17.25). Walking the recommended ≥ 150 minutes for any purpose was significantly associated with participants who disagreed that there were many distracted drivers (e.g., on the cell phone while driving) in their neighborhood (OR=4.08, 95% CI=1.47-11.36) and those who agreed that there are sidewalks or protected walkways (e.g., walking trails) in their neighborhood (OR=3.55, 95% CI=1.10-11.49). Those employed were less likely to walk the recommended ≥ 150 minutes for any purpose compared to those unemployed (OR=0.31, 95% CI=0.11-0.85) as were those who reported some difficulty walking for a quarter of a mile (OR=0.19, 95% CI=0.05-0.77). Other socio-environmental factors such as having care-giver responsibilities for elders, someone to walk with, or a dog in the household as well as Walk Score™ were not significantly associated with walking the recommended ≥ 150 minutes for any purpose in a typical week. Neighborhood perception appears to be an important factor associated with the walking behaviors of middle-aged and older overweight and obese individuals. Enhancing the neighborhood environment (e.g., providing walking trails) may promote walking among these individuals.

Keywords: determinants of walking, obesity, older adults, physical activity

Procedia PDF Downloads 238

Authors: Alexandru-Ion Marinescu

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There exist a plethora of methods in the scientific literature which tackle the well-established task of credit score evaluation. In its most abstract form, a credit scoring algorithm takes as input several credit applicant properties, such as age, marital status, employment status, loan duration, etc. and must output a binary response variable (i.e. “GOOD” or “BAD”) stating whether the client is susceptible to payment return delays. Data imbalance is a common occurrence among financial institution databases, with the majority being classified as “GOOD” clients (clients that respect the loan return calendar) alongside a small percentage of “BAD” clients. But it is the “BAD” clients we are interested in since accurately predicting their behavior is crucial in preventing unwanted loss for loan providers. We add to this whole context the constraint that the algorithm must yield an actual, tractable mathematical formula, which is friendlier towards financial analysts. To this end, we have turned to genetic algorithms and genetic programming, aiming to evolve actual mathematical expressions using specially tailored mutation and crossover operators. As far as data representation is concerned, we employ a very flexible mechanism – LINQ expression trees, readily available in the C# programming language, enabling us to construct executable pieces of code at runtime. As the title implies, they model trees, with intermediate nodes being operators (addition, subtraction, multiplication, division) or mathematical functions (sin, cos, abs, round, etc.) and leaf nodes storing either constants or variables. There is a one-to-one correspondence between the client properties and the formula variables. The mutation and crossover operators work on a flattened version of the tree, obtained via a pre-order traversal. A consequence of our chosen technique is that we can identify and discard client properties which do not take part in the final score evaluation, effectively acting as a dimensionality reduction scheme. We compare ourselves with state of the art approaches, such as support vector machines, Bayesian networks, and extreme learning machines, to name a few. The data sets we benchmark against amount to a total of 8, of which we mention the well-known Australian credit and German credit data sets, and the performance indicators are the following: percentage correctly classified, area under curve, partial Gini index, H-measure, Brier score and Kolmogorov-Smirnov statistic, respectively. Finally, we obtain encouraging results, which, although placing us in the lower half of the hierarchy, drive us to further refine the algorithm.

Keywords: expression trees, financial credit scoring, genetic algorithm, genetic programming, symbolic evolution

Procedia PDF Downloads 101

Authors: Tim Bolle, Franziska Kreimendahl, Thomas Gries, Stefan Jockenhoevel

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The therapeutic treatment of extensive, deep wounds is limited. Autologous split-skin grafts are used as a so-called ‘gold standard’. Most common deficits are the defects at the donor site, the risk of scarring as well as the limited availability and quality of the autologous grafts. The aim of this project is a tissue engineered dermal-epidermal skin replacement to overcome the limitations of the gold standard. A key requirement for the development of such a three-dimensional implant is the formation of a functional capillary-like network inside the implant to ensure a sufficient nutrient and gas supply. Tailored three-dimensional warp knitted spacer fabrics are used to reinforce the mechanically week fibrin gel-based scaffold and further to create a directed in vitro pre-vascularization along the parallel-oriented pile yarns within a co-culture. In this study various three-dimensional warp knitted spacer fabrics were developed in a factorial design to analyze the influence of the machine parameters such as the stitch density and the pattern of the fabric on the scaffold performance and further to determine suitable parameters for a successful fibrin gel-incorporation and a physiological performance of the scaffold. The fabrics were manufactured on a Karl Mayer double-bar raschel machine DR 16 EEC/EAC. A fine machine gauge of E30 was used to ensure a high pile yarn density for sufficient nutrient, gas and waste exchange. In order to ensure a high mechanical stability of the graft, the fabrics were made of biocompatible PVDF yarns. Key parameters such as the pore size, porosity and stress/strain behavior were investigated under standardized, controlled climate conditions. The influence of the input parameters on the mechanical and morphological properties as well as the ability of fibrin gel incorporation into the spacer fabric was analyzed. Subsequently, the pile yarns of the spacer fabrics were colonized with Human Umbilical Vein Endothelial Cells (HUVEC) to analyze the ability of the fabric to further function as a guiding structure for a directed vascularization. The cells were stained with DAPI and investigated using fluorescence microscopy. The analysis revealed that the stitch density and the binding pattern have a strong influence on both the mechanical and morphological properties of the fabric. As expected, the incorporation of the fibrin gel was significantly improved with higher pore sizes and porosities, whereas the mechanical strength decreases. Furthermore, the colonization trials revealed a high cell distribution and density on the pile yarns of the spacer fabrics. For a tailored reinforcing structure, the minimum porosity and pore size needs to be evaluated which still ensures a complete incorporation of the reinforcing structure into the fibrin gel matrix. That will enable a mechanically stable dermal graft with a dense vascular network for a sufficient nutrient and oxygen supply of the cells. The results are promising for subsequent research in the field of reinforcing mechanically weak biological scaffolds and develop functional three-dimensional scaffolds with an oriented pre-vascularization.

Keywords: fibrin-gel, skin replacement, spacer fabric, pre-vascularization

Procedia PDF Downloads 240

Authors: Md Kamrul Hassan

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Adaptation interventions are the common response to manage the vulnerabilities of climate change. The nature of adaptation intervention depends on the degree of vulnerability and the capacity of a society. The coping interventions can take the form of hard adaptation – utilising technologies and capital goods like dykes, embankments, seawalls, and/or soft adaptation – engaging knowledge and information sharing, capacity building, policy and strategy development, and innovation. Hard adaptation is quite capital intensive but provides immediate relief from climate change vulnerabilities. This type of adaptation is not real development, as the investment for the adaptation cannot improve the performance – just maintain the status quo of a social or ecological system, and often lead to maladaptation in the long-term. Maladaptation creates a two-way loss for a society – interventions bring further vulnerability on top of the existing vulnerability and investment for getting rid of the consequence of interventions. Hard adaptation is popular to the vulnerable groups, but it focuses so much on the immediate solution and often ignores the environmental issues and future risks of climate change. On the other hand, soft adaptation is education oriented where vulnerable groups learn how to live with climate change impacts. Soft adaptation interventions build the capacity of vulnerable groups through training, innovation, and support, which might enhance the resilience of a system. In consideration of long-term sustainability, soft adaptation can contribute more to resilience than hard adaptation. Taking a developing society as the study context, this study aims to investigate and understand the effectiveness of the adaptation interventions of the coastal community of Sundarbans mangrove forest in Bangladesh. Applying semi-structured interviews with a range of Sundarbans stakeholders including community residents, tourism demand-supply side stakeholders, and conservation and management agencies (e.g., Government, NGOs and international agencies) and document analysis, this paper reports several key insights regarding climate change adaptation. Firstly, while adaptation interventions may offer a short-term to medium-term solution to climate change vulnerabilities, interventions need to be revised for long-term sustainability. Secondly, soft adaptation offers advantages in terms of resilience in a rapidly changing environment, as it is flexible and dynamic. Thirdly, there is a challenge to communicate to educate vulnerable groups to understand more about the future effects of hard adaptation interventions (and the potential for maladaptation). Fourthly, hard adaptation can be used if the interventions do not degrade the environmental balance and if the investment of interventions does not exceed the economic benefit of the interventions. Overall, the goal of an adaptation intervention should be to enhance the resilience of a social or ecological system so that the system can with stand present vulnerabilities and future risks. In order to be sustainable, adaptation interventions should be designed in such way that those can address vulnerabilities and risks of climate change in a long-term timeframe.

Keywords: adaptation, climate change, maladaptation, resilience, Sundarbans, sustainability, vulnerability

Procedia PDF Downloads 173

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

Abstract:

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

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

Procedia PDF Downloads 89

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

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

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

Procedia PDF Downloads 281

Authors: N. B. Chandrakala

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Domestic violence is one of the most under-reported crimes. Problem with domestic violence is that it is not even considered as abuse in many parts of the world especially certain parts of Asia, Africa and Middle East. It is viewed as “doing the needful”. Domestic violence could be in form of emotional harassment, physical injury or psychological abuse perpetrated by one of the family members to another. It is a worldwide phenomenon mainly targeting women. The acts of violence have terrible negative impact on women. It is also an infringement of women’s rights and can be safely termed as human rights abuse. In cases pertaining to domestic violence, male adults often misuses his authority and power to control another using physical or psychological means. Violence and other forms of abuse are common in domestic violence. Sexual assaults, molestation and battering are common in these cases. Domestic violence is a human rights issue and a serious deterrent to development. Domestic violence could also take place in subtle forms like making the person feel worthless or not giving the victims any personal space or freedom. The problematic aspect is cases of domestic violence are very rarely reported. The majority of the victims are women but children are also made to suffer silently. They are abused and neglected. Their innocent minds are adversely affected with the incidents of domestic violence. According to a report by World Health Organization (WHO), sexual trafficking, female feticide, dowry death, public humiliation and physical torture are some of the most common forms of domestic violence against Indian women. Such acts belie our growth and claim as an economic superpower. It is ironic that we claim to be one of the most rapidly advancing countries in the world and yet we have done hardly anything of note against social hazards like domestic violence. Laws are not that stringent when it comes to reporting acts of domestic violence. Even if the report is filed it turns out to be a long drawn process and not every victim has that much resource to fight till the end. It is also a social taboo to make your family matters public. The big challenge in front now is to enforce it in true sense. Steps that are actually needed; tough laws against domestic violence, speedy execution and change in the mindset of society only then we can expect to have some improvement in such inhuman cases. An effective response to violence must be multi-sectoral; addressing the immediate practical needs of women experiencing abuse; providing long-term follow up and assistance; and focusing on changing those cultural norms, attitudes and legal provisions that promote the acceptance of and even encourage violence against women, and undermine women's enjoyment of their full human rights and freedoms. Hence the responses to the problem must be based on integrated approach. The effectiveness of measures and initiatives will depend on coherence and coordination associated with their design and implementation.

Keywords: domestic violence, human rights, sexual assaults, World Health Organization

Procedia PDF Downloads 529

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

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

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

Procedia PDF Downloads 99

Authors: Harry Einhorn

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This paper aims to explore the integration of Buddhism into educational settings with the goal of fostering the holistic development of children. By incorporating Buddhist principles and practices, educators can create a nurturing environment that cultivates wisdom, compassion, and ethical values in children. The teachings of Buddhism provide valuable insights into mindfulness, compassion, and critical thinking, which can be adapted and applied to educational curricula to enhance children's intellectual, emotional, and moral growth. One of the fundamental aspects of Buddhist philosophy that is particularly relevant to education is the concept of mindfulness. By introducing mindfulness practices, such as meditation and breathing exercises, children can learn to cultivate present-moment awareness, develop emotional resilience, and enhance their ability to concentrate and focus. These skills are essential for effective learning and can contribute to reducing stress and promoting overall well-being in children. Mindfulness practices can also teach children how to manage their emotions and thoughts, promoting self-regulation and creating a positive classroom environment. In addition to mindfulness, Buddhism emphasizes the cultivation of compassion and empathy toward all living beings. Integrating teachings on kindness, empathy, and ethical behavior into the educational framework can help children develop a deep sense of interconnectedness and social responsibility. By engaging children in activities that promote empathy and encourage acts of kindness, such as community service projects and cooperative learning, educators can foster the development of compassionate individuals who are actively engaged in creating a more harmonious and compassionate society. Moreover, Buddhist teachings encourage critical thinking and inquiry, which are crucial skills for intellectual development. By introducing children to fundamental Buddhist concepts such as impermanence, interdependence, and the nature of suffering, educators can engage them in philosophical reflections and broaden their perspectives on life. These teachings promote open-mindedness, curiosity, and a deeper understanding of the interconnectedness of all things. Through the exploration of these concepts, children can develop critical thinking skills and gain insights into the complexities of the world, enabling them to navigate challenges with wisdom and discernment. While integrating Buddhism into education requires sensitivity, cultural awareness, and respect for diverse beliefs and backgrounds, it holds great potential for nurturing the holistic development of children. By incorporating mindfulness practices, fostering compassion and empathy, and promoting critical thinking, Buddhism can contribute to the creation of a more compassionate, inclusive, and harmonious educational environment. This integration can shape well-rounded individuals who are equipped with the necessary skills and qualities to navigate the complexities of the modern world with wisdom, compassion, and resilience. In conclusion, the integration of Buddhism into education offers a valuable framework for cultivating wisdom, compassion, and ethical values in children. By incorporating mindfulness, compassion, and critical thinking into educational practices, educators can create a supportive environment that promotes children's holistic development. By nurturing these qualities, Buddhism can help shape individuals who are not only academically proficient but also morally and ethically responsible, contributing to a more compassionate and harmonious society.

Keywords: Buddhism, education, children, mindfulness

Procedia PDF Downloads 49

Authors: Diane Ghanem, Oscar Covarrubias, Michael Raad, Dawn LaPorte, Babar Shafiq

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Introduction: Standardized exams have long been considered a cornerstone in measuring cognitive competency and academic achievement. Their fixed nature and predetermined scoring methods offer a consistent yardstick for gauging intellectual acumen across diverse demographics. Consequently, the performance of artificial intelligence (AI) in this context presents a rich, yet unexplored terrain for quantifying AI's understanding of complex cognitive tasks and simulating human-like problem-solving skills. Publicly available AI language models such as ChatGPT have demonstrated utility in text generation and even problem-solving when provided with clear instructions. Amidst this transformative shift, the aim of this study is to assess ChatGPT’s performance on the orthopaedic surgery in-training examination (OITE). Methods: All 213 OITE 2021 web-based questions were retrieved from the AAOS-ResStudy website. Two independent reviewers copied and pasted the questions and response options into ChatGPT Plus (version 4.0) and recorded the generated answers. All media-containing questions were flagged and carefully examined. Twelve OITE media-containing questions that relied purely on images (clinical pictures, radiographs, MRIs, CT scans) and could not be rationalized from the clinical presentation were excluded. Cohen’s Kappa coefficient was used to examine the agreement of ChatGPT-generated responses between reviewers. Descriptive statistics were used to summarize the performance (% correct) of ChatGPT Plus. The 2021 norm table was used to compare ChatGPT Plus’ performance on the OITE to national orthopaedic surgery residents in that same year. Results: A total of 201 were evaluated by ChatGPT Plus. Excellent agreement was observed between raters for the 201 ChatGPT-generated responses, with a Cohen’s Kappa coefficient of 0.947. 45.8% (92/201) were media-containing questions. ChatGPT had an average overall score of 61.2% (123/201). Its score was 64.2% (70/109) on non-media questions. When compared to the performance of all national orthopaedic surgery residents in 2021, ChatGPT Plus performed at the level of an average PGY3. Discussion: ChatGPT Plus is able to pass the OITE with a satisfactory overall score of 61.2%, ranking at the level of third-year orthopaedic surgery residents. More importantly, it provided logical reasoning and justifications that may help residents grasp evidence-based information and improve their understanding of OITE cases and general orthopaedic principles. With further improvements, AI language models, such as ChatGPT, may become valuable interactive learning tools in resident education, although further studies are still needed to examine their efficacy and impact on long-term learning and OITE/ABOS performance.

Keywords: artificial intelligence, ChatGPT, orthopaedic in-training examination, OITE, orthopedic surgery, standardized testing

Procedia PDF Downloads 67

Authors: Noga Bregman

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Accurate and efficient earthquake detection and phase picking are crucial for seismic hazard assessment and emergency response. This study introduces EQMamba, a deep-learning method that combines the strengths of the Earthquake Transformer and the Mamba model for simultaneous earthquake detection and phase picking. EQMamba leverages the computational efficiency of Mamba layers to process longer seismic sequences while maintaining a manageable model size. The proposed architecture integrates convolutional neural networks (CNNs), bidirectional long short-term memory (BiLSTM) networks, and Mamba blocks. The model employs an encoder composed of convolutional layers and max pooling operations, followed by residual CNN blocks for feature extraction. Mamba blocks are applied to the outputs of BiLSTM blocks, efficiently capturing long-range dependencies in seismic data. Separate decoders are used for earthquake detection, P-wave picking, and S-wave picking. We trained and evaluated EQMamba using a subset of the STEAD dataset, a comprehensive collection of labeled seismic waveforms. The model was trained using a weighted combination of binary cross-entropy loss functions for each task, with the Adam optimizer and a scheduled learning rate. Data augmentation techniques were employed to enhance the model's robustness. Performance comparisons were conducted between EQMamba and the EQTransformer over 20 epochs on this modest-sized STEAD subset. Results demonstrate that EQMamba achieves superior performance, with higher F1 scores and faster convergence compared to EQTransformer. EQMamba reached F1 scores of 0.8 by epoch 5 and maintained higher scores throughout training. The model also exhibited more stable validation performance, indicating good generalization capabilities. While both models showed lower accuracy in phase-picking tasks compared to detection, EQMamba's overall performance suggests significant potential for improving seismic data analysis. The rapid convergence and superior F1 scores of EQMamba, even on a modest-sized dataset, indicate promising scalability for larger datasets. This study contributes to the field of earthquake engineering by presenting a computationally efficient and accurate method for simultaneous earthquake detection and phase picking. Future work will focus on incorporating Mamba layers into the P and S pickers and further optimizing the architecture for seismic data specifics. The EQMamba method holds the potential for enhancing real-time earthquake monitoring systems and improving our understanding of seismic events.

Keywords: earthquake, detection, phase picking, s waves, p waves, transformer, deep learning, seismic waves

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

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

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

Procedia PDF Downloads 288

Authors: Hawraa Zbeeb, Hala Khalifeh, Mohamad Khalil, Francesca Storace, Francesca Baldini, Giulio Lupidi, Laura Vergani

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Sarcopoterium spinosum, a widely distributed spiny shrub belonging to the Rosaceae family, is rich in essential and beneficial constituents. In fact, S. spinosum fruits and roots are traditionally used as herbal medicine in the eastern Mediterranean landscape, and this shrub is mentioned as a medicinal plant in a large number of ethnobotanical surveys. Aqueous root extracts from S. spinosum are used by traditional medicinal practitioners for weight loss treatment of diabetes and pain. Moreover, the anti-diabetic activity of S. spinosum root extract has been reported in different studies, but the beneficial effects of aerial parts, especially fruits, have not been elucidated yet. The aim of the present study was to investigate the in vitro antioxidant and lipid-lowering properties of an ethanolic extract from S. spinosum fruits using both hepatic (FaO) and endothelial (HECV) cells in an attempt to evaluate its possible employment as a nutraceutical supplement. First of all, in vitro spectrophotometric assays were employed to characterize the extract. The total phenol content (TPC) was evaluated by Folin–Ciocalteu spectrophotometric method and the radical scavenging activity was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. After that, the beneficial effects of the extract were tested on cells. FaO cells treated for 3 hours with 0.75 mM oleate/palmitate mix (1:2 molar ratio) mimic in vitro a moderate hepato-steatosis. HECV cells exposed for 1 hour to 100 µM H₂O₂ mimic an oxidative insult leading to oxidative stress conditions. After the metabolic and oxidative insult, both cell lines were treated with increasing concentrations of the S. spinosum extract (1, 10, 25 µg/mL) for 24 hours. The results showed the S. spinosum ethanolic extract is rather rich in phenols (TPC of 18.6 mgGAE/g dry extracts). Moreover, the extract showed a good scavenging ability in vitro (IC₅₀ 15.9 µg/ml and 10.9 µg/ml measured by DPPH and ABTS assays, respectively). When the extract was tested on cells, the results showed that it could ameliorate some markers of cell dysfunction. The three concentrations of the extract led to a significant decrease in the intracellular triglyceride (TG) content in steatotic FaO cells measured by spectrophotometric assay. On the other hand, HECV cells treated with increasing concentrations of the extract did not result in a significant decrease in both lipid peroxidation measured by the Thiobarbituric Acid Reactive Substances (TBARS) assay, and in reactive oxygen species (ROS) production measured by fluorometric analysis after DCF staining. Interestingly, the ethanolic extract was able to accelerate the wound repair of confluent HECV cells with respect to H₂O₂-insulted cells as measured by T-scratch assay. Taken together, these results seem to indicate that the ethanol extract from S. spinosum fruits is rich in phenol compounds and plays considerable lipid-lowering activity in vitro on steatotic hepatocytes and accelerates wound healing repair on endothelial cells. In light of that, the ethanolic extract from S. spinosum fruits could be a potential candidate for nutraceutical applications.

Keywords: antioxidant activity, ethanolic extract, lipid-lowering activity, phenolic compounds, Sarcopoterium spinosum fruits

Procedia PDF Downloads 159