Search results for: Elisa Gonzalez

Authors: Carolina Cuéllar, Paz González

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Research has shown that school leadership has a significant indirect effect on students’ achievements. In Chile, evidence has also revealed that this impact is stronger in vulnerable schools. With the aim of strengthening school leadership, public policy has taken up the challenge of enhancing capabilities of novice principals through the implementation of induction programs, which include a mentoring component, entrusting the task of delivering these programs to universities. The importance of using mentoring or coaching models in the preparation of novice school leaders has been emphasized in the international literature. Thus, it can be affirmed that building leadership capacity through partnership is crucial to facilitate cognitive and affective support required in the initial phase of the principal career, gain role clarification and socialization in context, stimulate reflective leadership practice, among others. In Chile, mentoring is a recent phenomenon in the field of school leadership and it is even more new in the preparation of new principals who work in public schools. This study, funded by the Chilean Ministry of Education, sought to explore the challenges and lessons arising from the design and implementation of mentoring processes which are part of the induction programs, according to the perception of the different actors involved: ministerial agents, university coordinators, mentors and novice principals. The investigation used a qualitative design, based on a study of three cases (three induction programs). The sources of information were 46 semi-structured interviews, applied in two moments (at the beginning and end of mentoring). Content analysis technique was employed. Data focused on the uniqueness of each case and the commonalities within the cases. Five main challenges and lessons emerged in the design and implementation of mentoring within the induction programs for new principals from Chilean public schools. They comprised the need of (i) developing a shared conceptual framework on mentoring among the institutions and actors involved, which helps align the expectations for the mentoring component within the induction programs, along with assisting in establishing a theory of action of mentoring that is relevant to the public school context; (ii) recognizing trough actions and decisions at different levels that the role of a mentor differs from the role of a principal, which challenge the idea that an effective principal will always be an effective mentor; iii) improving mentors’ selection and preparation processes trough the definition of common guiding criteria to ensure that a mentor takes responsibility for developing critical judgment of novice principals, which implies not limiting the mentor’s actions to assist in the compliance of prescriptive practices and standards; (iv) generating common evaluative models with goals, instruments and indicators consistent with the characteristics of mentoring processes, which helps to assess expected results and impact; and (v) including the design of a mentoring structure as an outcome of the induction programs, which helps sustain mentoring within schools as a collective professional development practice. Results showcased interwoven elements that entail continuous negotiations at different levels. Taking action will contribute to policy efforts aimed at professionalizing the leadership role in public schools.

Keywords: induction programs, mentoring, novice principals, school leadership preparation

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Authors: Marina Sofia, Alexios Giannakopoulos, Antonia Touloudi, Dimitris C Chatzopoulos, Zoi Athanasakopoulou, Vassiliki Spyrou, Charalambos Billinis

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Fertility of dairy cattle can be affected by heat stress when the ambient temperature increases above 30°C and the relative humidity ranges from 35% to 50%. The present study was conducted on dairy cattle farms during summer months in Greece and aimed to identify the serological profile against pathogens that could affect fertility and to associate the positive serological results at herd level with temperature factors. A total of 323 serum samples were collected from clinically healthy dairy cows of 8 herds, located in Central and Northern Greece. ELISA tests were performed to detect antibodies against selected pathogens that affect fertility, namely Chlamydophila abortus, Coxiella burnetii, Neospora caninum, Toxoplasma gondii and Infectious Bovine Rhinotracheitis Virus (IBRV). Eleven climatic variables were derived from the WorldClim version 1.4. and ArcGIS V.10.1 software was used for analysis of the spatial information. Five different MaxEnt models were applied to associate the temperature variables with the locations of seropositive Chl. abortus, C. burnetii, N. caninum, T. gondii and IBRV herds (one for each pathogen). The logistic outputs were used for the interpretation of the results. ROC analyses were performed to evaluate the goodness of fit of the models’ predictions. Jackknife tests were used to identify the variables with a substantial contribution to each model. The seropositivity rates of pathogens varied among the 8 herds (0.85-4.76% for Chl. abortus, 4.76-62.71% for N. caninum, 3.8-43.47% for C. burnetii, 4.76-39.28% for T. gondii and 47.83-78.57% for IBRV). The variables of annual temperature range, mean diurnal range and maximum temperature of the warmest month gave a contribution to all five models. The regularized training gains, the training AUCs and the unregularized training gains were estimated. The mean diurnal range gave the highest gain when used in isolation and decreased the gain the most when it was omitted in the two models for seropositive Chl.abortus and IBRV herds. The annual temperature range increased the gain when used alone and decreased the gain the most when it was omitted in the models for seropositive C. burnetii, N. caninum and T. gondii herds. In conclusion, antibodies against Chl. abortus, C. burnetii, N. caninum, T. gondii and IBRV were detected in most herds suggesting circulation of pathogens that could cause infertility. The results of the spatial analyses demonstrated that the annual temperature range, mean diurnal range and maximum temperature of the warmest month could affect positively the possible pathogens’ presence. Acknowledgment: This research has been co‐financed by the European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call RESEARCH–CREATE–INNOVATE (project code: T1EDK-01078).

Keywords: dairy cows, seropositivity, spatial analysis, temperature factors

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Authors: Elisa Boatti, Ulisse Stefanelli, Alessandro Reali, Ferdinando Auricchio

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Thanks to their unusual properties, shape memory alloys (SMAs) are good candidates for advanced applications in a wide range of engineering fields, such as automotive, robotics, civil, biomedical, aerospace. In the last decades, the ever-growing interest for such materials has boosted several research studies aimed at modeling their complex nonlinear behavior in an effective and robust way. Since the constitutive response of SMAs is strongly thermomechanically coupled, the investigation of the non-isothermal evolution of the material must be taken into consideration. The present study considers an existing three-dimensional phenomenological model for SMAs, able to reproduce the main SMA properties while maintaining a simple user-friendly structure, and proposes a variational reformulation of the full non-isothermal version of the model. While the considered model has been thoroughly assessed in an isothermal setting, the proposed formulation allows to take into account the full nonisothermal problem. In particular, the reformulation is inspired to the GENERIC (General Equations for Non-Equilibrium Reversible-Irreversible Coupling) formalism, and is based on a generalized gradient flow of the total entropy, related to thermal and mechanical variables. Such phrasing of the model is new and allows for a discussion of the model from both a theoretical and a numerical point of view. Moreover, it directly implies the dissipativity of the flow. A semi-implicit time-discrete scheme is also presented for the fully coupled thermomechanical system, and is proven unconditionally stable and convergent. The correspondent algorithm is then implemented, under a space-homogeneous temperature field assumption, and tested under different conditions. The core of the algorithm is composed of a mechanical subproblem and a thermal subproblem. The iterative scheme is solved by a generalized Newton method. Numerous uniaxial and biaxial tests are reported to assess the performance of the model and algorithm, including variable imposed strain, strain rate, heat exchange properties, and external temperature. In particular, the heat exchange with the environment is the only source of rate-dependency in the model. The reported curves clearly display the interdependence between phase transformation strain and material temperature. The full thermomechanical coupling allows to reproduce the exothermic and endothermic effects during respectively forward and backward phase transformation. The numerical tests have thus demonstrated that the model can appropriately reproduce the coupled SMA behavior in different loading conditions and rates. Moreover, the algorithm has proved effective and robust. Further developments are being considered, such as the extension of the formulation to the finite-strain setting and the study of the boundary value problem.

Keywords: generalized gradient flow, GENERIC formalism, shape memory alloys, thermomechanical coupling

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Authors: R. P. Hewawasam, M. H. A. D. de Silva, M. A. G. Iresha

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In recent years childhood obesity has increased to pan-epidemic proportions along with a concomitant increase in obesity-associated morbidity. Birth weight is an important determinant of later adult health, with neonates at both ends of the birth weight spectrum at risk of future health complications. Consequently, infants who are born large for gestational age (LGA) are more likely to be obese in childhood and adolescence and are at risk of cardiovascular and metabolic complications later in life. Adipose tissue plays a role in linking events in fetal growth to the subsequent development of adult diseases. In addition to its role as a storage depot for fat, adipose tissue produces and secrets a number of hormones of importance in modulating metabolism and energy homeostasis. Cord blood leptin level has been positively correlated with fetal adiposity at birth. It is established that Asians have lower skeletal muscle mass, low bone mineral content and excess body fat for a given body mass index indicating a genetic predisposition in the occurrence of obesity. To our knowledge, studies have never been conducted in Sri Lanka to determine the relationship between adipocytokine profile in cord blood and anthropometric parameters in newborns. Thus, the objective of this study is to establish the above relationship for the Sri Lankan population to implement awareness programs to minimize childhood obesity in the future. Umbilical cord blood was collected from 90 newborns (Male 40, Female 50; gestational age 35-42 weeks) after double clamping the umbilical cord before separation of the placenta and the concentration of leptin was measured by ELISA technique. Anthropometric parameters of the newborn such as birth weight, length, ponderal index, occipital frontal, chest, hip and calf circumferences were measured. Pearson’s correlation was used to assess the relationship between leptin and anthropometric parameters while the Mann-Whitney U test was used to assess the differences in cord blood leptin levels between small for gestational age (SGA), appropriate for gestational age (AGA) and LGA infants. There was a significant difference (P < 0.05) between the cord blood leptin concentrations of LGA infants (12.67 ng/mL ± 2.34) and AGA infants (7.10 ng/mL ± 0.90). However, a significant difference was not observed between leptin levels of SGA infants (8.86 ng/mL ± 0.70) and AGA infants. In both male and female neonates, umbilical leptin levels showed significant positive correlations (P < 0.05) with birth weight of the newborn, pre-pregnancy maternal weight and pre pregnancy BMI between the infants of large and appropriate for gestational ages. Increased concentrations of leptin levels in the cord blood of large for gestational age infants suggest that they may be involved in regulating fetal growth. Leptin concentration of Sri Lankan population was not significantly deviated from published data of Asian populations. Fetal leptin may be an important predictor of neonatal adiposity; however, interventional studies are required to assess its impact on the possible risk of childhood obesity.

Keywords: appropriate for gestational age, childhood obesity, leptin, anthropometry

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Authors: Alireza Barari, Ahmad Abdi

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Introduction: Plyometric training (PT) is popular among individuals involved in dynamic sports, and is executed with a goal to improve muscular performance. Cytokines are considered as immunoregulatory molecules for regulation of immune function and other body responses. In addition, the pro-inflammatory cytokines, TNF-α andIL-1β, have been reported to be increased during and after exercises. If some of the cytokines which cause responses such as inflammation of cells in skeletal muscles, with manipulating of training program or optimizing nutrition, it can be avoided or limited from those injuries caused by cytokines release. Its shows that mistletoe extracts show immune-modulating effects. Materials and methods: present study was to investigate the effect of six weeks PT with or without mistletoe supplementation (MS)(10 mg/kg) on cytokine responses and performance in male basketball players. This study is semi-experimental. Statistic society of this study was basketball player’s male students of Mahmoud Abad city. Statistic samples are concluded of 32 basketball players with an age range of 14–17 years was selected from randomly. Selection of samples in four groups of 8 individuals Participants were randomly assigned to either an experimental group (E, n=16) that performed plyometric exercises with (n=8) or without (n=8) MS, or a control group that rested (C, n=16) with (n=8) or without (n=8) MS. Plants were collected in June from the Mazandaran forest in north of Iran. Then they dried in exposure to air without any exposition to sunlight, on a clean textile. For better drying the plants were high and down until they lost their water. Each subject consumed 10 mg/kg/day of extract for six weeks of intervention. Pre and post-testing was performed in the afternoon of the same day. Blood samples (10 ml) were collected from the intermediate cubital vein of the subjects. Serum concentration of IL-1β and TNF-α were measured by ELISA method. Data analysis was performed using pretest to posttest changes that assessed by t-test for paired samples. After the last plyometric training program, the second blood samples were in the next day. Group differences at baseline were evaluated using One-way ANOVA (post-hock Tukey) test is used for analysis and comparison of three group’s variables. Results: PT with or without MS improved the one repetition maximum leg and chest press, Sargeant test and power in RAST (P < 0.05). However there were no statistically significant differences between groups in Vo2max measures (P > 0.05). PT resulted in a significant increase in plasma IL-1β concentration from 1.08±0.4 mg/ml in pre-training to 1.68±0.18 mg/ml in post-training (P=0.006). While the MS significantly decreased the training-induced increment of IL-1β (P=0.007). In contrast, neither PT nor MS had any effect on TNF-α levels (P > 0.05). Discussion: The results of this investigation indicate that PT improved muscular performance and increases the IL-1β concentration. Increasing of IL-1β after exercise in damaged skeletal muscle has shown of the role of this cytokine in inflammation processes and damaged skeletal muscle repair. However mistletoe supplementation ameliorates the increment of IL-1β levels, indicating the beneficial effect of mistletoe on immune response following plyometric training.

Keywords: mistletoe supplementation, training, IL-1β, TNF-α

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Authors: Beatriz González-rodrigo, Diego Hidalgo-leiva, Omar Flores, Claudia Germoso, Maribel Jiménez-martínez, Laura Navas-sánchez, Belén Orta, Nicola Tarque, Orlando Hernández- Rubio, Miguel Marchamalo, Juan Gregorio Rejas, Belén Benito-oterino

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Cities facing seismic hazard necessitate detailed risk assessments for effective urban planning and vulnerability identification, ensuring the safety and sustainability of urban infrastructure. Comprehensive studies involving seismic hazard, vulnerability, and exposure evaluations are pivotal for estimating potential losses and guiding proactive measures against seismic events. However, broad-scale traditional risk studies limit consideration of specific local threats and identify vulnerable housing within a structural typology. Achieving precise results at neighbourhood levels demands higher resolution seismic hazard exposure, and vulnerability studies. This research aims to bolster sustainability and safety against seismic disasters in three Central American and Caribbean capitals. It integrates geospatial techniques and artificial intelligence into seismic risk studies, proposing cost-effective methods for exposure data collection and damage prediction. The methodology relies on prior seismic threat studies in pilot zones, utilizing existing exposure and vulnerability data in the region. Emphasizing detailed building attributes enables the consideration of behaviour modifiers affecting seismic response. The approach aims to generate detailed risk scenarios, facilitating prioritization of preventive actions pre-, during, and post-seismic events, enhancing decision-making certainty. Detailed risk scenarios necessitate substantial investment in fieldwork, training, research, and methodology development. Regional cooperation becomes crucial given similar seismic threats, urban planning, and construction systems among involved countries. The outcomes hold significance for emergency planning and national and regional construction regulations. The success of this methodology depends on cooperation, investment, and innovative approaches, offering insights and lessons applicable to regions facing moderate seismic threats with vulnerable constructions. Thus, this framework aims to fortify resilience in seismic-prone areas and serves as a reference for global urban planning and disaster management strategies. In conclusion, this research proposes a comprehensive framework for seismic risk assessment in high-risk urban areas, emphasizing detailed studies at finer resolutions for precise vulnerability evaluations. The approach integrates regional cooperation, geospatial technologies, and adaptive fragility curve adjustments to enhance risk assessment accuracy, guiding effective mitigation strategies and emergency management plans.

Keywords: assessment, behaviour modifiers, emergency management, mitigation strategies, resilience, vulnerability

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Authors: Ana L. Sánchez Regla, Igor A. Rivera González, María del Pilar Monserrat Pérez Hernández

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The ability of an organization to identify and get useful information from external sources, assimilate it, transform and apply to generate products or services with added value is called absorptive capacity. Absorptive capabilities contribute to have market opportunities to firms and get a leader position with respect to others competitors. The Bioprocess Development and Research Unit (UDIBI) is a Research and Development (R&D) laboratory that belongs to the National Polytechnic Institute (IPN), which is a higher education institute in Mexico. The UDIBI was created with the purpose of carrying out R and D activities for the Transferon®, a biopharmaceutical product developed and patented by IPN. The evolution of competence and scientific and technological platform made UDIBI expand its scope by providing technological services (preclínical studies and bio-compatibility evaluation) to the national pharmaceutical industry and biopharmaceutical industry. The relevance of this study is that those industries are classified as high scientific and technological intensity, and yet, after a review of the state of the art, there is only one study of absorption capabilities in biopharmaceutical industry with a similar scope to this research; in the case of Mexico, there is none. In addition to this, UDIBI belongs to a public university and its operation does not depend on the federal budget, but on the income generated by its external technological services. This fact represents a highly remarkable case in Mexico's public higher education context. This current doctoral research (2015-2019) is contextualized within a case study, its main objective is to identify and analyze the absorptive capabilities that characterise the UDIBI that allows it had become in a one of two third authorized laboratory by the sanitary authority in Mexico for developed bio-comparability studies to bio-pharmaceutical products. The development of this work in the field is divided into two phases. In a first phase, 15 interviews were conducted with the UDIBI personnel, covering management levels, heads of services, project leaders and laboratory personnel. These interviews were structured under a questionnaire, which was designed to integrate open questions and to a lesser extent, others, whose answers would be answered on a Likert-type rating scale. From the information obtained in this phase, a scientific article was made (in review and a proposal of presentation was submitted in different academic forums. A second stage will be made from the conduct of an ethnographic study within this organization under study that will last about 3 months. On the other hand, it is intended to carry out interviews with external actors around the UDIBI (suppliers, advisors, IPN officials, including contact with an academic specialized in absorption capacities to express their comments on this thesis. The inicial findings had shown two lines: i) exist institutional, technological and organizational management elements that encourage and/or limit the creation of absorption capacities in this scientific and technological laboratory and, ii) UDIBI has had created a set of multiple transfer technology of knowledge mechanisms which have had permitted to build a huge base of prior knowledge.

Keywords: absorptive capabilities, biopharmaceutical industry, high research and development intensity industries, knowledge management, transfer of knowledge

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Authors: Gachara Grace, Kebira Anthony, Harvey Jagger, Wainaina James

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Aflatoxin contamination of maize in Africa poses a major threat to food security and the health of many African people. In Kenya, aflatoxin contamination of maize is high due to the environmental, agricultural and socio-economic factors. Many studies have been conducted to understand the scope of the problem, especially at pre-harvest level. This research was carried out to gather scientific information on the fungi population, diversity and aflatoxin level during the post-harvest period. The study was conducted in three geographical locations of; Kitui, Kitale and Nakuru. Samples were collected from storage structures of farmers and transported to the Biosciences eastern and central Africa (BecA), International Livestock and Research Institute (ILRI) hub laboratories. Mycoflora was recovered using the direct plating method. A total of five fungal genera (Aspergillus, Penicillium, Fusarium, Rhizopus and Bssyochlamys spp.) were isolated from the stored maize samples. The most common fungal species that were isolated from the three study sites included A. flavus at 82.03% followed by A.niger and F.solani at 49% and 26% respectively. The aflatoxin producing fungi A. flavus was recovered in 82.03% of the samples. Aflatoxin levels were analysed on both the maize samples and in vitro. Most of the A. flavus isolates recorded a high level of aflatoxin when they were analysed for presence of aflatoxin B1 using ELISA. In Kitui, all the samples (100%) had aflatoxin levels above 10ppb with a total aflatoxin mean of 219.2ppb. In Kitale, only 3 samples (n=39) had their aflatoxin levels less than 10ppb while in Nakuru, the total aflatoxin mean level of this region was 239.7ppb. When individual samples were analysed using Vicam fluorometer method, aflatoxin analysis revealed that most of the samples (58.4%) had been contaminated. The means were significantly different (p=0.00<0.05) in all the three locations. Genetic relationships of A. flavus isolates were determined using 13 Simple Sequence Repeats (SSRs) markers. The results were used to generate a phylogenetic tree using DARwin5 software program. A total of 5 distinct clusters were revealed among the genotypes. The isolates appeared to cluster separately according to the geographical locations. Principal Coordinates Analysis (PCoA) of the genetic distances among the 91 A. flavus isolates explained over 50.3% of the total variation when two coordinates were used to cluster the isolates. Analysis of Molecular Variance (AMOVA) showed a high variation of 87% within populations and 13% among populations. This research has shown that A. flavus is the main fungal species infecting maize grains in Kenya. The influence of aflatoxins on human populations in Kenya demonstrates a clear need for tools to manage contamination of locally produced maize. Food basket surveys for aflatoxin contamination should be conducted on a regular basis. This would assist in obtaining reliable data on aflatoxin incidence in different food crops. This would go a long way in defining control strategies for this menace.

Keywords: aflatoxin, Aspergillus flavus, genotyping, Kenya

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Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry, because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 6 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility of using this biosensor to address the challenge associated with pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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Authors: Margarita Ivanova, Julia Dao, Andrew Friedman, Neil Kasaci, Rekha Gopal, Ozlem Goker-Alpan

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In Fabry disease (FD), α-galactosidase A (α-Gal A) deficiency leads to the accumulation of globotriaosylceramide (Lyso-Gb3 and Gb3), triggering a pathologic cascade that causes the severity of organs damage. The heart is one of the several organs with high sensitivity to the α-Gal A deficiency. A subgroup of patients with significant residual of α-Gal A activity with primary cardiac involvement is occasionally referred to as “cardiac variant.” The cardiovascular complications are most frequently encountered, contributing substantially to morbidity, and are the leading cause of premature death in male and female patients with FD. The deposition of Lyso-Gb-3 and Gb-3 within the myocardium affects cardiac function with resultant progressive cardiovascular pathology. Gb-3 and Lyso-Gb-3 accumulation at the cellular level trigger a cascade of events leading to end-stage fibrosis. In the cardiac tissue, Lyso-Gb-3 deposition is associated with the increased release of inflammatory factors and transforming growth factors. Infiltration of lymphocytes and macrophages into endomyocardial tissue indicates that inflammation plays a significant role in cardiac damage. Moreover, accumulated data suggest that chronic inflammation leads to multisystemic FD pathology even under enzyme replacement therapy (ERT). NF-κB activation plays a subsequent role in the inflammatory response to cardiac dysfunction and advanced heart failure in the general population. TNFalpha/NF-κB signaling protects the myocardial evoking by ischemic preconditioning; however, this protective effect depends on the concentration of TNF-α. Thus, we hypothesize that TNF-α is a critical factor in determining the grade of cardio-pathology. Cardiac hypertrophy corresponds to the expansion of the coronary vasculature to maintain a sufficient supply of nutrients and oxygen. Coronary activation of angiogenesis and fibrosis plays a vital role in cardiac vascularization, hypertrophy, and tissue remodeling. We suggest that the interaction between the inflammatory pathways and cardiac vascularization is a bi-directional process controlled by secreted cytokines and growth factors. The co-coordination of these two processes has never been explored in FD. In a cohort of 40 patients with FD, biomarkers associated with inflammation and cardio hypertrophic remodeling were studied. FD patients were categorized into three groups based on LVmass/DSA, LVEF, and ECG abnormalities: FD with no cardio complication, FD with moderate cardio complication, and severe cardio complication. Serum levels of NF-kB, TNFalpha, Il-6, Il-2, MCP1, ING-gamma, VEGF, IGF-1, TGFβ, and FGF2 were quantified by enzyme-linked immunosorbent assays (ELISA). Among the biomarkers, MCP-1, INF-gamma, VEGF, TNF-alpha, and TGF-beta were elevated in FD patients. Some of these biomarkers also have the potential to correlate with cardio pathology in FD. Conclusion: The study provides information about the role of inflammatory pathways and biomarkers of cardio hypertrophic remodeling in FD patients. This study will also reveal the mechanisms that link intracellular accumulation of Lyso-GB-3 and Gb3 to the development of cardiomyopathy with myocardial thickening and resultant fibrosis.

Keywords: biomarkers, Fabry disease, inflammation, growth factors

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Authors: Esra Sengul, R. G. Aktas, M. E. Sitar, H. Isan

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Hepatocellular carcinoma (HCC) is a hepatocellular tumor commonly found on the surface of the chronic liver. HepG2 is the most commonly used cell type in HCC studies. The main proteins remaining in the blood serum after separation of plasma fibrinogen are albumin and globulin. The fact that the albumin showed hepatocellular damage and reflect the synthesis capacity of the liver was the main reason for our use. Alpha-Fetoprotein (AFP) is an albumin-like structural embryonic globulin found in the embryonic cortex, cord blood, and fetal liver. It has been used as a marker in the follow-up of tumor growth in various malign tumors and in the efficacy of surgical-medical treatments, so it is a good protein to look at with albumins. We have seen the morphological changes of dimethyl sulfoxide (DMSO) on HepG2 and decided to investigate its biochemical effects. We examined the effects of DMSO, which is used in cell cultures, on albumin, AFP and total protein at low doses. Material Method: Cell Culture: Medium was prepared in cell culture using Dulbecco's Modified Eagle Media (DMEM), Fetal Bovine Serum Dulbecco's (FBS), Phosphate Buffered Saline and trypsin maintained at -20 ° C. Fixation of Cells: HepG2 cells, which have been appropriately developed at the end of the first week, were fixed with acetone. We stored our cells in PBS at + 4 ° C until the fixation was completed. Area Calculation: The areas of the cells are calculated in the ImageJ (IJ). Microscope examination: The examination was performed with a Zeiss Inverted Microscope. Daytime photographs were taken at 40x, 100x 200x and 400x. Biochemical Tests: Protein (Total): Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Albumin: Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Alpha-fetoprotein: Serum sample was analyzed by ECLIA method. Results: When liver cancer cells were cultured in medium with 1% DMSO for 4 weeks, a significant difference was observed when compared with the control group. As a result, we have seen that DMSO can be used as an important agent in the treatment of liver cancer. Cell areas were reduced in the DMSO group compared to the control group and the confluency ratio increased. The ability to form spheroids was also significantly higher in the DMSO group. Alpha-fetoprotein was lower than the values of an ordinary liver cancer patient and the total protein amount increased to the reference range of the normal individual. Because the albumin sample was below the specimen value, the numerical results could not be obtained on biochemical examinations. We interpret all these results as making DMSO a caretaking aid. Since each one was not enough alone we used 3 parameters and the results were positive when we refer to the values of a normal healthy individual in parallel. We hope to extend the study further by adding new parameters and genetic analyzes, by increasing the number of samples, and by using DMSO as an adjunct agent in the treatment of liver cancer.

Keywords: hepatocellular carcinoma, HepG2, dimethyl sulfoxide, cell culture, ELISA

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Authors: Kunal Garg, Louise Theusen Hermansan, Kanoktip Puttaraska, Oliver Hendricks, Heidi Pirttinen, Leona Gilbert

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The Germ Theory (one infectious determinant is equal to one disease) has unarguably evolved our capability to diagnose and treat infectious diseases over the years. Nevertheless, the advent of technology, climate change, and volatile human behavior has brought about drastic changes in our environment, leading us to question the relevance of the Germ Theory in our day, i.e. will vector-borne disease (VBD) sufferers produce multiple immune responses when tested for multiple microbes? Vector diseased patients producing multiple immune responses to different microbes would evidently suggest human polymicrobial infections (HPI). Ongoing diagnostic tools are exceedingly unequipped with the current research findings that would aid in diagnosing patients for polymicrobial infections. This shortcoming has caused misdiagnosis at very high rates, consequently diminishing the patient’s quality of life due to inadequate treatment. Equipped with the state-of-art scientific knowledge, PolyScan intends to address the pitfalls in current VBD diagnostics. PolyScan is a multiplex and multifunctional enzyme linked Immunosorbent assay (ELISA) platform that can test for numerous VBD microbes and allow simultaneous screening for multiple types of antibodies. To validate PolyScan, Lyme Borreliosis (LB) and spondyloarthritis (SpA) patient groups (n = 54 each) were tested for Borrelia burgdorferi, Borrelia burgdorferi Round Body (RB), Borrelia afzelii, Borrelia garinii, and Ehrlichia chaffeensis against IgM and IgG antibodies. LB serum samples were obtained from Germany and SpA serum samples were obtained from Denmark under relevant ethical approvals. The SpA group represented chronic LB stage because reactive arthritis (SpA subtype) in the form of Lyme arthritis links to LB. It was hypothesized that patients from both the groups will produce multiple immune responses that as a consequence would evidently suggest HPI. It was also hypothesized that the multiple immune response proportion in SpA patient group would be significantly larger when compared to the LB patient group across both antibodies. It was observed that 26% LB patients and 57% SpA patients produced multiple immune responses in contrast to 33% LB patients and 30% SpA patients that produced solitary immune responses when tested against IgM. Similarly, 52% LB patients and an astounding 73% SpA patients produced multiple immune responses in contrast to 30% LB patients and 8% SpA patients that produced solitary immune responses when tested against IgG. Interestingly, IgM immune dysfunction in both the patient groups was also recorded. Atypically, 6% of the unresponsive 18% LB with IgG antibody was recorded producing multiple immune responses with the IgM antibody. Similarly, 12% of the unresponsive 19% SpA with IgG antibody was recorded producing multiple immune responses with the IgM antibody. Thus, results not only supported hypothesis but also suggested that IgM may atypically prevail longer than IgG. The PolyScan concept will aid clinicians to detect patients for early, persistent, late, polymicrobial, & immune dysfunction conditions linked to different VBD. PolyScan provides a paradigm shift for the VBD diagnostic industry to follow that will drastically shorten patient’s time to receive adequate treatment.

Keywords: diagnostics, immune dysfunction, polymicrobial, TICK-TAG

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Authors: Olugbenga S. Alebiosu, Olusola H. Adekanmbi, Oluwatoyin T. Ogundipe

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Pollen and spores have been identified as major airborne bio-particles inducing respiratory disorders such as asthma, allergic rhinitis and atopic dermatitis among hypersensitive individuals. An aeropalynological study was conducted within a one year sampling period with a view to investigating the monthly depositional rate of atmospheric pollen and spores; influence of the immediate vegetation on airborne pollen distribution; allergenic potentials of dominant atmospheric pollen types at selected study locations in Bauchi and Taraba states, Northwestern Nigeria. A tauber-like pollen trap was employed in aerosampling with the sampler positioned at a height of 5 feet above the ground, followed by a monthly collection of the recipient solution for the sampling period. The collected samples were subjected to acetolysis treatment, examined microscopically with the identification of pollen grains and spores using reference materials and published photomicrographs. Plants within the surrounding vegetation were enumerated. Crude protein contents extracted from pollen types found to be commonly dominant at both study locations; Senna siamea, Terminalia cattapa, Panicum maximum and Zea mays were used to sensitize Musmusculus. Histopathological studies of bronchi and lung sections from certain dead M.musculus in the test groups was conducted. Blood samples were collected from the pre-orbital vein of M.musculus and processed for serological and haematological (differential and total white blood cell counts) studies. ELISA was used in determining the levels of serological parameters: IgE and cytokines (TNF-, IL-5, and IL-13). Statistical significance was observed in the correlation between the levels of serological and haematological parameters elicited by each test group, differences between the levels of serological and haematological parameters elicited by each test group and those of the control, as well as at varying sensitization periods. The results from this study revealed dominant airborne pollen types across the study locations; Syzygiumguineense, Tridaxprocumbens, Elaeisguineensis, Mimosa sp., Borreria sp., Terminalia sp., Senna sp. and Poaceae. Nephrolepis sp., Pteris sp. and a trilete fern also produced spores. This study also revealed that some of the airborne pollen types were produced by local plants at the study locations. Bronchi sections of M.musculus after first and second sensitizations, as well as lung section after first sensitization with Senna siamea, showed areas of necrosis. Statistical significance was recorded in the correlation between the levels of some serological and haematological parameters produced by each test group and those of the control, as well as at certain sensitization periods. The study revealed some candidate pollen allergens at the study locations allergy sufferers and also established a complexity of interaction between immune cells, IgE and cytokines at varied periods of mice sensitization and forming a paradigm of human immune response to different pollen allergens. However, it is expedient that further studies should be conducted on these candidate pollen allergens for their allergenicity potential in humans within their immediate environment.

Keywords: airborne, hypersensitive, mus musculus, pollen allergens, respiratory, tauber-like

Procedia PDF Downloads 109

Authors: Hongyan Chen, Megan Jobson, Andrew J. Masters, Maria Gonzalez-Miquel, Simon Halstead, Mayri Diaz de Rienzo

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Ionic liquids offer excellent advantages over conventional solvents for industrial extraction of metals from aqueous solutions, where such extraction processes bring opportunities for recovery, reuse, and recycling of valuable resources and more sustainable production pathways. Recent research on the use of ionic liquids for extraction confirms their high selectivity and low volatility, but there is relatively little focus on how their properties can be best exploited in practice. This work addresses gaps in research on process modelling and simulation, to support development, design, and optimisation of these processes, focusing on the separation of the highly similar transition metals, cobalt, and nickel. The study exploits published experimental results, as well as new experimental results, relating to the separation of Co and Ni using trihexyl (tetradecyl) phosphonium chloride. This extraction agent is attractive because it is cheaper, more stable and less toxic than fluorinated hydrophobic ionic liquids. This process modelling work concerns selection and/or development of suitable models for the physical properties, distribution coefficients, for mass transfer phenomena, of the extractor unit and of the multi-stage extraction flowsheet. The distribution coefficient model for cobalt and HCl represents an anion exchange mechanism, supported by the literature and COSMO-RS calculations. Parameters of the distribution coefficient models are estimated by fitting the model to published experimental extraction equilibrium results. The mass transfer model applies Newman’s hard sphere model. Diffusion coefficients in the aqueous phase are obtained from the literature, while diffusion coefficients in the ionic liquid phase are fitted to dynamic experimental results. The mass transfer area is calculated from the surface to mean diameter of liquid droplets of the dispersed phase, estimated from the Weber number inside the extractor. New experiments measure the interfacial tension between the aqueous and ionic phases. The empirical models for predicting the density and viscosity of solutions under different metal loadings are also fitted to new experimental data. The extractor is modelled as a continuous stirred tank reactor with mass transfer between the two phases and perfect phase separation of the outlet flows. A multistage separation flowsheet simulation is set up to replicate a published experiment and compare model predictions with the experimental results. This simulation model is implemented in gPROMS software for dynamic process simulation. The results of single stage and multi-stage flowsheet simulations are shown to be in good agreement with the published experimental results. The estimated diffusion coefficient of cobalt in the ionic liquid phase is in reasonable agreement with published data for the diffusion coefficients of various metals in this ionic liquid. A sensitivity study with this simulation model demonstrates the usefulness of the models for process design. The simulation approach has potential to be extended to account for other metals, acids, and solvents for process development, design, and optimisation of extraction processes applying ionic liquids for metals separations, although a lack of experimental data is currently limiting the accuracy of models within the whole framework. Future work will focus on process development more generally and on extractive separation of rare earths using ionic liquids.

Keywords: distribution coefficient, mass transfer, COSMO-RS, flowsheet simulation, phosphonium

Procedia PDF Downloads 154

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

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

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

Procedia PDF Downloads 130

Authors: Fernanda Piculo, Giovana Vesentini, Gabriela Marini, Debora Cristina Damasceno, Angelica Mercia Pascon Barbosa, Marilza Vieira Cunha Rudge

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Introduction: Women with previous gestational diabetes mellitus (GDM) were significantly more likely to have urinary incontinence (UI) and pelvic floor muscle dysfunction compared to non-diabetic women two years after a cesarean section. Additional results demonstrated that induced diabetes causes detrimental effects on pregnant rat urethral muscle. These results indicate the need for exploration of the mechanistic role of a recovery factor in female UI. Chemokine ligand 7 (CCL7) was significantly over expressed in rat serum, urethral and vaginal tissues immediately following induction of stress UI in a rat model simulating birth trauma. CCL7 over expression has shown potency for stimulating targeted stem cell migration and provide a translational link (clinical measurement) which further provide opportunities for treatment. The aim of this study was to investigate the CCL7 levels profile in diabetic pregnant women with urinary incontinence during pregnancy over the first year postpartum. Methods: This study was conducted in the Perinatal Diabetes Research Center of the Botucatu Medical School/UNESP, and was approved by the Research Ethics Committee of the Institution (CAAE: 20639813.0.0000.5411). The diagnosis of GDM was established between 24th and 28th gestational weeks, by the 75 g-OGTT test according to ADA’s criteria. Urinary incontinence was defined according to the International Continence Society and the CCL7 levels was measured by ELISA (R&D Systems, Catalog Number DCC700). Two hundred twelve women were classified into four study groups: normoglycemic continent (NC), normoglycemic incontinent (NI), diabetic continent (DC) and diabetic incontinent (DI). They were evaluated at six-time-points: 12-18, 24-28 and 34-38 gestational weeks, 24-48 hours, 6 weeks and 6-12 months postpartum. Results: At 12-18 weeks, it was possible to consider only two groups, continent and incontinent, because at this early gestational period has not yet been the diagnosis of GDM. The group with GDM and UI (DI group) showed lower levels of CCL7 in all time points during pregnancy and postpartum, compared to normoglycemic groups (NC and NI), indicating that these women have not recovered from child birth induced UI during the 6-12 months postpartum compared to their controls, and that the progression of UI and/or lack of recovery throughout the first postpartum year can be related with lower levels of CCL7. Instead, serum CCL7 was significantly increased in the NC group. Taken together, these findings of overexpression of CCL7 in the NC group and decreased levels in the DI group, could confirm that diabetes delays the recovery from child birth induced UI, and that CCL7 could potentially be used as a serum marker of injury. Conclusion: This study demonstrates lower levels of CCL7 in the DI group during pregnancy and postpartum and suggests that the progression of UI in diabetic women and/or lack of recovery throughout the first postpartum year can be related with low levels of CCL7. This provides a translational potential where CCL7 measurement could be used as a surrogate for injury after delivery. Successful controlled CCL7 mediated stem cell homing to the lower urinary tract could one day introduce the potential for non-operative treatment or prevention of stress urinary incontinence.

Keywords: CCL7, gestational diabetes, pregnancy, urinary incontinence

Procedia PDF Downloads 308

Authors: Alireza Barari, Saeed Shirali, Ahmad Abdi

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Abstract: Introduction: Physical activity may be related to male reproductive function by affecting on thehypothalamic-pituitary-gonadal(HPG) axis. Our aim was to determine the effects of 6 weeks resistance exercise on reproductive hormones, HPG axis. The hypothalamic-pituitary-gonadal (HPG) axis refers tothe effects of endocrine glands in three-level including (i) the hypothalamic releasing hormone GnRH, which is synthesized in in a small heterogenous neuronal population and released in a pulsatile fashion, (ii) the anterior pituitary hormones, follicle-stimulating hormone(FSH) and luteinizing hormone (LH) and (iii) the gonadal hormones, which include both steroid such as testosterone (T), estradiol and progesterone and peptide hormones (such as inhibin). Hormonal changes that create a more anabolic environment have been suggested to contribute to the adaptation to strength exercise. Physical activity has an extensive impact on male reproductive function depending upon the intensity and duration of the exercise and the fitness level of the individual. However, strenuous exercise represents a physical stress and inflammation changed that challenges homeostasis. Materials and methods: Sixteen male volunteered were included in a 6-week control period followed by 6 weeks of resistance training (leg press, lat pull, chest press, squat, seatedrow, abdominal crunch, shoulder press, biceps curl and triceps press down) four times per week. intensity of training loading was 60%-75% of one maximum repetition. Participants performed 3 sets of 10 repetitions. Rest periods were two min between exercises and sets. Start with warm up exercises include: The muscles relax and stretch the body, which was for 10 minutes. Body composition, VO2max and the circulating level of free testosterone (fT), luteinizing hormone (LH), follicle-stimulating hormone (FSH), sex hormone binding globulin (SHBG) and inhibin B measured prior and post 6-week intervention. The hormonal levels of each serum sample were measured using commercially available ELISA kits. Analysis of anthropometrical data and hormonal level were compared using the independent samples t- test in both groups and using SPSS (version 19). P ≤ 0.05 was considered statistically significant. Results: For muscle strength, both lower- and upper-body strength were increased significantly. Aerobic fitness level improved in trained participant from 39.4 ± 5.6 to 41.9 ± 5.3 (P = 0.002). fT concentration rise progressively in the trained group and was significantly greater than those in the control group (P = 0.000). By the end of the 6-week resistance training, serum SHBG significantly increased in the trained group compared with the control group (P = 0.013). In response to resistance training, LH, FSH and inhibin B were not significantly changed. Discussion: According to our finfings, 6 weeks of resistance training induce fat loss without any changes in body weight and BMI. A decline of 25.3% in percentage of body fat with statiscally same weight was due to increase in muscle mass that happened during resistance exercise periods . Six weeks of resistance training resulted in significant improvement in BF%, VO2max and increasing strength and the level of fT and SHBG.

Keywords: resistance, hypothalamic, pituitary, gonadal axis

Procedia PDF Downloads 375

Authors: Elisa Menardo, Margherita Pasini, Margherita Brondino

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A growing body of empirical research from different areas of inquiry suggests that brief contact with natural environment restore mental resources. The Attention Restoration Theory (ART) is the widespread used and empirical founded theory developed to explain why exposure to nature helps people to recovery cognitive resources. It assumes that contact with nature allows people to free (and then recovery) voluntary attention resources and thus allows them to recover from a cognitive fatigue situation. However, it was suggested that some people could have more cognitive benefit after exposure to urban environment. The objective of this study is to report the results of a meta-analysis on studies (peer-reviewed articles) comparing the restorativeness (the quality to be restorative) perceived in natural environments than those perceived in urban environments. This meta-analysis intended to estimate how much nature environments (forests, parks, boulevards) are perceived to be more restorativeness than urban ones (i.e., the magnitude of the perceived restorativeness’ difference). Moreover, given the methodological difference between study, it studied the potential role of moderator variables as participants (student or other), instrument used (Perceived Restorativeness Scale or other), and procedure (in laboratory or in situ). PsycINFO, PsycARTICLES, Scopus, SpringerLINK, Web of Science online database were used to identify all peer-review articles on restorativeness published to date (k = 167). Reference sections of obtained papers were examined for additional studies. Only 22 independent studies (with a total of 1371 participants) met inclusion criteria (direct exposure to environment, comparison between one outdoor environment with natural element and one without natural element, and restorativeness measured by self-report scale) and were included in meta-analysis. To estimate the average effect size, a random effect model (Restricted Maximum-likelihood estimator) was used because the studies included in the meta-analysis were conducted independently and using different methods in different populations, so no common effect-size was expected. The presence of publication bias was checked using trim and fill approach. Univariate moderator analysis (mixed effect model) were run to determine whether the variable coded moderated the perceived restorativeness difference. Results show that natural environments are perceived to be more restorativeness than urban environments, confirming from an empirical point of view what is now considered a knowledge gained in environmental psychology. The relevant information emerging from this study is the magnitude of the estimated average effect size, which is particularly high (d = 1.99) compared to those that are commonly observed in psychology. Significant heterogeneity between study was found (Q(19) = 503.16, p < 0.001;) and studies’ variability was very high (I2[C.I.] = 96.97% [94.61 - 98.62]). Subsequent univariate moderator analyses were not significant. Methodological difference (participants, instrument, and procedure) did not explain variability between study. Other methodological difference (e.g., research design, environment’s characteristics, light’s condition) could explain this variability between study. In the mine while, studies’ variability could be not due to methodological difference but to individual difference (age, gender, education level) and characteristics (connection to nature, environmental attitude). Furthers moderator analysis are working in progress.

Keywords: meta-analysis, natural environments, perceived restorativeness, urban environments

Procedia PDF Downloads 144

Authors: Katarzyna Lubecka, Kirsty Flower, Megan Beetch, Lucinda Kurzava, Hannah Buvala, Samer Gawrieh, Suthat Liangpunsakul, Tracy Gonzalez, George McCabe, Naga Chalasani, James M. Flanagan, Barbara Stefanska

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Hepatocellular carcinoma (HCC), the most prevalent type of primary liver cancer, is the second leading cause of cancer death worldwide. Late onset of clinical symptoms in HCC results in late diagnosis and poor disease outcome. Approximately 85% of individuals with HCC have underlying liver cirrhosis. However, not all cirrhotic patients develop cancer. Reliable early detection biomarkers that can distinguish cirrhotic patients who will develop cancer from those who will not are urgently needed and could increase the cure rate from 5% to 80%. We used Illumina-450K microarray to test whether blood DNA, an easily accessible source of DNA, bear site-specific changes in DNA methylation in response to HCC before diagnosis with conventional tools (pre-diagnostic). Top 11 differentially methylated sites were selected for validation by pyrosequencing. The diagnostic potential of the 11 pyrosequenced probes was tested in blood samples from a prospective cohort of cirrhotic patients. We identified 971 differentially methylated CpG sites in pre-diagnostic HCC cases as compared with healthy controls (P < 0.05, paired Wilcoxon test, ICC ≥ 0.5). Nearly 76% of differentially methylated CpG sites showed lower levels of methylation in cases vs. controls (P = 2.973E-11, Wilcoxon test). Classification of the CpG sites according to their location relative to CpG islands and transcription start site revealed that those hypomethylated loci are located in regulatory regions important for gene transcription such as CpG island shores, promoters, and 5’UTR at higher frequency than hypermethylated sites. Among 735 CpG sites hypomethylated in cases vs. controls, 482 sites were assigned to gene coding regions whereas 236 hypermethylated sites corresponded to 160 genes. Bioinformatics analysis using GO, KEGG and DAVID knowledgebase indicate that differentially methylated CpG sites are located in genes associated with functions that are essential for gene transcription, cell adhesion, cell migration, and regulation of signal transduction pathways. Taking into account the magnitude of the difference, statistical significance, location, and consistency across the majority of matched pairs case-control, we selected 11 CpG loci corresponding to 10 genes for further validation by pyrosequencing. We established that methylation of CpG sites within 5 out of those 10 genes distinguish cirrhotic patients who subsequently developed HCC from those who stayed cancer free (cirrhotic controls), demonstrating potential as biomarkers of early detection in populations at risk. The best predictive value was detected for CpGs located within BARD1 (AUC=0.70, asymptotic significance ˂0.01). Using an additive logistic regression model, we further showed that 9 CpG loci within those 5 genes, that were covered in pyrosequenced probes, constitute a panel with high diagnostic accuracy (AUC=0.887; 95% CI:0.80-0.98). The panel was able to distinguish pre-diagnostic cases from cirrhotic controls free of cancer with 88% sensitivity at 70% specificity. Using blood as a minimally invasive material and pyrosequencing as a straightforward quantitative method, the established biomarker panel has high potential to be developed into a routine clinical test after validation in larger cohorts. This study was supported by Showalter Trust, American Cancer Society (IRG#14-190-56), and Purdue Center for Cancer Research (P30 CA023168) granted to BS.

Keywords: biomarker, DNA methylation, early detection, hepatocellular carcinoma

Procedia PDF Downloads 267

Authors: Rijnbout-St.James Willem, Lindner Volkhard, Scholand Mary Beth, Ashton M. Tillett, Di Gennaro Michael Jude, Smith Silvia Enrica

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Introduction: Fibrosing lung diseases are characterized by changes in the lung interstitium and are classified based on etiology: 1) environmental/exposure-related, 2) autoimmune-related, 3) sarcoidosis, 4) interstitial pneumonia, and 4) idiopathic. Among interstitial lung diseases (ILD) idiopathic forms, idiopathic pulmonary fibrosis (IPF) is the most severe. Pathogenesis of IPF is characterized by an increased presence of proinflammatory mediators, resulting in alveolar injury, where injury to alveolar epithelium precipitates an increase in collagen deposition, subsequently thickening the alveolar septum and decreasing gas exchange. Identifying biomarkers implicated in the pathogenesis of lung fibrosis is key to developing new therapies and improving the efficacy of existing therapies. The transforming growth factor-beta (TGF-B1), a mediator of tissue repair associated with WNT5A signaling, is partially responsible for fibroblast proliferation in ILD and is the target of Pirfenidone, one of the antifibrotic therapies used for patients with IPF. Canonical TGF-B signaling is mediated by the proteins SMAD 2/3, which are, in turn, indirectly regulated by Collagen Triple Helix Repeat Containing 1 (CTHRC1). In this study, we tested the following hypotheses: 1) CTHRC1 is more elevated in the ILD cohort compared to unaffected controls, and 2) CTHRC1 is differently expressed among ILD types. Material and Methods: CTHRC1 levels were measured by ELISA in 171 plasma samples from the deidentified University of Utah ILD cohort. Data represent a cohort of 131 ILD-affected participants and 40 unaffected controls. CTHRC1 samples were categorized by a pulmonologist based on affectation status and disease subtypes: IPF (n = 45), sarcoidosis (4), nonspecific interstitial pneumonia (16), hypersensitivity pneumonitis (n = 7), interstitial pneumonia (n=13), autoimmune (n = 15), other ILD - a category that includes undifferentiated ILD diagnoses (n = 31), and unaffected controls (n = 40). We conducted a single-factor ANOVA of plasma CTHRC1 levels to test whether CTHRC1 variance among affected and non-affected participants is statistically significantly different. In-silico analysis was performed with Ingenuity Pathway Analysis® to characterize the role of CTHRC1 in the pathway of lung fibrosis. Results: Statistical analyses of CTHRC1 in plasma samples indicate that the average CTHRC1 level is significantly higher in ILD-affected participants than controls, with the autoimmune ILD being higher than other ILD types, thus supporting our hypotheses. In-silico analyses show that CTHRC1 indirectly activates and phosphorylates SMAD3, which in turn cross-regulates TGF-B1. CTHRC1 also may regulate the expression and transcription of TGFB-1 via WNT5A and its regulatory relationship with CTNNB1. Conclusion: In-silico pathway analyses demonstrate that CTHRC1 may be an important biomarker in ILD. Analysis of plasma samples indicates that CTHRC1 expression is positively associated with ILD affectation, with autoimmune ILD having the highest average CTHRC1 values. While characterizing CTHRC1 levels in plasma can help to differentiate among ILD types and predict response to Pirfenidone, the extent to which plasma CTHRC1 level is a function of ILD severity or chronicity is unknown.

Keywords: interstitial lung disease, CTHRC1, idiopathic pulmonary fibrosis, pathway analyses

Procedia PDF Downloads 163

Authors: M. Skibinska, A. Rajewska-Rager, M. Dmitrzak-Weglarz, N. Lepczynska, P. Sibilski, P. Kapelski, J. Pawlak, J. Twarowska-Hauser

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Introduction: Neurotrophic factors have been implicated in neuropsychiatric disorders. Brain-Derived Neurotrophic Factor (BDNF) influences neuron differentiation in development as well as synaptic plasticity and neuron survival in adulthood. BDNF is widely studied in mood disorders and has been proposed as a biomarker for depression. BDNF is synthesized as precursor protein – proBDNF. Both forms are biologically active and exert opposite effects on neurons. Aim: The aim of the study was to examine the serum levels of BDNF and proBDNF in unipolar and bipolar young patients below 24 years old during hypo/manic, depressive episodes and in remission compared to healthy control group. Methods: In a prospective 2 years follow-up study, we investigated alterations in levels of BDNF and proBDNF in 79 patients (23 males, mean age 19.08, SD 3.3 and 56 females, mean age 18.39, SD 3.28) diagnosed with mood disorders: unipolar and bipolar disorder compared with 35 healthy control subjects (7 males, mean age 20.43, SD 4.23 and 28 females, mean age 21.25, SD 2.11). Clinical characteristics including mood, comorbidity, family history, and treatment, were evaluated during control visits and clinical symptoms were rated using the Hamilton Depression Rating Scale and Young Mania Rating Scale. Serum BDNF and proBDNF concentrations were determined by Enzyme-Linked Immunosorbent Assays (ELISA) method. Serum BDNF and proBDNF levels were analysed with covariates: sex, age, age > 18 and < 18 years old, family history of affective disorders, drug-free vs. medicated status. Normality of the data was tested using Shapiro-Wilk test. Levene’s test was used to calculate homogeneity of variance. Non-parametric Tests: Mann-Whitney U test, Kruskal-Wallis ANOVA, Friedman’s ANOVA, Wilcoxon signed rank test, Spearman correlation coefficient were applied in analyses The statistical significance level was set at p < 0.05. Results: BDNF and proBDNF serum levels did not differ between patients at baseline and controls as well as comparing patients in acute episode of depression/hypo/mania at baseline and euthymia (at month 3 or 6). Comparing BDNF and proBDNF levels between patients in euthymia and control group no differences have been found. Increased BDNF level in women compared to men at baseline (p=0.01) have been observed. BDNF level at baseline was negatively correlated with depression and mania occurence at 24 month (p=0.04). BDNF level at 12 month was negatively correlated with depression and mania occurence at 12 month (p=0.01). Correlation of BDNF level with sex have been detected (p=0.01). proBDNF levels at month 3, 6 and 12 negatively correlated with disease status (p=0.02, p=0.008, p=0.009, respectively). No other correlations of BDNF and proBDNF levels with clinical and demographical variables have been detected. Discussion: Our results did not show any differences in BDNF and proBDNF levels between depression, mania, euthymia, and controls. Imbalance in BDNF/proBDNF signalling may be involved in pathogenesis of mood disorders. Further studies on larger groups are recommended. Grant was founded by National Science Center in Poland no 2011/03/D/NZ5/06146.

Keywords: bipolar disorder, Brain-Derived Neurotrophic Factor (BDNF), proBDNF, unipolar depression

Procedia PDF Downloads 216

Authors: Ashleigh Williams

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Micro- and nanoplastics’ (MNLPs) omnipresence and ecological accumulation is evident when surveying recent environmental impact studies. For example, in 2014 it was estimated that at least 52.3 trillion plastic microparticles are floating at sea, and scientists have even found plastics present remote Arctic ice and snow (5,6). Plastics have even found their way into precipitation, with more than 1000 tons of microplastic rain precipitating onto the Western United States in 2020. Even more recent studies evaluating the chemical safety of reusable plastic bottles found that hundreds of chemicals leached into the control liquid in the bottle (ddH2O, ph = 7) during a 24-hour time period. A consequence of the increased abundance in plastic waste in the air, land, and water every year is the bioaccumulation of MNLPs in ecosystems and trophic niches of the animal food chain, which could potentially cause increased direct and indirect exposure of humans to MNLPs via inhalation, ingestion, and dermal contact. Though the detrimental, toxic effects of MNLPs have been established in marine biota, much less is known about the potentially hazardous health effects of chronic MNLP ingestion in humans. Recent data indicate that long-term exposure to MNLPs could cause possible inflammatory and dysbiotic effects. However, toxicity seems to be largely dose-, as well as size-dependent. In addition, the transcytotic uptake of MNLPs through the intestinal epithelia in humans remain relatively unknown. To this point, the goal of the current study was to investigate the mechanisms of micro- and nanoplastic uptake and transcytosis of Polystyrene (PE) in human stem-cell derived, physiologically relevant in vitro intestinal model systems, and to compare the relative effect of particle size (30 nm, 100 nm, 500 nm and 1 µm), and concentration (0 µg/mL, 250 µg/mL, 500 µg/mL, 1000 µg/mL) on polystyrene MNLP uptake, transcytosis and intestinal epithelial model integrity. Observational and quantitative data obtained from confocal microscopy, immunostaining, transepithelial electrical resistance (TEER) measurements, cryosectioning, and ELISA cytokine assays of the proinflammatory cytokines Interleukin-6 and Interleukin-8 were used to evaluate the localization and transcytosis of polystyrene MNPs and its impact on epithelial integrity in human-derived intestinal in vitro model systems. The effect of Microfold (M) cell induction on polystyrene micro- and nanoparticle (MNP) uptake, transcytosis, and potential inflammation was also assessed and compared to samples grown under standard conditions. Microfold (M) cells, link the human intestinal system to the immune system and are the primary cells in the epithelium responsible for sampling and transporting foreign matter of interest from the lumen of the gut to underlying immune cells. Given the uptake capabilities of Microfold cells to interact both specifically and nonspecific to abiotic and biotic materials, it was expected that M- cell induced in vitro samples would have increased binding, localization, and potentially transcytosis of Polystyrene MNLPs across the epithelial barrier. Experimental results of this study would not only help in the evaluation of the plastic toxicity, but would allow for more detailed modeling of gut inflammation and the intestinal immune system.

Keywords: nanoplastics, enteroids, intestinal barrier, tissue engineering, microfold (M) cells

Procedia PDF Downloads 61

Authors: Ariunaa Sampilvanjil, Javzandulam Enkhbat, Chimegsaikhan Sodnompil, Oyungerel Sodjamts, Khulan Batbayasgalan, Altantsetseg Battulga, Tumenbayar Bataa

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Introduction: Prenatal stress has been linked to heightened allergy sensitivity in offspring. However, there is a notable absence of research on the mesovarium structure of offspring born from mothers subjected to cold stress during pregnancy. Understanding the impact of maternal cold stress on the mesovarium structure could provide valuable insights into reproductive health outcomes in offspring. Objective: This study aims to investigate structural changes in the mesovarium of offspring born from cold-stress affected rats. Material and Methods: 20 female Westar rats weighing around 200g were chosen and evenly divided into four containers; then 2-3 male rats were introduced to each container. The Papanicolaou method was used to estimate the spermatozoa and estrus period from vaginal swabs taken from female rats at 8:00 a.m. Female rats examined with presence of spermatozoa during the estrous phase of the estrous cycle are defined as pregnant. Pregnant rats are divided into experimental and control groups. The experimental group was stressed using the model of severe and chronic cold stress for 30 days. They were exposed to a cold stress for 3 hours each morning between 8:00 to 11:00 o’clock at a temperature of minus 15 degrees Celsius. The control group was kept under normal laboratory conditions. Newborn female rats from both experimental and control groups were selected. At 2 months of age, rats were euthanized by decapitation, and their mesovaria were collected. Tissues were fixed in 4% formalin, embedded in paraffin, and sectioned into 5μm thick slices. The sections were stained with H&E and digitized by digital microscope. Area of brown fat and inflammatory infiltrations were quantified using Image J software. The blood cortisol levels were measured using ELISA. Data are expressed as the mean ± standard error of the mean (SEM). The Mann-Whitney test was used to compare two groups. All analyses were performed using Prism (GraphPad Software). A p-value of < 0.05 was considered statistically significant. Result: Offspring born from stressed mothers exhibited significant physiological differences compared to the control group. Specifically: The body weight of offspring from stressed mothers was significantly lower than the control group (p=0.0002). Conversely, the cortisol level in offspring from stressed mothers was significantly higher (p=0.0446). Offspring born from stressed mothers showed a statistically significant increase in brown fat area compared to the control group (p=0.01). Additionally, offspring from stressed mothers had a significantly higher number of inflammatory infiltrates in their mesovarium compared to the control group (p<0.047). These results indicate the profound impact of maternal stress on offspring physiology, affecting body weight, stress hormone levels, metabolic characteristics, and inflammatory responses. Conclusion: Exposure to cold stress during pregnancy has significant repercussions on offspring physiology. Our findings demonstrate that cold stress exposure leads to increased blood cortisol levels, brown fat accumulation, and inflammatory cell infiltration in offspring. These results underscore the profound impact of maternal stress on offspring health and highlight the importance of mitigating environmental stressors during pregnancy to promote optimal offspring outcomes.

Keywords: brown fat, cold stress during pregnancy, inflammation, mesovarium

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Authors: Clara Tramuta, Lucia Decastelli, Elisa Barcucci, Sandra Fragassi, Samantha Lupi, Enrico Arletti, Melissa Bizzarri, Daniela Manila Bianchi

Abstract:

Introdution: Food allergies occurs, in the Western World, 2% of adults and up to 8% of children. The protection of allergic consumers is guaranted, in Eurrope, by Regulation (EU) No 1169/2011 of the European Parliament which governs the consumer's right to information and identifies 14 food allergens to be mandatory indicated on the label. Among these, mustard is a popular spice added to enhance the flavour and taste of foods. It is frequently present as an ingredient in spice blends, marinades, salad dressings, sausages, and other products. Hypersensitivity to mustard is a public health problem since the ingestion of even low amounts can trigger severe allergic reactions. In order to protect the allergic consumer, high performance methods are required for the detection of allergenic ingredients. Food safety laboratories rely on validated methods that detect hidden allergens in food to ensure the safety and health of allergic consumers. Here we present the test results for the validation and accreditation of a Real time PCR assay (RT-PCR: SPECIALfinder MC Mustard, Generon), for the detection of mustard traces in food. Materials and Methods. The method was tested on five classes of food matrices: bakery and pastry products (chocolate cookies), meats (ragù), ready-to-eat (mixed salad), dairy products (yogurt), grains, and milling products (rice and barley flour). Blank samples were spiked starting with the mustard samples (Sinapis Alba), lyophilized and stored at -18 °C, at a concentration of 1000 ppm. Serial dilutions were then prepared to a final concentration of 0.5 ppm, using the DNA extracted by ION Force FAST (Generon) from the blank samples. The Real Time PCR reaction was performed by RT-PCR SPECIALfinder MC Mustard (Generon), using CFX96 System (BioRad). Results. Real Time PCR showed a limit of detection (LOD) of 0.5 ppm in grains and milling products, ready-to-eat, meats, bakery, pastry products, and dairy products (range Ct 25-34). To determine the exclusivity parameter of the method, the ragù matrix was contaminated with Prunus dulcis (almonds), peanut (Arachis hypogaea), Glycine max (soy), Apium graveolens (celery), Allium cepa (onion), Pisum sativum (peas), Daucus carota (carrots), and Theobroma cacao (cocoa) and no cross-reactions were observed. Discussion. In terms of sensitivity, the Real Time PCR confirmed, even in complex matrix, a LOD of 0.5 ppm in five classes of food matrices tested; these values are compatible with the current regulatory situation that does not consider, at international level, to establish a quantitative criterion for the allergen considered in this study. The Real Time PCR SPECIALfinder kit for the detection of mustard proved to be easy to use and particularly appreciated for the rapid response times considering that the amplification and detection phase has a duration of less than 50 minutes. Method accuracy was rated satisfactory for sensitivity (100%) and specificity (100%) and was fully validated and accreditated. It was found adequate for the needs of the laboratory as it met the purpose for which it was applied. This study was funded in part within a project of the Italian Ministry of Health (IZS PLV 02/19 RC).

Keywords: allergens, food, mustard, real time PCR

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Authors: Elisa Coraggio, Dawei Han, Weiru Liu, Theo Tryfonas

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Water is one of the most important resources for human society. The world is currently undergoing a wave of urban growth, and pollution problems are of a great impact. Monitoring water quality is a key task for the future of the environment and human species. In recent times, researchers, using Smart Cities technologies are trying to mitigate the problems generated by the population growth in urban areas. The availability of huge amounts of data collected by a pervasive urban IoT can increase the transparency of decision making. Several services have already been implemented in Smart Cities, but more and more services will be involved in the future. Water quality monitoring can successfully be implemented in the urban IoT. The combination of water quality sensors, cloud computing, smart city infrastructure, and IoT technology can lead to a bright future for environmental monitoring. In the past decades, lots of effort has been put on monitoring and predicting water quality using traditional approaches based on manual collection and laboratory-based analysis, which are slow and laborious. The present study proposes a methodology for implementing a water quality prediction model using artificial intelligence techniques and comparing the results obtained with different algorithms. Furthermore, a 3D numerical model will be created using the software D-Water Quality, and simulation results will be used as a training dataset for the artificial intelligence algorithm. This study derives the methodology and demonstrates its implementation based on information and data collected at the floating harbour in the city of Bristol (UK). The city of Bristol is blessed with the Bristol-Is-Open infrastructure that includes Wi-Fi network and virtual machines. It was also named the UK ’s smartest city in 2017.In recent times, researchers, using Smart Cities technologies are trying to mitigate the problems generated by the population growth in urban areas. The availability of huge amounts of data collected by a pervasive urban IoT can increase the transparency of decision making. Several services have already been implemented in Smart Cities, but more and more services will be involved in the future. Water quality monitoring can successfully be implemented in the urban IoT. The combination of water quality sensors, cloud computing, smart city infrastructure, and IoT technology can lead to a bright future for the environment monitoring. In the past decades, lots of effort has been put on monitoring and predicting water quality using traditional approaches based on manual collection and laboratory-based analysis, which are slow and laborious. The present study proposes a new methodology for implementing a water quality prediction model using artificial intelligence techniques and comparing the results obtained with different algorithms. Furthermore, a 3D numerical model will be created using the software D-Water Quality, and simulation results will be used as a training dataset for the Artificial Intelligence algorithm. This study derives the methodology and demonstrate its implementation based on information and data collected at the floating harbour in the city of Bristol (UK). The city of Bristol is blessed with the Bristol-Is-Open infrastructure that includes Wi-Fi network and virtual machines. It was also named the UK ’s smartest city in 2017.

Keywords: artificial intelligence, hydroinformatics, numerical modelling, smart cities, water quality

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Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

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Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

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Authors: Federica Verna, Alessandra Pautasso, Maria Caramelli, Cristiana Maurella, Walter Mignone, Cristina Casalone

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Mosquito-Borne Diseases (MBDs) are dangerously increasing in prevalence, geographical distribution and severity, representing an emerging threat for both humans and animals. Interaction between multiple disciplines is needed for an effective early warning, surveillance and control of MBDs, according to the One Health concept. This work reports the integrated surveillance system enforced by IZSPLV in Piedmont, Liguria and Valle d’Aosta regions (Northwestern Italy) in order to control MDBs spread. Veterinary services and local human health authority are involved in an information network, to connect the surveillance of human clinical cases with entomological surveillance and veterinary monitoring in order to implement control measures in case of outbreak. A systematic entomological surveillance is carried out during the vector season using mosquitoes traps located in sites selected according to risk factors. Collected mosquitoes are counted, identified to species level by morphological standard classification keys and pooled by collection site, date and species with a maximum of 100 individuals. Pools are analyzed, after RNA extraction, by Real Time RT-PCR distinctive for West Nile Virus (WNV) Lineage 1 and Lineage 2, Real Time RT-PCR USUTU virus (USUV) and a traditional flavivirus End-point RT-PCR. Positive pools are sequenced and the related sequences employed to perform a basic local alignment search tool (BLAST) in the GenBank library. Positive samples are sent to the National Reference Centre for Animal Exotic Diseases (CESME, Teramo) for confirmation. With particular reference to WNV, after the confirmation, as provided by national legislation, control measures involving both local veterinary and human health services are activated: equine sera are randomly sampled within a 4 km radius from the positive collection sites and tested with ELISA kit and WNV NAT screening of blood donors is introduced. This surveillance network allowed to detect since 2011 USUV circulation in this area of Italy. WNV was detected in Piedmont and Liguria for the first time in 2014 in mosquitoes. During the 2015 vector season, we observed the expansion of its activity in Piedmont. The virus was detected in almost all Provinces both in mosquitoes (6 pools) and animals (19 equine sera, 4 birds). No blood bag tested resulted infected. The first neuroinvasive human case occurred too. Competent authorities should be aware of a potentially increased risk of MBDs activity during the 2016 vector season. This work shows that this surveillance network allowed to early detect the presence of MBDs in humans and animals, and provided useful information to public authorities, in order to apply control measures. Finally, an additional value of our diagnostic protocol is the ability to detect all viruses belonging to the Flaviviridae family, considering the emergence caused by other Flaviviruses in humans such as the recent Zika virus infection in South America. Italy has climatic and environmental features conducive to Zika virus transmission, the competent vector and many travellers from Brazil reported every year.

Keywords: integrated surveillance, mosquito borne disease, West Nile virus, Zika virus

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