Search results for: cellular communications

Authors: Lijuan Li

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Nitric oxide (NO) has become a fascinating entity in biological chemistry over the past few years. It is a gaseous lipophilic radical molecule that plays important roles in several physiological and pathophysiological processes in mammals, including activating the immune response, serving as a neurotransmitter, regulating the cardiovascular system, and acting as an endothelium-derived relaxing factor. NO functions in eukaryotes both as a signal molecule at nanomolar concentrations and as a cytotoxic agent at micromolar concentrations. The latter arises from the ability of NO to react readily with a variety of cellular targets leading to thiol S-nitrosation, amino acid N-nitrosation, and nitrosative DNA damage. Nitric oxide can readily bind to metals to give metal-nitrosyl (M-NO) complexes. Some of these species are known to play roles in biological NO storage and transport. These complexes have different biological, photochemical, or spectroscopic properties due to distinctive structural features. These recent discoveries have spawned a great interest in the development of transition metal complexes containing NO, particularly its iron complexes that are central to the role of nitric oxide in the body. Spectroscopic evidence would appear to implicate species of “Fe(NO)2+” type in a variety of processes ranging from polymerization, carcinogenesis, to nitric oxide stores. Our research focuses on isolation and structural studies of non-heme iron nitrosyls that mimic biologically active compounds and can potentially be used for anticancer drug therapy. We have shown that reactions between Fe(NO)2(CO)2 and a series of imidazoles generated new non-heme iron nitrosyls of the form Fe(NO)2(L)2 [L = imidazole, 1-methylimidazole, 4-methylimidazole, benzimidazole, 5,6-dimethylbenzimidazole, and L-histidine] and a tetrameric cluster of [Fe(NO)2(L)]4 (L=Im, 4-MeIm, BzIm, and Me2BzIm), resulted from the interactions of Fe(NO)2 with a series of substituted imidazoles was prepared. Recently, a series of sulfur bridged iron di nitrosyl complexes with the general formula of [Fe(µ-RS)(NO)2]2 (R = n-Pr, t-Bu, 6-methyl-2-pyridyl, and 4,6-dimethyl-2-pyrimidyl), were synthesized by the reaction of Fe(NO)2(CO)2 with thiols or thiolates. Their structures and properties were studied by IR, UV-vis, 1H-NMR, EPR, electrochemistry, X-ray diffraction analysis and DFT calculations. IR spectra of these complexes display one weak and two strong NO stretching frequencies (νNO) in solution, but only two strong νNO in solid. DFT calculations suggest that two spatial isomers of these complexes bear 3 Kcal energy difference in solution. The paramagnetic complexes [Fe2(µ-RS)2(NO)4]-, have also been investigated by EPR spectroscopy. Interestingly, the EPR spectra of complexes exhibit an isotropic signal of g = 1.998 - 2.004 without hyperfine splitting. The observations are consistent with the results of calculations, which reveal that the unpaired electron dominantly delocalize over the two sulfur and two iron atoms. The difference of the g values between the reduced form of iron-sulfur clusters and the typical monomeric di nitrosyl iron complexes is explained, for the first time, by of the difference in unpaired electron distributions between the two types of complexes, which provides the theoretical basis for the use of g value as a spectroscopic tool to differentiate these biologically active complexes.

Keywords: di nitrosyl iron complex, metal nitrosyl, non-heme iron, nitric oxide

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Authors: Siddhant Sethi, Yasuharu Takashima, Shigetaka Nakamura, Kenzo Fujimoto

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Site-directed mutagenesis is a renowned technique to introduce specific mutations in the genome. To achieve site-directed mutagenesis, many chemical and enzymatic approaches have been reported in the past like disulphite induced genome editing, CRISPR-Cas9, TALEN etc. The chemical methods are invasive whereas the enzymatic approaches are time-consuming and expensive. Most of these techniques are unusable in the cellular application due to their toxicity and other limitations. Photo-chemical cytosine deamination, introduced in 2010, is one of the major technique for enzyme-free single-point mutation of cytosine to uracil in DNA and RNA, wherein, 3-cyanovinylcarbazole nucleoside (CNVK) containing oligodeoxyribonucleotide (ODN) having CNVK at -1 position to that of target cytosine is reversibly crosslinked to target DNA strand using 366 nm and then incubated at 90ºC to accommodate deamination. This technique is superior to enzymatic methods of site-directed mutagenesis but has a disadvantage that it requires the use of high temperature for the deamination step which restricts its applicability in the in vivo applications. This study has been focused on improving the technique by reducing the temperature required for deamination. Firstly, the photo-cross-linker, CNVK has been modified by replacing cyano group attached to vinyl group with methyl ester (OMeVK), amide (NH2VK), and carboxylic acid (OHVK) to observe the acceleration in the deamination of target cytosine cross-linked to vinylcarbazole derivative. Among the derivatives, OHVK has shown 2 times acceleration in deamination reaction as compared to CNVK, while the other two derivatives have shown deceleration towards deamination reaction. The trend of rate of deamination reaction follows the same order as that of hydrophilicity of the vinylcarbazole derivatives. OHVK being most hydrophilic has shown highest acceleration while OMeVK is least hydrophilic has proven to be least active for deamination. Secondly, in the related study, the counter-base of the target cytosine, guanine has been replaced by inosine, 2-aminopurine, nebularine, and 5-nitroindole having distinct hydrogen bonding patterns with target cytosine. Among the ODNs with these counter bases, ODN with inosine has shown 12 fold acceleration towards deamination of cytosine cross-linked to CNVK at physiological conditions as compared to guanosine. Whereas, when 2-aminopurine, nebularine, and 5-nitroindole were used, no deamination reaction took place. It can be concluded that inosine has potential to be used as the counter base of target cytosine for the CNVK mediated photo-cross-linking induced deamination of cytosine. The increase in rate of deamination reaction has been attributed to pattern and number of hydrogen bonding between the cytosine and counter base. One of the important factor is presence of hydrogen bond between exo-cyclic amino group of cytosine and the counter base. These results will be useful for development of more efficient technique for site-directed mutagenesis for C → U transformations in the DNA/RNA which might be used in the living system for treatment of various genetic disorders and genome engineering for making designer and non-native proteins.

Keywords: C to U transformation, DNA editing, genome engineering, ultra-fast photo-cross-linking

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Authors: Fatemeh Asadi Farsad, Sima Modirkhameneh

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The purpose of this study was to explore the pattern of Psychological Well-Being (PWB) of Iranian male and female EFL teachers during the pandemic. It was intended to see if such a drastic change in the context and mode of teaching affects teachers' PWB. Furthermore, the possible difference between the six elements of PWB of Iranian EFL male vs. female teachers during the pandemic was investigated. The other purpose was to find out the EFL teachers’ perceptions of any modifications, and factors leading to such modifications in their PWB during pandemic. For the purpose of this investigation, a total of 81 EFL teachers (59 female, 22 male) with an age range of 25 to 35 were conveniently sampled from different cities in Iran. Ryff’s PWB questionnaire was sent to participant teachers through online platforms to elicit data on their PWB. As for their perceptions on the possible modifications and the factors involved in PWB during pandemic, a set of semi-structured interviews were run among both sample groups. The findings revealed that male EFL teachers had the highest mean on personal growth, followed by purpose of life, and self-acceptance and the lowest mean on environmental mastery. With a slightly similar pattern, female EFL teachers had the highest mean on personal growth, followed by purpose in life, and positive relationship with others with the lowest mean on environmental mastery. However, no significant difference was observed between the male and female groups’ overall means on elements of PWB. Additionally, participants perceived that their anxiety level in online classes altered due to factors like (1) Computer literacy skills, (2) Lack of social communications and interactions with colleagues and students, (3) Online class management, (4) Overwhelming workloads, and (5) Time management. The study ends with further suggestions as regards effective online teaching preparation considering teachers PWB, especially at severe situations such as covid-19 pandemic. The findings offer to determine the reformations of educational policies concerning enhancing EFL teachers’ PWB through computer literacy courses and stress management courses. It is also suggested that to proactively support teachers’ mental health, it is necessary to provide them with advisors and psychologists if possible for free. Limitations: One limitation is the small number of participants (81), suggesting that future replications should include more participants for reliable findings. Another limitation is the gender imbalance, which future studies should address to yield better outcomes. Furthermore, Limited data gathering tools suggest using observations, diaries, and narratives for more insights in future studies. The study focused on one model of PWB, calling for further research on other models in the literature. Considering the wide effect of the COVID-19 pandemic, future studies should consider additional variables (e.g., teaching experience, age, income) to understand Iranian EFL teachers’ vulnerabilities and strengths better.

Keywords: online teaching, psychological well-being, female and male EFL teachers, pandemic

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Authors: Iryna Atamaniuk, Hannah Boysen, Nils Wieczorek, Natalia Politaeva, Iuliia Bazarnova, Kerstin Kuchta

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Over the last decade due to climate change and a strategy of natural resources preservation, the interest for the aquatic biomass has dramatically increased. Along with mitigation of the environmental pressure and connection of waste streams (including CO₂ and heat emissions), microalgae bioeconomy can supply food, feed, as well as the pharmaceutical and power industry with number of value-added products. Furthermore, in comparison to conventional biomass, microalgae can be cultivated in wide range of conditions without compromising food and feed production, thus addressing issues associated with negative social and the environmental impacts. This paper presents the state-of-the art technology for microalgae bioeconomy from cultivation process to production of valuable components and by-streams. Microalgae Chlorella sorokiniana were cultivated in the pilot-scale innovation concept in Hamburg (Germany) using different systems such as race way pond (5000 L) and flat panel reactors (8 x 180 L). In order to achieve the optimum growth conditions along with suitable cellular composition for the further extraction of the value-added components, process parameters such as light intensity, temperature and pH are continuously being monitored. On the other hand, metabolic needs in nutrients were provided by addition of micro- and macro-nutrients into a medium to ensure autotrophic growth conditions of microalgae. The cultivation was further followed by downstream process and extraction of lipids, proteins and saccharides. Lipids extraction is conducted in repeated-batch semi-automatic mode using hot extraction method according to Randall. As solvents hexane and ethanol are used at different ratio of 9:1 and 1:9, respectively. Depending on cell disruption method along with solvents ratio, the total lipids content showed significant variations between 8.1% and 13.9 %. The highest percentage of extracted biomass was reached with a sample pretreated with microwave digestion using 90% of hexane and 10% of ethanol as solvents. Proteins content in microalgae was determined by two different methods, namely: Total Kejadahl Nitrogen (TKN), which further was converted to protein content, as well as Bradford method using Brilliant Blue G-250 dye. Obtained results, showed a good correlation between both methods with protein content being in the range of 39.8–47.1%. Characterization of neutral and acid saccharides from microalgae was conducted by phenol-sulfuric acid method at two wavelengths of 480 nm and 490 nm. The average concentration of neutral and acid saccharides under the optimal cultivation conditions was 19.5% and 26.1%, respectively. Subsequently, biomass residues are used as substrate for anaerobic digestion on the laboratory-scale. The methane concentration, which was measured on the daily bases, showed some variations for different samples after extraction steps but was in the range between 48% and 55%. CO₂ which is formed during the fermentation process and after the combustion in the Combined Heat and Power unit can potentially be used within the cultivation process as a carbon source for the photoautotrophic synthesis of biomass.

Keywords: bioeconomy, lipids, microalgae, proteins, saccharides

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Authors: Jessica M. Black

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Although much of natural and social science research aims to enhance human flourishing and address social problems, the training within the two fields is significantly different across theory, methodology, and implementation of results. Social scientists are trained in social, psychological, and to the extent that it is relevant to their discipline, spiritual development, theory, and accompanying methodologies. They tend not to receive training or learn about accompanying methodology related to interrogating human development and social problems from a biological perspective. On the other hand, those in the natural sciences, and for the purpose of this work, human biological sciences specifically – biology, neuroscience, genetics, epigenetics, and physiology – are often trained first to consider cellular development and related methodologies, and may not have opportunity to receive formal training in many of the foundational principles that guide human development, such as systems theory or person-in-environment framework, methodology related to tapping both proximal and distal psycho-social-spiritual influences on human development, and foundational principles of equity, justice and inclusion in research design. There is a need for disciplines heretofore siloed to know one another, to receive streamlined, easy to access training in theory and methods from one another and to learn how to build interdisciplinary teams that can speak and act upon a shared research language. Team science is more essential than ever, as are transdisciplinary approaches to training and research design. This study explores the use of a methodological toolbox that natural and social scientists can use by employing a decision-making tree regarding project aims, costs, and participants, among other important study variables. The decision tree begins with a decision about whether the researcher wants to learn more about social sciences approaches or biological approaches to study design. The toolbox and platform are flexible, such that users could also choose among modules, for instance, reviewing epigenetics or community-based participatory research even if those are aspects already a part of their home field. To start, both natural and social scientists would receive training on systems science, team science, transdisciplinary approaches, and translational science. Next, social scientists would receive training on grounding biological theory and the following methodological approaches and tools: physiology, (epi)genetics, non-invasive neuroimaging, invasive neuroimaging, endocrinology, and the gut-brain connection. Natural scientists would receive training on grounding social science theory, and measurement including variables, assessment and surveys on human development as related to the developing person (e.g., temperament and identity), microsystems (e.g., systems that directly interact with the person such as family and peers), mesosystems (e.g., systems that interact with one another but do not directly interact with the individual person, such as parent and teacher relationships with one another), exosystems (e.g., spaces and settings that may come back to affect the individual person, such as a parent’s work environment, but within which the individual does not directly interact, macrosystems (e.g., wider culture and policy), and the chronosystem (e.g., historical time, such as the generational impact of trauma). Participants will be able to engage with the toolbox and one another to foster increased transdisciplinary work

Keywords: methodology, natural science, social science, transdisciplinary

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Authors: Anna Höfer, Johannes Herrmann, Patrick Meybohm, Christopher Lotz

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Mitochondria are pivotal for energy supply and regulation of cellular functions. Deficiencies of mitochondrial metabolism have been implicated in diverse stressful conditions including infections. Platelets are key mediators for thrombo-inflammation during development and resolution of acute respiratory distress syndrome (ARDS). Previous data point to an exhausted platelet phenotype in critically-ill patients with coronavirus 19 disease (COVID-19) impacting the course of disease. The objective of this work was to characterize platelet mitochondrial metabolism in patients suffering from COVID-19 ARDSA longitudinal analysis of platelet mitochondrial metabolism in 24 patients with COVID-19 induced ARDS compared to 35 healthy controls (ctrl) was performed. Blood samples were analyzed at two time points (t1=day 1; t2=day 5-7 after study inclusion). The activity of mitochondrial citrate synthase was photometrically measured. The impact of oxidative stress on mitochondrial permeability was assessed by a photometric calcium-induced swelling assay and the activity of superoxide dismutase (SOD) by a SOD assay kit. The amount of protein carbonylation and the activity of mitochondria complexes I-IV were photometrically determined. Levels of interleukins (IL)-1α, IL-1β and tumor necrosis factor (TNF-) α were measured by a Multiplex assay kit. Median age was 54 years, 63 % were male and BMI was 29.8 kg/m2. SOFA (12; IQR: 10-15) and APACHE II (27; IQR: 24-30) indicated critical illness. Median Murray Score was 3.4 (IQR: 2.8-3.4), 21/24 (88%) required mechanical ventilation and V-V ECMO support in 14/24 (58%). Platelet counts in ARDS did not change during ICU stay (t1: 212 vs. t2: 209 x109/L). However, mean platelet volume (MPV) significantly increased (t1: 10.6 vs. t2: 11.9 fL; p<0.0001). Citrate synthase activity showed no significant differences between ctrl and ARDS patients. Calcium induced swelling was more pronounced in patients at t1 compared to t2 and to ctrl (50µM; t1: 0.006 vs. ctrl: 0.016 ΔOD; p=0.001). The amount of protein carbonylation as marker for irreversible proteomic modification constantly increased during ICU stay and compared to ctrl., without reaching significance. In parallel, superoxid dismutase activity gradually declined during ICU treatment vs. ctrl (t2: - 29 vs. ctrl.: - 17 %; p=0.0464). Complex I analysis revealed significantly stronger activity in ARDS vs. ctrl. (t1: 0.633 vs. ctrl.: 0.415 ΔOD; p=0.0086). There were no significant differences in complex II, III or IV activity in platelets from ARDS patients compared to ctrl. IL-18 constantly increased during the observation period without reaching significance. IL-1α and TNF-α did not differ from ctrl. However, IL-1β levels were significantly elevated in ARDS (t1: 16.8; t2: 16.6 vs. ctrl.: 12.4 pg/mL; p1=0.0335, p2=0.0032). This study reveals new insights in platelet mitochondrial metabolism during COVID-19 caused ARDS. it data point towards enhanced platelet activity with a pronounced turnover rate. We found increased activity of mitochondria complex I and evidence for enhanced oxidative stress. In parallel, protective mechanisms against oxidative stress were narrowed with elevated levels of IL-1β likely causing a pro-apoptotic environment. These mechanisms may contribute to platelet exhaustion in ARDS.

Keywords: acute respiratory distress syndrome (ARDS), coronavirus 19 disease (COVID-19), oxidative stress, platelet mitochondrial metabolism

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Authors: Marjorie Miguel

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Globalization offers better career opportunities hence demands more competent professionals efficient for the job. With the transformation of the world industry from competition to collaboration coupled with the rapid development in the field of science and technology, engineers need not only to be technically proficient, but also multilingual-skilled: two characteristics that a global engineer possesses. English often serves as the global language between people from different cultures being the medium mostly used in international business. Ironically, most universities worldwide adapt engineering curriculum heavily built around the language of mathematics not realizing that the goal of an engineer is not only to create and design, but more importantly to promote his creations and designs to the general public through effective communication. This premise led to some developments in the teaching process of English subjects in the tertiary level which include the integration of the technical knowledge related to the area of specialization of the students in the English subjects that they are taking. This is also known as English for Specific Purposes. This study focused on the development of English for Specific Purposes-Based Instructional Materials for Engineering Students of Bulacan State University (BulSU). The materials were tailor-made in which the contents and structure were designed to meet the specific needs of the students as well as the industry. Based on the needs analysis, the needs of the students and the industry were determined to make the study descriptive in nature. The major respondents included fifty engineering students and ten professional engineers from selected institutions. The needs analysis was done and the results showed the common writing difficulties of the students and the writing skills needed among the engineers in the industry. The topics in the instructional materials were established after the needs analysis was conducted. Simple statistical treatment including frequency distribution, percentages, mean, standard deviation, and weighted mean were used. The findings showed that the greatest number of the respondents had an average proficiency rating in writing, and the much-needed skills that must be developed by the engineers are directly related to the preparation and presentation of technical reports about their projects, as well as to the different communications they transmit to their colleagues and superiors. The researcher undertook the following phases in the development of the instructional materials: a design phase, development phase, and evaluation phase. Evaluations are given by some college instructors about the instructional materials generally helped in its usefulness and significance making the study beneficial not only as a career enhancer for BulSU engineering students, but also creating the university one of the educational institutions ready for the new millennium.

Keywords: English for specific purposes, instructional materials, needs analysis, write (right) job description

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Authors: Diamant Irene, Shklarnik Batya

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In on to its widespread effects on health and economic issues, Covid-19 shook the work and employment world. Among the prominent changes during the pandemic is the work-from-home trend, complete or partial, as part of social distancing. In fact, these changes accelerated an existing tendency of work flexibility already underway before the pandemic. Technology and means of advanced communications led to a re-assessment of “place of work” as a physical space in which work takes place. Today workers can remotely carry out meetings, manage projects, work in groups, and different research studies point to the fact that this type of work has no adverse effect on productivity. However, from the worker’s perspective, despite numerous advantages associated with work from home, such as convenience, flexibility, and autonomy, various drawbacks have been identified such as loneliness, reduction of commitment, home-work boundary erosion, all risk factors relating to the quality of life and burnout. Thus, a real need has arisen in exploring differences in work-from-home experiences and understanding the relationship between psychological characteristics and the prevalence of burnout. This understanding may be of significant value to organizations considering a future hybrid work model combining in-office and remote working. Based on Hobfoll’s Theory of Conservation of Resources, we hypothesized that burnout would mainly be found among workers whose physical remoteness from the workplace threatens or hinders their ability to retain significant individual resources. In the present study, we compared fully remote and partially remote workers (hybrid work), and we examined psychological characteristics and their connection to the formation of burnout. Based on the conceptualization of Place Attachment as the cognitive-emotional bond of an individual to a meaningful place and the need to maintain closeness to it, we assumed that individuals characterized with Place Attachment to the workplace would suffer more from burnout when working from home. We also assumed that extrovert individuals, characterized by the need of social interaction at the workplace and individuals with segmentationpreference – a need for separation between different life domains, would suffer more from burnout, especially among fully remote workers relative to partially remote workers. 194 workers, of which 111 worked from home in full and 83 worked partially from home, aged 19-53, from different sectors, were tested using an online questionnaire through social media. The results of the study supported our assumptions. The repercussions of these findings are discussed, relating to future occupational experience, with an emphasis on suitable occupational adjustment according to the psychological characteristics and needs of workers.

Keywords: working from home, burnout, place attachment, extraversion, segmentation preference, Covid-19

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Authors: Marcin Dębowski, Marcin Zieliński, Mirosław Krzemieniewski

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As methane fermentation is a complex series of successive biochemical transformations, its subsequent stages are determined, to a various extent, by physical and chemical factors. A specific state of equilibrium is being settled in the functioning fermentation system between environmental conditions and the rate of biochemical reactions and products of successive transformations. In the case of physical factors that influence the effectiveness of methane fermentation transformations, the key significance is ascribed to temperature and intensity of biomass agitation. Among the chemical factors, significant are pH value, type, and availability of the culture medium (to put it simply: the C/N ratio) as well as the presence of toxic substances. One of the important elements which influence the effectiveness of methane fermentation is the pre-treatment of organic substrates and the mode in which the organic matter is made available to anaerobes. Out of all known and described methods for organic substrate pre-treatment before methane fermentation process, the ultrasound disintegration is one of the most interesting technologies. Investigations undertaken on the ultrasound field and the use of installations operating on the existing systems result principally from very wide and universal technological possibilities offered by the sonication process. This physical factor may induce deep physicochemical changes in ultrasonicated substrates that are highly beneficial from the viewpoint of methane fermentation processes. In this case, special role is ascribed to disintegration of biomass that is further subjected to methane fermentation. Once cell walls are damaged, cytoplasm and cellular enzymes are released. The released substances – either in dissolved or colloidal form – are immediately available to anaerobic bacteria for biodegradation. To ensure the maximal release of organic matter from dead biomass cells, disintegration processes are aimed to achieve particle size below 50 μm. It has been demonstrated in many research works and in systems operating in the technical scale that immediately after substrate supersonication the content of organic matter (characterized by COD, BOD5 and TOC indices) was increasing in the dissolved phase of sedimentation water. This phenomenon points to the immediate sonolysis of solid substances contained in the biomass and to the release of cell material, and consequently to the intensification of the hydrolytic phase of fermentation. It results in a significant reduction of fermentation time and increased effectiveness of production of gaseous metabolites of anaerobic bacteria. Because disintegration of Virginia fanpetals biomass via ultrasounds applied in order to intensify its conversion is a novel technique, it is often underestimated by exploiters of agri-biogas works. It has, however, many advantages that have a direct impact on its technological and economical superiority over thus far applied methods of biomass conversion. As for now, ultrasound disintegrators for biomass conversion are not produced on the mass-scale, but by specialized groups in scientific or R&D centers. Therefore, their quality and effectiveness are to a large extent determined by their manufacturers’ knowledge and skills in the fields of acoustics and electronic engineering.

Keywords: ultrasound disintegration, biomass, methane fermentation, biogas, Virginia fanpetals

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Authors: Marcelo T. Okano, Oduvaldo Vendrametto, Osmildo S. Santos, Marcelo E. Fernandes, Heide Landi

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Teaching of entrepreneurship and innovation in Brazilian universities has increased in recent years due to several factors such as the emergence of disciplines like biotechnology increased globalization reduced basic funding and new perspectives on the role of the university in the system of knowledge production Innovation is increasingly seen as an evolutionary process that involves different institutional spheres or sectors in society Entrepreneurship is a milestone on the road towards economic progress, and makes a huge contribution towards the quality and future hopes of a sector, economy or even a country. Entrepreneurship is as important in small and medium-sized enterprises (SMEs) and local markets as in large companies, and national and international markets, and is just as key a consideration for public companies as or private organizations. Entrepreneurship helps to encourage the competition in the current environment that leads to the effects of globalization. There is an increasing tendency for government policy to promote entrepreneurship for its apparent economic benefit. Accordingly, governments seek to employ entrepreneurship education as a means to stimulate increased levels of economic activity. Entrepreneurship education and training (EET) is growing rapidly in universities and colleges throughout the world, and governments are supporting it both directly and through funding major investments in advice-provision to would-be entrepreneurs and existing small businesses. The Triple Helix of university–industry–government relations is compared with alternative models for explaining the current research system in its social contexts. Communications and negotiations between institutional partners generate an overlay that increasingly reorganizes the underlying arrangements. To achieve the objective of this research was a survey of the literature on the entrepreneurship and innovation and then a field research with 100 students of Fatec. To collect the data needed for analysis, we used the exploratory research of a qualitative nature. We asked to respondents what degree of knowledge over ten related to entrepreneurship and innovation topics, responses were answered in a Likert scale with 4 levels, none, small, medium and large. We can conclude that the terms such as entrepreneurship and innovation are known by most students because the university propagates them across disciplines, lectures, and institutes innovation. The more specific items such as canvas and Design thinking model are unknown by most respondents. The importance of the University in teaching innovation and entrepreneurship in the transmission of this knowledge to the students in order to equalize the knowledge. As a future project, these items will be re-evaluated to create indicators for measuring the knowledge level.

Keywords: Brazilian universities, entrepreneurship, innovation, entrepreneurship, globalization

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Authors: G. M. Vlasceanu, H. Iovu, E. Vasile, M. Ionita

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Herein, the synthesis and characterization of chitosan-gelatin highly porous scaffold reinforced with graphene oxide, and hydroxyapatite (HAp), crosslinked with genipin was targeted. In tissue engineering, chitosan and gelatin are two of the most robust biopolymers with wide applicability due to intrinsic biocompatibility, biodegradability, low antigenicity properties, affordability, and ease of processing. HAp, per its exceptional activity in tuning cell-matrix interactions, is acknowledged for its capability of sustaining cellular proliferation by promoting bone-like native micro-media for cell adjustment. Genipin is regarded as a top class cross-linker, while graphene oxide (GO) is viewed as one of the most performant and versatile fillers. The composites with natural bone HAp/biopolymer ratio were obtained by cascading sonochemical treatments, followed by uncomplicated casting methods and by freeze-drying. Their structure was characterized by Fourier Transform Infrared Spectroscopy and X-ray Diffraction, while overall morphology was investigated by Scanning Electron Microscopy (SEM) and micro-Computer Tomography (µ-CT). Ensuing that, in vitro enzyme degradation was performed to detect the most promising compositions for the development of in vivo assays. Suitable GO dispersion was ascertained within the biopolymer mix as nanolayers specific signals lack in both FTIR and XRD spectra, and the specific spectral features of the polymers persisted with GO load enhancement. Overall, correlations between the GO induced material structuration, crystallinity variations, and chemical interaction of the compounds can be correlated with the physical features and bioactivity of each composite formulation. Moreover, the HAp distribution within follows an auspicious density gradient tuned for hybrid osseous/cartilage matter architectures, which were mirrored in the mice model tests. Hence, the synthesis route of a natural polymer blend/hydroxyapatite-graphene oxide composite material is anticipated to emerge as influential formulation in bone tissue engineering. Acknowledgement: This work was supported by the project 'Work-based learning systems using entrepreneurship grants for doctoral and post-doctoral students' (Sisteme de invatare bazate pe munca prin burse antreprenor pentru doctoranzi si postdoctoranzi) - SIMBA, SMIS code 124705 and by a grant of the National Authority for Scientific Research and Innovation, Operational Program Competitiveness Axis 1 - Section E, Program co-financed from European Regional Development Fund 'Investments for your future' under the project number 154/25.11.2016, P_37_221/2015. The nano-CT experiments were possible due to European Regional Development Fund through Competitiveness Operational Program 2014-2020, Priority axis 1, ID P_36_611, MySMIS code 107066, INOVABIOMED.

Keywords: biopolymer blend, ectopic osteoinduction, graphene oxide composite, hydroxyapatite

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Authors: Diego R. Boiarski, Camila M. Toigo, Thais M. Sobjak, Andrey F. P. Santos, Silvia Romao, Ana T. B. Guimaraes

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Anthropogenic activities promote changes in the community’s structures and decrease the species abundance of amphibians. Biological communities of fluvial systems are assemblies of organisms that have adapted to regional conditions, including the physical environment and food resources, and are further refined through interactions with other species. The aim of this study was to assess neurotoxic alterations and in the antioxidant system on tadpoles of Lithobates catesbeianus exposed to waters from Cascavel River, in the south of Brazil. A total of 420 L of water was collected from the Cascavel River, 140 L from each of the three different locations: Site 1 – headwater; Site 2 – stretch of the stream that runs through an urbanized area; Site 3 – a stretch from the rural area. Twelve tadpoles were acclimated in each aquarium (100 L of water) for seven days. The water from each aquarium was replaced with the ones sampled from the river, except the one from the control aquarium. After seven days, a portion of the liver was removed and conditioned for ChE, SOD, CAT and LPO analysis; other part of the tissue was conditioned for histological analysis. The statistical analysis performed was one-way ANOVA, followed by post-hoc Tukey-HSD test, and the multivariate principal components analysis. It was not observed any neurotoxic effect, but a slight increase in SOD activity and elevation of CAT activity in both urban and rural environment. A decrease in LPO reaction was detected, mainly among the tadpoles exposed to the waters from the rural area. The results of the present study demonstrate the alteration of the antioxidant system, as well as liver histopathologies in tadpoles exposed mainly to waters collected in urban and rural environments. These alterations may cause the reduction in the velocity of the metamorphosis process from the tadpoles. Further, were observed histological alterations, highlighting necrotic areas mainly among the animals exposed to urban waters. Those damages can lead to metabolic dysfunction, interfering with survival capacity, diminishing not only individual fitness but for the whole population. In the interpretation synthesis of all biomarkers, the cellular damage gradient is perceptible, characterized by the variables related to the antioxidant system, due to the flow direction of the stream. This result is indicative that along the course of the creek occurs dumping of organic material, which promoted an acute response upon tadpoles of L. catesbeianus. and it was also observed the difference in tissue damage between the experimental groups and the control group, the latter presenting histological alterations, but to a lesser degree than the animals exposed to the waters of the Cascavel river. These damages, caused by reactive oxygen species possibly resulting from the contamination by organic compounds, can lead the animals to a series of metabolic dysfunctions, interfering with its metamorphosis capacity. Interruption of metamorphosis may affect survival, which may impair its growth, development and reproduction, diminishing not only the fitness of each individual but in a long-term, to the entire population.

Keywords: American bullfrog, histopathology, oxidative stress, urban creeks pollution

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Authors: Iman Farasat, Howard M. Salis

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Engineered genetic circuits reprogram cellular behavior to act as living computers with applications in detecting cancer, creating self-controlling artificial tissues, and dynamically regulating metabolic pathways. Phenemenological models are often used to simulate and design genetic circuit behavior towards a desired behavior. While such models assume that each circuit component’s function is modular and independent, even small changes in a circuit (e.g. a new promoter, a change in transcription factor expression level, or even a new media) can have significant effects on the circuit’s function. Here, we use statistical thermodynamics to account for the several factors that control transcriptional regulation in bacteria, and experimentally demonstrate the model’s accuracy across 825 measurements in several genetic contexts and hosts. We then employ our first principles model to design, experimentally construct, and characterize a family of signal amplifying genetic circuits (genetic OpAmps) that expand the dynamic range of cell sensors. To develop these models, we needed a new approach to measuring the in vivo binding free energies of transcription factors (TFs), a key ingredient of statistical thermodynamic models of gene regulation. We developed a new high-throughput assay to measure RNA polymerase and TF binding free energies, requiring the construction and characterization of only a few constructs and data analysis (Figure 1A). We experimentally verified the assay on 6 TetR-homolog repressors and a CRISPR/dCas9 guide RNA. We found that our binding free energy measurements quantitatively explains why changing TF expression levels alters circuit function. Altogether, by combining these measurements with our biophysical model of translation (the RBS Calculator) as well as other measurements (Figure 1B), our model can account for changes in TF binding sites, TF expression levels, circuit copy number, host genome size, and host growth rate (Figure 1C). Model predictions correctly accounted for how these 8 factors control a promoter’s transcription rate (Figure 1D). Using the model, we developed a design framework for engineering multi-promoter genetic circuits that greatly reduces the number of degrees of freedom (8 factors per promoter) to a single dimensionless unit. We propose the Ptashne (Pt) number to encapsulate the 8 co-dependent factors that control transcriptional regulation into a single number. Therefore, a single number controls a promoter’s output rather than these 8 co-dependent factors, and designing a genetic circuit with N promoters requires specification of only N Pt numbers. We demonstrate how to design genetic circuits in Pt number space by constructing and characterizing 15 2-repressor OpAmp circuits that act as signal amplifiers when within an optimal Pt region. We experimentally show that OpAmp circuits using different TFs and TF expression levels will only amplify the dynamic range of input signals when their corresponding Pt numbers are within the optimal region. Thus, the use of the Pt number greatly simplifies the genetic circuit design, particularly important as circuits employ more TFs to perform increasingly complex functions.

Keywords: transcription factor, synthetic biology, genetic circuit, biophysical model, binding energy measurement

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Authors: Tesfaye Dagne, Dagmawit Solomon, Firmaye Bogale, Yosef Gebreyohannes, Samson Mideksa, Mamuye Hadis, Desalegn Ararso, Ermias Woldie, Tsegaye Getachew, Sabit Ababor, Zelalem Kebede

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Globally, road traffic accident (RTA) is causing millions of deaths and injuries every year. It is one of the leading causes of death among people of all age groups and the problem is worse among young reproductive age group. Moreover the problem is increasing with an increasing number of vehicles. The majority of the problem happen in low and middle income countries (LMIC), even if the number of vehicles in these countries is low compared to their population. So, the objective of this paper is to summarize the best available evidence on interventions that can reduce road traffic accidents in low and middle income countries (LMIC). Method: A rapid evidence synthesis approach adapted from the SURE Rapid Response Service was applied to search, appraise and summarize the best available evidence on effective intervention in reducing road traffic injury. To answer the question under review, we searched for relevant studies from databases including PubMed, the Cochrane Library, TRANSPORT, Health system evidence, Epistemonikos, and SUPPORT summary. The following key terms were used for searching: Road traffic accident, RTA, Injury, Reduc*, Prevent*, Minimiz*, “Low and middle-income country”, LMIC. We found 18 articles through a search of different databases mentioned above. After screening for the titles and abstracts of the articles, four of them which satisfy the inclusion criteria were included in the final review. Then we appraised and graded the methodological quality of systematic reviews that are deemed to be highly relevant using AMSTAR. Finding: The identified interventions to reduce road traffic accidents were legislation and enforcement, public awareness/education, speed control/ rumble strips, road improvement, mandatory motorcycle helmet, graduated driver license, street lighting. Legislation and Enforcement: Legislation focusing on mandatory motorcycle helmet usage, banning cellular phone usage when driving, seat belt laws, decreasing the legal blood alcohol content (BAC) level from 0.06 g/L to 0.02 g/L bring the best result where enforcement is there. Public Awareness/Education: focusing on seat belt use, child restraint use, educational training in health centers and schools/universities, and public awareness with media through the distribution of videos, posters/souvenirs, and pamphlets are effective in the short run. Speed Control: through traffic calming bumps, or speed bumps, rumbled strips are effective in reducing accidents and fatality. Mandatory Motorcycle Helmet: is associated with reduction in mortality. Graduated driver’s license (GDL): reduce road traffic injury by 19%. Street lighting: is a low-cost intervention which may reduce road traffic accidents.

Keywords: evidence synthesis, injury, rapid review, reducing, road traffic accident

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Authors: Marcin Zieliński, Marcin Dębowski, Mirosław Krzemieniewski

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The anaerobic degradation of substrates is limited especially by the rate and effectiveness of the first (hydrolytic) stage of fermentation. This stage may be intensified through pre-treatment of substrate aimed at disintegration of the solid phase and destruction of substrate tissues and cells. The most frequently applied criterion of disintegration outcomes evaluation is the increase in biogas recovery owing to the possibility of its use for energetic purposes and, simultaneously, recovery of input energy consumed for the pre-treatment of substrate before fermentation. Hydrodynamic cavitation is one of the methods for organic substrate disintegration that has a high implementation potential. Cavitation is explained as the phenomenon of the formation of discontinuity cavities filled with vapor or gas in a liquid induced by pressure drop to the critical value. It is induced by a varying field of pressures. A void needs to occur in the flow in which the pressure first drops to the value close to the pressure of saturated vapor and then increases. The process of cavitation conducted under controlled conditions was found to significantly improve the effectiveness of anaerobic conversion of organic substrates having various characteristics. This phenomenon allows effective damage and disintegration of cellular and tissue structures. Disintegration of structures and release of organic compounds to the dissolved phase has a direct effect on the intensification of biogas production in the process of anaerobic fermentation, on reduced dry matter content in the post-fermentation sludge as well as a high degree of its hygienization and its increased susceptibility to dehydration. A device the efficiency of which was confirmed both in laboratory conditions and in systems operating in the technical scale is a hydrodynamic generator of cavitation. Cavitators, agitators and emulsifiers constructed and tested worldwide so far have been characterized by low efficiency and high energy demand. Many of them proved effective under laboratory conditions but failed under industrial ones. The only task successfully realized by these appliances and utilized on a wider scale is the heating of liquids. For this reason, their usability was limited to the function of heating installations. Design of the presented cavitation generator allows achieving satisfactory energy efficiency and enables its use under industrial conditions in depolymerization processes of biomass with various characteristics. Investigations conducted on the laboratory and industrial scale confirmed the effectiveness of applying cavitation in the process of biomass destruction. The use of the cavitation generator in laboratory studies for disintegration of sewage sludge allowed increasing biogas production by ca. 30% and shortening the treatment process by ca. 20 - 25%. The shortening of the technological process and increase of wastewater treatment plant effectiveness may delay investments aimed at increasing system output. The use of a mechanical cavitator and application of repeated cavitation process (4-6 times) enables significant acceleration of the biogassing process. In addition, mechanical cavitation accelerates increases in COD and VFA levels.

Keywords: hydrodynamic cavitation, pretreatment, biomass, methane fermentation, Virginia fanpetals

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Authors: Vrushali Guhe, Shailza Singh

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Cutaneous leishmaniasis is neglected tropical disease present worldwide caused by the protozoan parasite Leishmania major, the therapeutic armamentarium for leishmaniasis are showing several limitations as drugs are showing toxic effects with increasing resistance by a parasite. Thus identification of novel therapeutic targets is of paramount importance. Previous studies have shown that autophagy, a cellular process, can either facilitate infection or aid in the elimination of the parasite, depending on the specific parasite species and host background in leishmaniasis. In the present study, our objective was to target the essential autophagy protein ATG8, which plays a crucial role in the survival, infection dynamics, and differentiation of the Leishmania parasite. ATG8 in Leishmania major and its homologue, LC3, in Homo sapiens, act as autophagic markers. Present study manifested the crucial role of ATG8 protein as a potential target for combating Leishmania major infection. Through bioinformatics analysis, we identified non-conserved motifs within the ATG8 protein of Leishmania major, which are not present in LC3 of Homo sapiens. Against these two non-conserved motifs, we generated a peptide library of 60 peptides on the basis of physicochemical properties. These peptides underwent a filtering process based on various parameters, including feasibility of synthesis and purification, compatibility with Selective Reaction Monitoring (SRM)/Multiple reaction monitoring (MRM), hydrophobicity, hydropathy index, average molecular weight (Mw average), monoisotopic molecular weight (Mw monoisotopic), theoretical isoelectric point (pI), and half-life. Further filtering criterion shortlisted three peptides by using molecular docking and molecular dynamics simulations. The direct interaction between ATG8 and the shortlisted peptides was confirmed through Surface Plasmon Resonance (SPR) experiments. Notably, these peptides exhibited the remarkable ability to penetrate the parasite membrane and exert profound effects on Leishmania major. The treatment with these peptides significantly impacted parasite survival, leading to alterations in the cell cycle and morphology. Furthermore, the peptides were found to modulate autophagosome formation, particularly under starved conditions, suggesting their involvement in disrupting the regulation of autophagy within Leishmania major. In vitro, studies demonstrated that the selected peptides effectively reduced the parasite load within infected host cells. Encouragingly, these findings were corroborated by in vivo experiments, which showed a reduction in parasite burden upon peptide administration. Additionally, the peptides were observed to affect the levels of LC3II within host cells. In conclusion, our findings highlight the efficacy of these novel peptides in targeting Leishmania major’s ATG8 and disrupting parasite survival. These results provide valuable insights into the development of innovative therapeutic strategies against leishmaniasis via targeting autophagy protein ATG8 of Leishmania major.

Keywords: ATG8, leishmaniasis, surface plasmon resonance, MD simulation, molecular docking, peptide designing, therapeutics

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Authors: Wisam Ismail, Chole Wright, Elizabeth Baker, Cathy Tait, Mohamed Salhab, Richard Linforth

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Background: Post-mastectomy breast reconstruction is an important treatment option for women with breast cancer with psychosocial, emotional and quality of life benefits. Despite this, Asian patients are one-fifth as likely as Caucasian patients to undergo reconstruction after mastectomy. Aim: This study aimed to assess the difference in breast reconstruction rates between Asian and Caucasian patients treated at Bradford Teaching Hospitals between May 2011 – December 2015.The long-term goal is to equip healthcare professionals to improve breast cancer treatment outcome by increasing breast reconstruction rates in this sub-population. Methods: All patients undergoing mastectomy were identified using a prospectively collected departmental database. Further data was obtained via retrospective electronic case note review. Bradford city population is about 530.000 by the end of 2015, with 67.44% of the city's population was White ethnic groups and 26.83% Asian Ethnic Groups (UK population consensus). The majority of Asian population speaks Urdu, hence an Urdu speaking breast care nurse was appointed to facilitate communications and deliver a better understanding of the reconstruction options and pathways. Statistical analysis was undertaken using the SAS program. Patients were stratified by age, self-reported ethnicity, axillary surgery and reconstruction. Relative odds were calculated using univariate and multivariate logistic regression analyses with adjustment for known confounders. An Urdu speaking breast care nurse was employed throughout this period to facilitate communication and patient decision making. Results: 506 patients underwent Mastectomy over 5 years. 72 (14%) Asian v. 434 (85%) Caucasian. Overall median age is 64 years (SD1.1). Asian median age is 62 (SD0.9), versus Caucasian 65 (SD1.2). Total axillary clearance rate was 30% (42% Asian v.30% Caucasian). Overall reconstruction rate was 126 patients (28.9%).Only 6 of 72 Asian patients (<1%) underwent breast reconstruction versus 121of 434 Caucasian (28%) (p < 0.04), Odds ratio 0.68, (95% confidence interval 0.57-0.79). Conclusions: There is a significant difference in post-mastectomy reconstruction rates between Asian and Caucasian patients. This difference is likely to be multi-factorial. Higher rates of axillary clearance in Asian patients might suggest later disease presentation and/or higher rates of subsequent adjuvant therapy, both of which, can impact on the suitability of breast reconstruction. Strategies aimed at reducing racial disparities in breast reconstruction should include symptom awareness to enable earlier presentation and facilitated communication to ensure informed decision-making.

Keywords: aesthetic, Asian, breast, reconstruction

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Authors: Badryah Alshehri, Kevin Gormley, Gillian Prue, Karen McCutcheon

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In a disaster event, sharing patient information between the pre-hospitals Emergency Medical Services (EMS) and Emergency Department (ED) hospitals is a complex process during which important information may be altered or lost due to poor communication. The aim of this study was to critically discuss the current evidence base in relation to communication between pre-EMS hospital and ED hospital professionals by the use of Information and Communication Systems (ICT). This study followed the systematic approach; six electronic databases were searched: CINAHL, Medline, Embase, PubMed, Web of Science, and IEEE Xplore Digital Library were comprehensively searched in January 2018 and a second search was completed in April 2020 to capture more recent publications. The study selection process was undertaken independently by the study authors. Both qualitative and quantitative studies were chosen that focused on factors which are positively or negatively associated with coordinated communication between pre-hospital EMS and ED teams in a disaster event. These studies were assessed for quality and the data were analysed according to the key screening themes which emerged from the literature search. Twenty-two studies were included. Eleven studies employed quantitative methods, seven studies used qualitative methods, and four studies used mixed methods. Four themes emerged on communication between EMTs (pre-hospital EMS and ED staff) in a disaster event using the ICT. (1) Disaster preparedness plans and coordination. This theme reported that disaster plans are in place in hospitals, and in some cases, there are interagency agreements with pre-hospital and relevant stakeholders. However, the findings showed that the disaster plans highlighted in these studies lacked information regarding coordinated communications within and between the pre-hospital and hospital. (2) Communication systems used in the disaster. This theme highlighted that although various communication systems are used between and within hospitals and pre-hospitals, technical issues have influenced communication between teams during disasters. (3) Integrated information management systems. This theme suggested the need for an integrated health information system which can help pre-hospital and hospital staff to record patient data and ensure the data is shared. (4) Disaster training and drills. While some studies analysed disaster drills and training, the majority of these studies were focused on hospital departments other than EMTs. These studies suggest the need for simulation disaster training and drills, including EMTs. This review demonstrates that considerable gaps remain in the understanding of the communication between the EMS and ED hospitals staff in relation to response in disasters. The review shows that although different types of ICTs are used, various issues remain which affect coordinated communication among the relevant professionals.

Keywords: communication, emergency communication services, emergency medical teams, emergency physicians, emergency nursing, paramedics, information and communication technology, communication systems

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Authors: Eitan Elaad, Yeela Gal-Gonen

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Two studies examined gender differences in lying when the truth-telling bias prevailed and when inspiring lying and distrust. The first study used 156 participants from the community (78 pairs). First, participants completed the Narcissistic Personality Inventory, the Lie- and Truth Ability Assessment Scale (LTAAS), and the Rational-Experiential Inventory. Then, they participated in a deception game where they performed as senders and receivers of true and false communications. Their goal was to retain as many points as possible according to a payoff matrix that specified the reward they would gain for any possible outcome. Results indicated that males in the sender position lied more and were more successful tellers of lies and truths than females. On the other hand, males, as receivers, trusted less than females but were not better at detecting lies and truths. We explained the results by a. Male's high perceived lie-telling ability. We observed that confidence in telling lies guided participants to increase their use of lies. Male's lie-telling confidence corresponded to earlier accounts that showed a consistent association between high self-assessed lying ability, reports of frequent lying, and predictions of actual lying in experimental settings; b. Male's narcissistic features. Earlier accounts described positive relations between narcissism and reported lying or unethical behavior in everyday life situations. Predictions about the association between narcissism and frequent lying received support in the present study. Furthermore, males scored higher than females on the narcissism scale; and c. Male's experiential thinking style. We observed that males scored higher than females on the experiential thinking style scale. We further hypothesized that the experiential thinking style predicts frequent lying in the deception game. Results confirmed the hypothesis. The second study used one hundred volunteers (40 females) who underwent the same procedure. However, the payoff matrix encouraged lying and distrust. Results showed that male participants lied more than females. We found no gender differences in trust. Males and females did not differ in their success of telling and detecting lies and truths. Participants also completed the LTAAS questionnaire. Males assessed their lie-telling ability higher than females, but the ability assessment did not predict lying frequency. A final note. The present design is limited to low stakes. Participants knew that they were participating in a game, and they would not experience any consequences from their deception in the game. Therefore, we advise caution when applying the present results to lying under high stakes.

Keywords: gender, lying, detection of deception, information processing style, self-assessed lying ability

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Authors: André Pasti

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Historically, the communication sector in Argentina is characterized by intense monopolization and geographical concentration in the city of Buenos Aires. In 2000, the four major media conglomerates in operation – Clarín, Telefónica, America and Hadad – controlled 84% of the national media market. By 2009, new policies were implemented as a result of civil society organizations demands. Legally, a new regulatory framework was approved: the law 26,522 of Audiovisual Communications Services. Supposedly, these policies intend to create new conditions for the development of the audiovisual economy in the territory of Argentina. The regionalization of audiovisual production and the democratization of channels and access to media were among the priorities. This paper analyses the main changes and continuities in the organization of the spatial circuit of the audiovisual industry in Argentina provoked by these new policies. These new policies aim at increasing the diversity of audiovisual producers and promoting regional audiovisual industries. For this purpose, a national program for the development of audiovisual centers within the country was created. This program fostered a federalized production network, based on nine audiovisual regions and 40 nodes. Each node has created technical, financial and organizational conditions to gather different actors in audiovisual production – such as SMEs, social movements and local associations. The expansion of access to technical networks was also a concern of other policies, such as ‘Argentina connected’, whose objective was to expand access to broadband Internet. The Open Digital Television network also received considerable investments. Furthermore, measures have been carried out in order to impose limits on the concentration of ownership as well as to eliminate the oligopolies and to ensure more competition in the sector. These actions intended to force a divide of the media conglomerates into smaller groups. Nevertheless, the corporations that compose these conglomerates resist strongly, making full use of their economic and judiciary power. Indeed, the absence of effective impact of such measures can be testified by the fact that the audiovisual industry remains strongly concentrated in Argentina. Overall, these new policies were designed properly to decentralize audiovisual production and expand the regional diversity of the audiovisual industry. However, the effective transformation of the organization of the audiovisual circuit in the territory faced several resistances. This can be explained firstly and foremost by the ideological and economic power of the media conglomerates. In the second place, there is an inherited inertia from the unequal distribution of the objects needed for the audiovisual production and consumption. Lastly, the resistance also relies on financial needs and in the excessive dependence of the state for the promotion of regional audiovisual production.

Keywords: Argentina, audiovisual industry, communication policies, geographic concentration, regionalization, spatial circuit

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Authors: E. Carraro, Sa. Bonetta, Si. Bonetta, E. Ceretti, G. C. V. Viola, C. Pignata, S. Levorato, T. Salvatori, S. Vannini, V. Romanazzi, A. Carducci, G. Donzelli, T. Schilirò, A. De Donno, T. Grassi, S. Bonizzoni, A. Bonetti, G. Gilli, U. Gelatti

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In 2013, air pollution and particulate matter were classified as carcinogenic to human by the IARC. At present, PM is Europe's most problematic pollutant in terms of harm to health, as reported by European Environmental Agency (EEA) in the EEA Technical Report on Air quality in Europe, 2015. A percentage between 17-30 of the EU urban population lives in areas where the EU air quality 24-hour limit value for PM10 is exceeded. Many studies have found a consistent association between exposure to PM and the incidence and mortality for some chronic diseases (i.e. lung cancer, cardiovascular diseases). Among the mechanisms responsible for these adverse effects, genotoxic damage is of particular concern. Children are a high-risk group in terms of the health effects of air pollution and early exposure during childhood can increase the risk of developing chronic diseases in adulthood. The MAPEC_LIFE (Monitoring Air Pollution Effects on Children for supporting public health policy) is a project founded by EU Life+ Programme (LIFE12 ENV/IT/000614) which intends to evaluate the associations between air pollution and early biological effects in children and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children. This work is focused on the micronuclei frequency in child buccal cells in association with airborne PM levels taking into account the influence of other factors associated with the lifestyle of children. The micronucleus test was performed in exfoliated buccal cells of 6–8 years old children from 5 Italian towns with different air pollution levels. Data on air quality during the study period were obtained from the Regional Agency for Environmental Protection. A questionnaire administered to children’s parents was used to obtain details on family socio-economic status, children health condition, exposures to other indoor and outdoor pollutants (i.e. passive smoke) and life-style, with particular reference to eating habits. During the first sampling campaign (winter 2014-15) 1315 children were recruited and sampled for Micronuclei test in buccal cells. In the sampling period the levels of the main pollutants and PM10 were, as expected, higher in the North of Italy (PM10 mean values 62 μg/m3 in Torino and 40 μg/m3 in Brescia) than in the other towns (Pisa, Perugia, Lecce). A higher Micronucleus frequency in buccal cells of children was found in Brescia (0.6/1000 cells) than in the other towns (range 0.3-0.5/1000 cells). The statistical analysis underlines a relation of the micronuclei frequency with PM concentrations, traffic level near child residence, and level of education of parents. The results suggest that, in addition to air pollution exposure, some other factors, related to lifestyle or further exposures, may influence micronucleus frequency and cellular response to air pollutants.

Keywords: air pollution, buccal cells, children, micronucleus cytome assay

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Authors: R. Abdulrahman, A. Eardley, A. Soliman

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The proliferation in the development of mobile learning (m-learning) has played a vital role in the rapidly growing electronic learning market. This relatively new technology can help to encourage the development of in learning and to aid knowledge transfer a number of areas, by familiarizing students with innovative information and communications technologies (ICT). M-learning plays a substantial role in the deployment of learning methods for nursing students by using the Internet and portable devices to access learning resources ‘anytime and anywhere’. However, acceptance of m-learning by students is critical to the successful use of m-learning systems. Thus, there is a need to study the factors that influence student’s intention to use m-learning. This paper addresses this issue. It outlines the outcomes of a study that evaluates the unified theory of acceptance and use of technology (UTAUT) model as applied to the subject of user acceptance in relation to m-learning activity in nurse education. The model integrates the significant components across eight prominent user acceptance models. Therefore, a standard measure is introduced with core determinants of user behavioural intention. The research model extends the UTAUT in the context of m-learning acceptance by modifying and adding individual innovativeness (II) and quality of service (QoS) to the original structure of UTAUT. The paper goes on to add the factors of previous experience (of using mobile devices in similar applications) and the nursing students’ readiness (to use the technology) to influence their behavioural intentions to use m-learning. This study uses a technique called ‘convenience sampling’ which involves student volunteers as participants in order to collect numerical data. A quantitative method of data collection was selected and involves an online survey using a questionnaire form. This form contains 33 questions to measure the six constructs, using a 5-point Likert scale. A total of 42 respondents participated, all from the Nursing Institute at the Armed Forces Hospital in Saudi Arabia. The gathered data were then tested using a research model that employs the structural equation modelling (SEM), including confirmatory factor analysis (CFA). The results of the CFA show that the UTAUT model has the ability to predict student behavioural intention and to adapt m-learning activity to the specific learning activities. It also demonstrates satisfactory, dependable and valid scales of the model constructs. This suggests further analysis to confirm the model as a valuable instrument in order to evaluate the user acceptance of m-learning activity.

Keywords: mobile learning, nursing institute students’ acceptance of m-learning activity in Saudi Arabia, unified theory of acceptance and use of technology model (UTAUT), structural equation modelling (SEM)

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Authors: Dongming Feng, Fangyin Zhang, Liling Cao

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Horizontally curved steel-concrete composite box girders are extensively used in highway bridges. They consist of reinforced concrete deck on top of prefabricated steel box section beam which exhibits a high torsional rigidity to resist torsional effects induced by the curved structural geometry. This type of structural system is often constructed in two stages. The composite section will take the tension mainly by the steel box and, the compression by the concrete deck. The steel girders are delivered in large pre-fabricated U-shaped sections that are designed for ease of construction. They are then erected on site and overlaid by cast-in-place reinforced concrete deck. The functionality of the composite section is not achieved until the closed section is formed by fully cured concrete. Since this kind of composite section is built in two stages, the erection of the open steel box presents some challenges to contractors. When the reinforced concrete slab is cast-in-place, special care should be taken on bracings that can prevent the open U-shaped steel box from global and local buckling. In the case of multiple steel boxes, the design detailing should pay enough attention to the installation requirement of the bracings connecting adjacent steel boxes to prevent the global buckling. The slope in transverse direction and grade in longitudinal direction will result in some local deformation of the steel boxes that affect the connection of the bracings. During the design phase, it is common for engineers to model the curved composite box girder using one-dimensional beam elements. This is adequate to analyze the global behavior, however, it is unable to capture the local deformation which affects the installation of the field bracing connection. The presence of the local deformation may become a critical component to control the construction tolerance, and overlooking this deformation will produce inadequate structural details that eventually cause misalignment in field and erection failure. This paper will briefly describe the construction issues we encountered in real structures, investigate the difference between beam element modeling and shell/solid element modeling, and their impact on the different construction stages. P-delta effect due to the slope and curvature of the composite box girder is analyzed, and the secondary deformation is compared to the first-order response and evaluated for its impact on installation of lateral bracings. The paper will discuss the rational approach to prepare construction documents and recommendations are made on the communications between engineers, erectors, and fabricators to smooth out construction process.

Keywords: buckling, curved composite box girder, stage construction, structural detailing

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Authors: Panagiotis Gkekas, Christos Sougles, Dionysios Kehagias, Dimitrios Tzovaras

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This paper presents the architecture of the “Intelligent cooperative integrated system for road safety and road infrastructure maintenance towards 2020” (ODOS2020) advanced infrastructure, which implements a number of cooperative ITS applications based on Internet of Things and Infrastructure-to-Vehicle (V2I) technologies with the purpose to enhance the active road safety level of vehicles through the provision of a fully automated V2I environment. The primary objective of the ODOS2020 project is to contribute to increased road safety but also to the optimization of time for maintenance of road infrastructure. The integrated technological solution presented in this paper addresses all types of vehicles and requires minimum vehicle equipment. Thus, the ODOS2020 comprises a low-cost solution, which is one of its main benefits. The system architecture includes an integrated notification system to transmit personalized information on road, traffic, and environmental conditions, in order for the drivers to receive real-time and reliable alerts concerning upcoming critical situations. The latter include potential dangers on the road, such as obstacles or road works ahead, extreme environmental conditions, etc., but also informative messages, such as information on upcoming tolls and their charging policies. At the core of the system architecture lies an integrated sensorial network embedded in special road infrastructures (strips) that constantly collect and transmit wirelessly information about passing vehicles’ identification, type, speed, moving direction and other traffic information in combination with environmental conditions and road wear monitoring and predictive maintenance data. Data collected from sensors is transmitted by roadside infrastructure, which supports a variety of communication technologies such as ITS-G5 (IEEE-802.11p) wireless network and Internet connectivity through cellular networks (3G, LTE). All information could be forwarded to both vehicles and Traffic Management Centers (TMC) operators, either directly through the ITS-G5 network, or to smart devices with Internet connectivity, through cloud-based services. Therefore, through its functionality, the system could send personalized notifications/information/warnings and recommendations for upcoming events to both road users and TMC operators. In the course of the ODOS2020 project pilot operation has been conducted to allow drivers of both C-ITS equipped and non-equipped vehicles to experience the provided added value services. For non-equipped vehicles, the provided information is transmitted to a smartphone application. Finally, the ODOS2020 system and infrastructure is appropriate for installation on both urban, rural, and highway environments. The paper presents the various parts of the system architecture and concludes by outlining the various challenges that had to be overcome during its design, development, and deployment in a real operational environment. Acknowledgments: Work presented in this paper was co-financed by the European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation (call RESEARCH–CREATE–INNOVATE) under contract no. Τ1EDK-03081 (project ODOS2020).

Keywords: infrastructure to vehicle, intelligent transportation systems, internet of things, road safety

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Authors: Neyzielle Ronnicque Cadiz

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The Philippine is a hotbed of disasters and is a locus of natural hazards. With an average of 20 typhoons entering the Philippine Area of Responsibility (PAR) each year, seven to eight (7-8) of which makes landfall. The country rather inevitably suffers from climate-related calamities. With this vulnerability to natural hazards, the relevant hazard-related issues that come along with the potential threat and occurrence of a disaster oftentimes garners lesser media attention than when a disaster actually occurred. Post-disaster news and events flood the content of news networks primarily focusing on, but not limited to, the efforts of the national government in resolving post-disaster displacement, and all the more on the community leaders’ incompetence in disaster mitigation-- even though the University of the Philippines’ NOAH Center work hand in hand with different stakeholders for disaster mitigation communication efforts. Disaster risk communication is actually a perennial dilemma. There are so many efforts to reach the grassroots level but emergency and disaster preparedness messages inevitably fall short.. The Philippines is very vulnerable to hazards risk and disasters but social media posts and communication efforts mostly go unnoticed, if not argued upon. This study illustrates the outcomes of a research focusing on the print, broadcast, and social media’s role on disaster communication involving the natural catastrophic events that took place in the Philippines from 2009 to present. Considering the country’s state of development, this study looks on the rapid and reliable communication between the government, and the relief/rescue workers in the affected regions; and how the media portrays these efforts effectively. Learning from the disasters that have occurred in the Philippines over the past decade, effective communication can ensure that any efforts to prepare and respond to disasters can make a significant difference. It can potentially either break or save lives. Recognizing the role of communications is not only in improving the coordination of vital services for post disaster; organizations gave priority in reexamining disaster preparedness mechanisms through the Communication with Communities (CwC) programs. This study, however, looks at the CwC efforts of the Philippine media platforms. CwC, if properly utilized by the media, is an essential tool in ensuring accountability and transparency which require effective exchange of information between disasters and survivors and responders. However, in this study, it shows that the perennial dilemma of the Philippine media is that the Disaster Risk Reduction and Management (DRRM) efforts of the country lie in the clouded judgment of political aims. This kind of habit is a multiplier of the country’s risk and insecurity. Sometimes the efforts in urging the public to take action seem useless because the challenge lies on how to achieve social, economic, and political unity using the tri-media platform.

Keywords: Philippines at risk, pre/post disaster communication, tri-media platform, UP NOAH

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Authors: Inna Kornienko, Svetlana Guryeva, Anatoly Shekhter, Elena Petersen

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Transplantation is an effective treatment option for patients suffering from different end-stage diseases. However, it is plagued by a constant shortage of donor organs and the subsequent need of a lifelong immunosuppressive therapy for the patient. Currently some researchers look towards using of pig organs to replace human organs for transplantation since the matrix derived from porcine organs is a convenient substitute for the human matrix. As an initial step to create a new ex vivo tissue engineered model, optimized protocols have been created to obtain organ-specific acellular matrices and evaluated their potential as tissue engineered scaffolds for culture of normal cells and tumor cell lines. These protocols include decellularization by perfusion in a bioreactor system and immersion-agitation on an orbital shaker with use of various detergents (SDS, Triton X-100) and freezing. Complete decellularization – in terms of residual DNA amount – is an important predictor of probability of immune rejection of materials of natural origin. However, the signs of cellular material may still remain within the matrix even after harsh decellularization protocols. In this regard, the matrices obtained from tissues of low-immunogenic pigs with α3Galactosyl-tranferase gene knock out (GalT-KO) may be a promising alternative to native animal sources. The research included a study of induced effect of frozen and fresh fragments of GalT-KO skin on healing of full-thickness plane wounds in 80 rats. Commercially available wound dressings (Ksenoderm, Hyamatrix and Alloderm) as well as allogenic skin were used as a positive control and untreated wounds were analyzed as a negative control. The results were evaluated on the 4th day after grafting, which corresponds to the time of start of normal wound epithelization. It has been shown that a non-specific immune response in models treated with GalT-Ko pig skin was milder than in all the control groups. Research has been performed to measure technical skin characteristics: stiffness and elasticity properties, corneometry, tevametry, and cutometry. These metrics enabled the evaluation of hydratation level, corneous layer husking level, as well as skin elasticity and micro- and macro-landscape. These preliminary data may contribute to development of personalized transplantable organs from GalT-Ko pigs with significantly limited potential of immune rejection. By applying growth factors to a decellularized skin sample it is possible to achieve various regenerative effects based on the particular situation. In this particular research BMP2 and Heparin-binding EGF-like growth factor have been used. Ideally, a bioengineered organ must be biocompatible, non-immunogenic and support cell growth. Porcine organs are attractive for xenotransplantation if severe immunologic concerns can be bypassed. The results indicate that genetically modified pig tissues with knock-outed α3Galactosyl-tranferase gene may be used for production of low-immunogenic matrix suitable for transplantation.

Keywords: decellularization, low-immunogenic, matrix, scaffolds, transplants

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Authors: Aberdam Edith, Sangari Linda, Petit Isabelle, Aberdam Daniel

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Background and Rationale: Transparent avascularised cornea is essential for normal vision and depends on limbal stem cells (LSC) that reside between the cornea and the conjunctiva. Ocular burns or injuries may destroy the limbus, causing limbal stem cell deficiency (LSCD). The cornea becomes vascularised by invaded conjunctival cells, the stroma is scarring, resulting in corneal opacity and loss of vision. Grafted autologous limbus or cultivated autologous LCS can restore the vision, unless the two eyes are affected. Alternative cellular sources have been tested in the last decades, including oral mucosa or hair follicle epithelial cells. However, only partial success has been achieved by the use of these cells since they were not able to uniformly commit into corneal epithelial cells. Human pluripotent stem cells (iPSC) display both unlimited growth capacity and ability to differentiate into any cell type. Our goal was to design a standardized and reproducible protocol to produce transplantable autologous LSC from patients through cell reprogramming technology. Methodology: First, keratinocyte primary culture was established from a small number of plucked hair follicles of healthy donors. The resulting epithelial cells were reprogrammed into induced pluripotent stem cells (iPSCs) and further differentiate into corneal epithelial cells (CEC), according to a robust protocol that recapitulates the main step of corneal embryonic development. qRT-PCR analysis and immunofluorescent staining during the course of differentiation confirm the expression of stage specific markers of corneal embryonic lineage. First appear ectodermal progenitor-specific cytokeratins K8/K18, followed at day 7 by limbal-specific PAX6, TP63 and cytokeratins K5/K14. At day 15, K3/K12+-corneal cells are present. To amplify the iPSC-derived LSC (named COiPSC), intact small epithelial colonies were detached and cultivated in limbal cell-specific medium. In that culture conditions, the COiPSC can be frozen and thaw at any passage, while retaining their corneal characteristics for at least eight passages. To evaluate the potential of COiPSC as an alternative ocular toxicity model, COiPSC were treated at passage P0 to P4 with increasing amounts of SDS and Benzalkonium. Cell proliferation and apoptosis of treated cells was compared to LSC and the SV40-immortalized human corneal epithelial cell line (HCE) routinely used by cosmetological industrials. Of note, HCE are more resistant to toxicity than LSC. At P0, COiPSC were systematically more resistant to chemical toxicity than LSC and even to HCE. Remarkably, this behavior changed with passage since COiPSC at P2 became identical to LSC and thus closer to physiology than HCE. Comparative transcriptome analysis confirmed that COiPSC from P2 are similar to a mixture of LSC and CEC. Finally, by organotypic reconstitution assay, we demonstrated the ability of COiPSC to produce a 3D corneal epithelium on a stromal equivalent made of keratocytes. Conclusion: COiPSC could become valuable for two main applications: (1) an alternative robust tool to perform, in a reproducible and physiological manner, toxicity assays for cosmetic products and pharmacological tests of drugs. (2). COiPSC could become an alternative autologous source for cornea transplantation for LSCD.

Keywords: Limbal stem cell deficiency, iPSC, cornea, limbal stem cells

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Authors: Mohammadjavad Sotoudeheian

Abstract:

Heart failure (HF) prevalence, as a global cardiovascular problem, is increasing gradually. A variety of molecular mechanisms contribute to HF. Proteins involved in cardiac contractility regulation, such as ion channels and calcium handling proteins, are altered. Additionally, epigenetic modifications and gene expression can lead to altered cardiac function. Moreover, inflammation and oxidative stress contribute to HF. The progression of HF can be attributed to mitochondrial dysfunction that impairs energy production and increases apoptosis. Molecular mechanisms such as these contribute to the development of cardiomyocyte defects and HF and can be therapeutically targeted. The heart's contractile function is controlled by cardiomyocytes. Calpain, and its related molecules, including Bax, VEGF, and AMPK, are among the proteins involved in regulating cardiomyocyte function. Apoptosis is facilitated by Bax. Cardiomyocyte apoptosis is regulated by this protein. Furthermore, cardiomyocyte survival, contractility, wound healing, and proliferation are all regulated by VEGF, which is produced by cardiomyocytes during inflammation and cytokine stress. Cardiomyocyte proliferation and survival are also influenced by AMPK, an enzyme that plays an active role in energy metabolism. They all play key roles in apoptosis, angiogenesis, hypertrophy, and metabolism during myocardial inflammation. The role of calpains has been linked to several molecular pathways. The calpain pathway plays an important role in signal transduction and apoptosis, as well as autophagy, endocytosis, and exocytosis. Cell death and survival are regulated by these calcium-dependent cysteine proteases that cleave proteins. As a result, protein fragments can be used for various cellular functions. By cleaving adhesion and motility proteins, calcium proteins also contribute to cell migration. HF may be brought about by calpain-mediated pathways. Many physiological processes are mediated by the calpain molecular pathways. Signal transduction, cell death, and cell migration are all regulated by these molecular pathways. Calpain is activated by calcium binding to calmodulin. In the presence of calcium, calmodulin activates calpain. Calpains are stimulated by calcium, which increases matrix metalloproteinases (MMPs). In order to develop novel treatments for these diseases, we must understand how this pathway works. A variety of myocardial remodeling processes involve calpains, including remodeling of the extracellular matrix and hypertrophy of cardiomyocytes. Calpains also play a role in maintaining cardiac homeostasis through apoptosis and autophagy. The development of HF may be in part due to calpain-mediated pathways promoting cardiomyocyte death. Numerous studies have suggested the importance of the Ca2+ -dependent protease calpain in cardiac physiology and pathology. Therefore, it is important to consider this pathway to develop and test therapeutic options in humans that targets calpain in HF. Apoptosis, autophagy, endocytosis, exocytosis, signal transduction, and disease progression all involve calpain molecular pathways. Therefore, it is conceivable that calpain inhibitors might have therapeutic potential as they have been investigated in preclinical models of several conditions in which the enzyme has been implicated that might be treated with them. Ca 2+ - dependent proteases and calpains contribute to adverse ventricular remodeling and HF in multiple experimental models. In this manuscript, we will discuss the calpain molecular pathway's important roles in HF development.

Keywords: calpain, heart failure, autophagy, apoptosis, cardiomyocyte

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Authors: Ekaterina Zvorykina

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Nowadays, Arctic and Antarctic regions are distinguished to be one of the most important strategic resources for global development. Nonetheless, living conditions in Arctic regions still demand certain improvements. As soon as the region is rarely populated, one of the main points of interest is health accommodation of the people, who migrate to Arctic region for permanent and shift work. At Arctic and Antarctic latitudes, personnel face polar day and polar night conditions during the time of the year. It means that they are deprived of natural sunlight in winter season and have continuous daylight in summer. Firstly, the change in light intensity during 24-hours period due to migration affects circadian rhythms. Moreover, the controlled artificial light in winter is also an issue. The results of the recent studies on night shift medical professionals, who were exposed to permanent artificial light, have already demonstrated higher risks in cancer, depression, Alzheimer disease. Moreover, people exposed to frequent time zones change are also subjected to higher risks of heart attack and cancer. Thus, our main goals are to understand how high latitude work and living conditions can affect human health and how it can be prevented. In our study, we analyze molecular and cellular factors, which play important role in circadian rhythm change and distinguish main risk groups in people, migrating to high latitudes. The main well-studied index of circadian timing is melatonin or its metabolite 6-sulfatoxymelatonin. In low light intensity melatonin synthesis is disturbed and as a result human organism requires more time for sleep, which is still disregarded when it comes to working time organization. Lack of melatonin also causes shortage in serotonin production, which leads to higher depression risk. Melatonin is also known to inhibit oncogenes and increase apoptosis level in cells, the main factors for tumor growth, as well as circadian clock genes (for example Per2). Thus, people who work in high latitudes can be distinguished as a risk group for cancer diseases and demand more attention. Clock/Clock genes, known to be one of the main circadian clock regulators, decrease sensitivity of hypothalamus to estrogen and decrease glucose sensibility, which leads to premature aging and oestrous cycle disruption. Permanent light exposure also leads to accumulation superoxide dismutase and oxidative stress, which is one of the main factors for early dementia and Alzheimer disease. We propose a new screening system adjusted for people, migrating from middle to high latitudes and accommodation therapy. Screening is focused on melatonin and estrogen levels, sleep deprivation and neural disorders, depression level, cancer risks and heart and vascular disorders. Accommodation therapy includes different types artificial light exposure, additional melatonin and neuroprotectors. Preventive procedures can lead to increase of migration intensity to high latitudes and, as a result, the prosperity of Arctic region.

Keywords: circadian rhythm, high latitudes, melatonin, neuroprotectors

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Authors: Ricardo Alberto Chiong Zevallos, Eduardo Moraes Rego Reis

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Pancreatic adenocarcinoma (PDAC) patients have a less than 10% 5-year survival rate. PDAC has no defined diagnostic and prognostic biomarkers. Gemcitabine is the first-line drug in PDAC and several other cancers. Long non-coding RNAs (lncRNAs) contribute to the tumorigenesis and are potential biomarkers for PDAC. Although lncRNAs aren’t translated into proteins, they have important functions. LncRNAs can decoy or recruit proteins from the epigenetic machinery, act as microRNA sponges, participate in protein translocation through different cellular compartments, and even promote chemoresistance. The chromatin remodeling enzyme EZH2 is a histone methyltransferase that catalyzes the methylation of histone 3 at lysine 27, silencing local expression. EZH2 is ambivalent, it can also activate gene expression independently of its histone methyltransferase activity. EZH2 is overexpressed in several cancers and interacts with lncRNAs, being recruited to a specific locus. EZH2 can be recruited to activate an oncogene or silence a tumor suppressor. The lncRNAs misregulation in cancer can result in the differential recruitment of EZH2 and in a distinct epigenetic landscape, promoting chemoresistance. The relevance of the EZH2-lncRNAs interaction to chemoresistant PDAC was assessed by Real Time quantitative PCR (RT-qPCR) and RNA Immunoprecipitation (RIP) experiments with naïve and gemcitabine-resistant PDAC cells. The expression of several lncRNAs and EZH2 gene targets was evaluated contrasting naïve and resistant cells. Selection of candidate genes was made by bioinformatic analysis and literature curation. Indeed, the resistant cell line showed higher expression of chemoresistant-associated lncRNAs and protein coding genes. RIP detected lncRNAs interacting with EZH2 with varying intensity levels in the cell lines. During RIP, the nuclear fraction of the cells was incubated with an antibody for EZH2 and with magnetic beads. The RNA precipitated with the beads-antibody-EZH2 complex was isolated and reverse transcribed. The presence of candidate lncRNAs was detected by RT-qPCR, and the enrichment was calculated relative to INPUT (total lysate control sample collected before RIP). The enrichment levels varied across the several lncRNAs and cell lines. The EZH2-lncRNA interaction might be responsible for the regulation of chemoresistance-associated genes in multiple cancers. The relevance of the lncRNA-EZH2 interaction to PDAC was assessed by siRNA knockdown of a lncRNA, followed by the analysis of the EZH2 target expression by RT-qPCR. The chromatin immunoprecipitation (ChIP) of EZH2 and H3K27me3 followed by RT-qPCR with primers for EZH2 targets also assess the specificity of the EZH2 recruitment by the lncRNA. This is the first report of the interaction of EZH2 and lncRNAs HOTTIP and PVT1 in chemoresistant PDAC. HOTTIP and PVT1 were described as promoting chemoresistance in several cancers, but the role of EZH2 is not clarified. For the first time, the lncRNA LINC01133 was detected in a chemoresistant cancer. The interaction of EZH2 with LINC02577, LINC00920, LINC00941, and LINC01559 have never been reported in any context. The novel lncRNAs-EZH2 interactions regulate chemoresistant-associated genes in PDAC and might be relevant to other cancers. Therapies targeting EZH2 alone weren’t successful, and a combinatorial approach also targeting the lncRNAs interacting with it might be key to overcome chemoresistance in several cancers.

Keywords: epigenetics, chemoresistance, long non-coding RNAs, pancreatic cancer, histone modification

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