Search results for: neuronal differentiation

Authors: Jung Park, Anca Mazare, Klaus Von Der Mark, Patrik Schmuki

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In clinical application of TiO2 implants on tooth and hip replacement, migration, adhesion and differentiation of neighboring mesenchymal stem cells onto implant surfaces are critical steps for successful bone regeneration. In a recent decade, accumulated attention has been paid on nanoscale electrochemical surface modifications on TiO2 layer for improving bone-TiO2 surface integration. We generated, on titanium surfaces, self-assembled layers of vertically oriented TiO2 nanotubes with defined diameters between 15 and 100 nm and here we show that mesenchymal stem cells finely sense TiO2 nanotubular geometry and quickly decide their cell fate either to differentiation into osteoblasts or to programmed cell death (apoptosis) on TiO2 nanotube layers. These cell fate decisions are critically dependent on nanotube size differences (15-100nm in diameters) of TiO2 nanotubes sensing by integrin clustering. We further demonstrate that nanoscale topography-sensing is feasible not only in mesenchymal stem cells but rather seems as generalized nanoscale microenvironment-cell interaction mechanism in several cell types composing bone tissue network including osteoblasts, osteoclast, endothelial cells and hematopoietic stem cells. Additionally we discuss the synergistic effect of simultaneous stimulation by nanotube-bound growth factor and nanoscale topographic cues on enhanced bone regeneration.

Keywords: TiO2 nanotube, stem cell fate decision, nano-scale microenvironment, bone regeneration

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Authors: Ghada E. Esheba, Ghadeer F. Alharthi, Duaa A. Alhejaili, Rawan E. Hudairy, Wafaa A. Altaezi, Raghad M. Alhejaili

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Background: Inflammatory bowel disease (IBD) consists of two specific gastrointestinal disorders: ulcerative colitis (UC) and Crohn's disease (CD). Despite their distinct natures, these two diseases share many similar etiologic, clinical and pathological features, as a result, their accurate differential diagnosis may sometimes be difficult. Correct diagnosis is important because surgical treatment and long-term prognosis differ from UC and CD. Aim: This study aims to study the characteristic clinicopathological features which help in the differential diagnosis between UC and CD, and assess the disease activity in ulcerative colitis. Materials and methods: This study was carried out on 50 selected cases. The cases included 27 cases of UC and 23 cases of CD. All the cases were examined using H& E and immunohistochemically for bcl-2 expression. Results: Characteristic features of UC include: decrease in mucous content, irregular or villous surface, crypt distortion, and cryptitis, whereas the main cardinal histopathological features seen in CD were: epitheloid granuloma, transmural chronic inflammation, absence of mucin depletion, irregular surface, or crypt distortion. 3 cases of UC were found to be associated with dysplasia. UC mucosa contains fewer Bcl-2+ cells compared with CD mucosa. Conclusion: This study using multiple parameters such clinicopathological features and Bcl-2 expression as studied by immunohistochemical stain, helped to gain an accurate differentiation between UC and CD. Furthermore, this work spotted the light on the activity and different grades of UC which could be important for the prediction of relapse.

Keywords: Crohn's disease, dysplasia, inflammatory bowel disease, ulcerative colitis

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Authors: Zeytu G. Asfaw, Serkalem K. Abrha, Demisew G. Degefu

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Background: HIV/AIDS remains a major public health problem in Ethiopia and heavily affecting people of productive and reproductive age. We aimed to compare the performance of Parametric Survival Analysis and Bayesian Survival Analysis using simulations and in a real dataset application focused on determining predictors of HIV patient survival. Methods: A Parametric Survival Models - Exponential, Weibull, Log-normal, Log-logistic, Gompertz and Generalized gamma distributions were considered. Simulation study was carried out with two different algorithms that were informative and noninformative priors. A retrospective cohort study was implemented for HIV infected patients under Highly Active Antiretroviral Therapy in Alamata General Hospital, North Ethiopia. Results: A total of 320 HIV patients were included in the study where 52.19% females and 47.81% males. According to Kaplan-Meier survival estimates for the two sex groups, females has shown better survival time in comparison with their male counterparts. The median survival time of HIV patients was 79 months. During the follow-up period 89 (27.81%) deaths and 231 (72.19%) censored individuals registered. The average baseline cluster of differentiation 4 (CD4) cells count for HIV/AIDS patients were 126.01 but after a three-year antiretroviral therapy follow-up the average cluster of differentiation 4 (CD4) cells counts were 305.74, which was quite encouraging. Age, functional status, tuberculosis screen, past opportunistic infection, baseline cluster of differentiation 4 (CD4) cells, World Health Organization clinical stage, sex, marital status, employment status, occupation type, baseline weight were found statistically significant factors for longer survival of HIV patients. The standard error of all covariate in Bayesian log-normal survival model is less than the classical one. Hence, Bayesian survival analysis showed better performance than classical parametric survival analysis, when subjective data analysis was performed by considering expert opinions and historical knowledge about the parameters. Conclusions: Thus, HIV/AIDS patient mortality rate could be reduced through timely antiretroviral therapy with special care on the potential factors. Moreover, Bayesian log-normal survival model was preferable than the classical log-normal survival model for determining predictors of HIV patients survival.

Keywords: antiretroviral therapy (ART), Bayesian analysis, HIV, log-normal, parametric survival models

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Authors: Changdong Oh

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With the arrival of post-industrial society, social scientists have been giving attention to issues of which factors shape cleavage of political parties. Especially, there is a heated controversy over whether and how social and cultural values influence the identities of parties and voting behavior. Drawing from Moral Foundations Theory (MFT), which approached similar issues by considering the effect of five moral foundations on political decision-making of people, this study investigates the role of moral rhetoric in the evolution of Korean political parties. Researcher collected official announcements released by the major two parties (Democratic Party of Korea, Saenuri Party) from 2007 to 2016, and analyzed the data by using Word2Vec algorithm and Moral Foundations Dictionary. Five moral decision modules of MFT, composed of care, fairness (individualistic morality), loyalty, authority and sanctity (group-based, Durkheimian morality), can be represented in vector spaces consisted of party announcements data. By comparing the party vector and the five morality vectors, researcher can see how the political parties have actively used each of the five moral foundations to express themselves and the opposition. Results report that the conservative party tends to actively draw on collective morality such as loyalty, authority, purity to differentiate itself. Notably, such moral differentiation strategy is prevalent when they criticize an opposition party. In contrast, the liberal party tends to concern with individualistic morality such as fairness. This result indicates that moral cleavage does exist between parties in South Korea. Furthermore, individualistic moral gaps of the two political parties are eased over time, which seems to be due to the discussion of economic democratization of conservative party that emerged after 2012, but the community-related moral gaps widened. These results imply that past political cleavages related to economic interests are diminishing and replaced by cultural and social values associated with communitarian morality. However, since the conservative party’s differentiation strategy is largely related to negative campaigns, it is doubtful whether such moral differentiation among political parties can contribute to the long-term party identification of the voters, thus further research is needed to determine it is sustainable. Despite the limitations, this study makes it possible to track and identify the moral changes of party system through automated text analysis. More generally, this study could contribute to the analysis of various texts associated with the moral foundation and finding a distributed representation of moral, ethical values.

Keywords: moral foundations theory, moral politics, party system, Word2Vec

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Authors: Lin Zhang

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The fear of leprosy has been longstanding throughout the human history. In an era where isolation is practiced as a means of epidemic prevention, the leprosy rehabilitation group has itself become an isolated community with an entrenched metaphor. In the process of new mediatization of the leprosy isolation community, what are the relations among media literacy, the leprosy internalized stigma and social support? To address the question, the “portrait” of leprosy rehabilitation group is re-delineated through two field studies in the “post-leprosy age” in 2012 and 2020, respectively. Taking an isolation community on Si’an Leprosy Island in Dongguan City, Guangdong Province, China as the study object, it is found that new media promotes the dissolution of segregation effect of the leprosy isolation community and the cultivation of opinion leaders by breaking spatial, psychological and social segregation and by building a community of village affairs and public space in the following way: the cured patients with high new media literacy, especially those who use WeChat and other applications and largely rely on new media for information, have a low level of leprosy internalized stigma and a high level of social support, and they are often the opinion leaders inside their community; on the contrary, the cured patients with low new media literacy, a high level of leprosy internalized stigma and a low level of social support are often the followers inside their community. Such effects of dissolution and construction are reflected not only in the vertical differentiation of the same individual at different times, but also in the horizontal differentiation between different individuals at the same time.

Keywords: segregation, the leprosy rehabilitation group, new mediatization, digital empowerment, opinion leaders

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Authors: A. Reum Son, Jin Seon Kwon, Seung Hun Park, Hai Bang Lee, Moon Suk Kim

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These days, stem cell therapy is focused on for promising source of treatment in clinical human disease. As a supporter of stem cells, in situ-forming hydrogels with growth factors and cells appear to be a promising approach in tissue engineering. To examine osteogenic differentiation of hTMSCs which is one of mesenchymal stem cells in vivo in an injectable hydrogel, we use a methoxy polyethylene glycol-polycaprolactone blockcopolymer (MPEG-PCL) solution with osteogenic factors. We synthesized MPEG-PCL hydrogel and measured viscosity to check sol-gel transition. In order to demonstrate osteogenic ability of hTMSCs, we conducted in vitro osteogenesis experiment. Then, to confirm the cell cytotoxicity, we performed WST-1 with hTMSCs and MPEG-PCL. As the result of in vitro experiment, we implanted cell and hydrogel mixture into animal model and checked degree of osteogenesis with histological analysis and amount of expression genes. Through these experimental data, MPEG-PCL hydrogel has sol-gel transition in temperature change and is biocompatible with stem cells. In histological analysis and gene expression, hTMSCs are very good source of osteogenesis with hydrogel and will use it to tissue engineering as important treatment method. hTMSCs could be a good adult stem cell source for usability of isolation and high proliferation. When hTMSCs are used as cell therapy method with in situ-formed hydrogel, they may provide various benefits like a noninvasive alternative for bone tissue engineering applications.

Keywords: injectable hydrogel, stem cell, osteogenic differentiation, tissue engineering

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Authors: Beate Englich, Linda Schlittenbauer, Christiane Pfeifer, Isabel Kratochvil, Michael Borte, Gabriele I. Stangl, Martin von Bergen, Thorsten Reemtsma, Irina Lehmann, Kristin M. Junge

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The worldwide production of endocrine disrupting compounds (EDC) has risen dramatically over the last decades, as so has the prevalence for obesity. Many EDCs are believed to contribute to this obesity epidemic, by enhancing adipogenesis or disrupting relevant metabolism. This effect is most tremendous in the early prenatal period when priming effects find a highly vulnerable time window. Therefore, we investigate the impact of parabens on childhood overweight development and adipogenesis in general. Parabens are ester of 4-hydroxy-benzoic acid and part of many cosmetic products or food packing. Therefore, ubiquitous exposure can be found in the westernized world, with exposure already starting during the sensitive prenatal period. We assessed maternal cosmetic product consumption, prenatal paraben exposure and infant BMI z-scores in the prospective German LINA cohort. In detail, maternal urinary concentrations (34 weeks of gestation) of methyl paraben (MeP), ethyl paraben (EtP), n-propyl paraben (PrP) and n-butyl paraben (BuP) were quantified using UPLC-MS/MS. Body weight and height of their children was assessed during annual clinical visits. Further, we investigated the direct influence of those parabens on adipogenesis in-vitro using a human mesenchymal stem cell (MSC) differentiation assay to mimic a prenatal exposure scenario. MSC were exposed to 0.1 – 50 µM paraben during the entire differentiation period. Differentiation outcome was monitored by impedance spectrometry, real-time PCR and triglyceride staining. We found that maternal cosmetic product consumption was highly correlated with urinary paraben concentrations at pregnancy. Further, prenatal paraben exposure was linked to higher BMI Z-scores in children. Our in-vitro analysis revealed that especially the long chained paraben BuP stimulates adipogenesis by increasing the expression of adipocyte specific genes (PPARγ, ADIPOQ, LPL, etc.) and triglyceride storage. Moreover, we found that adiponectin secretion is increased whereas leptin secretion is reduced under BuP exposure in-vitro. Further mechanistic analysis for receptor binding and activation of PPARγ and other key players in adipogenesis are currently in process. We conclude that maternal cosmetic product consumption is linked to prenatal paraben exposure of children and contributes to the development of infant overweight development by triggering key pathways of adipogenesis.

Keywords: adipogenesis, endocrine disruptors, paraben, prenatal exposure

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Authors: Rafia S. Al-lamki, Jun Wang, Simon Pacey, Jordan Pober, John R. Bradley

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Tumor necrosis factor alpha (TNF), signaling through TNFR2, may act an autocrine growth factor for renal tubular epithelial cells. Clear cell renal carcinomas (ccRCC) contain cancer stem cells (CSCs) that give rise to progeny which form the bulk of the tumor. CSCs are rarely in cell cycle and, as non-proliferating cells, resist most chemotherapeutic agents. Thus, recurrence after chemotherapy may result from the survival of CSCs. Therapeutic targeting of both CSCs and the more differentiated bulk tumor populations may provide a more effective strategy for treatment of RCC. In this study, we hypothesized that TNFR2 signaling will induce CSCs in ccRCC to enter cell cycle so that treatment with ligands that engage TNFR2 will render CSCs susceptible to chemotherapy. To test this hypothesis, we have utilized wild-type TNF (wtTNF) or specific muteins selective for TNFR1 (R1TNF) or TNFR2 (R2TNF) to treat either short-term organ cultures of ccRCC and adjacent normal kidney (NK) tissue or cultures of CD133+ cells isolated from ccRCC and adjacent NK, hereafter referred to as stem cell-like cells (SCLCs). The effect of cyclophosphamide (CP), currently an effective anticancer agent, was tested on CD133+SCLCs from ccRCC and NK before and after R2TNF treatment. Responses to TNF were assessed by flow cytometry (FACS), immunofluorescence, and quantitative real-time PCR, TUNEL, and cell viability assays. Cytotoxic effect of CP was analyzed by Annexin V and propidium iodide staining with FACS. In addition, we assessed the effect of TNF on isolated SCLCs differentiation using a three-dimensional (3D) culture system. Clinical samples of ccRCC contain a greater number SCLCs compared to NK and the number of SCSC increases with higher tumor grade. Isolated SCLCs show expression of stemness markers (oct4, Nanog, Sox2, Lin28) but not differentiation markers (cytokeratin, CD31, CD45, and EpCAM). In ccRCC organ cultures, wtTNF and R2TNF increase CD133 and TNFR2 expression and promote cell cycle entry whereas wtTNF and R1TNF increase TNFR1 expression and promote cell death of SCLCs. Similar findings are observed in SCLCs isolated from NK but the effect was greater in SCLCs isolated from ccRCC. Application of CP distinctly triggered apoptotic and necrotic cell death in SLCSs pre-treatment with R2TNF as compared to CP treatment alone, with SCLCs from ccRCC more sensitive to CP compared to SLCS from NK. Furthermore, TNF promotes differentiation of SCLCs to an epithelial phenotype in 3D cultures, confirmed by cytokeratin expression and loss of stemness markers Nanog and Sox2. The differentiated cells show positive expression of TNF and TNFR2. These findings provide evidence that selective engagement of TNFR2 drive CSCs to cell proliferation/differentiation, and targeting of cycling cells with TNFR2 agonist in combination with anti-cancer agents may be a potential therapy for RCC.

Keywords: cancer stem cells, ccRCC, cell cycle, cell death, TNF, TNFR1, TNFR2, CD133

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Authors: Samira Baghbanbari, A. B. P. Lever, Payam S. Shabestari, Donald Weaver

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Existing signal transmission models, although undoubtedly useful, have proven insufficient to explain the full complexity of information transfer within the central nervous system. The development of transformative models will necessitate a more comprehensive understanding of neuronal lipid membrane electrophysiology. Pursuant to this goal, the role of highly organized interfacial phospholipid-water layers emerges as a promising case study. A series of phospholipids in neural-glial gap junction interfaces as well as cholesterol molecules have been computationally modelled using high-performance density functional theory (DFT) calculations. Subsequent 'charge decomposition analysis' calculations have revealed a net transfer of charge from phospholipid orbitals through the organized interfacial water layer before ultimately finding its way to cholesterol acceptor molecules. The specific pathway of charge transfer from phospholipid via water layers towards cholesterol has been mapped in detail. Cholesterol is an essential membrane component that is overrepresented in neuronal membranes as compared to other mammalian cells; given this relative abundance, its apparent role as an electronic acceptor may prove to be a relevant factor in further signal transmission studies of the central nervous system. The timescales over which this electronic charge transfer occurs have also been evaluated by utilizing a system design that systematically increases the number of water molecules separating lipids and cholesterol. Memory loss through hydrogen-bonded networks in water can occur at femtosecond timescales, whereas existing action potential-based models are limited to micro or nanosecond scales. As such, the development of future models that attempt to explain faster timescale signal transmission in the central nervous system may benefit from our work, which provides additional information regarding fast timescale energy transfer mechanisms occurring through interfacial water. The study possesses a dataset that includes six distinct phospholipids and a collection of cholesterol. Ten optimized geometric characteristics (features) were employed to conduct binary classification through an artificial neural network (ANN), differentiating cholesterol from the various phospholipids. This stems from our understanding that all lipids within the first group function as electronic charge donors, while cholesterol serves as an electronic charge acceptor.

Keywords: charge transfer, signal transmission, phospholipids, water layers, ANN

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Authors: Erik Kuravsky

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One of the problems with a post-structuralist account of agency is that it appears to reject the freedom of an acting subject, thus seeming to deny the very phenomenon of agency. However, this is only a problem if we think that human beings can be agents exclusively in terms of being subjects, that is, if we think agency subjectively. Indeed, we tend to understand traditional theories of human freedom (e.g., Plato’s or Kant’s) in terms of a peculiar ability of the subject. The paper suggests to de-subjectivize agency with the help of Heidegger’s later thought. To do it, ir argues that classical theories of agency may indeed be interpreted as subject-oriented (sometimes even by their authors), but do not have to be read as such. Namely, the claim is that what makes agency what it is, what is essential in agency, is not its belonginess to a subject, but its ontological configuration. We may say that agency “happens,” and that there is a very specific ontological characteristics to this happening. The argument of the paper is that we can find these characteristic in the classical accounts of agency and that these characteristics are sufficient to distinguish human freedom from other natural phenomena. In particular, it offers to think agency not as one of human characteristics, but as an ontological event in which human beings take part. Namely, agency is a (non-human) characteristic of the different modes in which the experienceable existence of beings is determined by Being. To be an agent then is to participate in such ontological determination. What enables this participation is the ways human beings non-thematically understand the ontological difference. For example, for Plato, one acts freely only if one is led by an idea of the good, while for Kant the imperative for free action is categorial. The agency of an agent is thus dependent on the differentiation between ideas/categories and beings met in experience – one is “free” from contingent sensibility in terms of what is different from it ontologically. In this light, modern dependence on subjectivity is evident in the fact that the ontological difference is thought as belonging to one’s thinking, consciousness etc. That is, it is taken subjectively. A non-subjective account of agency, on the other hand, requires thinking this difference as belonging to Being itself, and thinking human beings as a medium within which occurs the non-human force of ontological differentiation.

Keywords: Heidegger, freedom, agency, poststructuralism

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Authors: Karla Kristine Bay, Jovie Ann Gabin, Allana Joyce Sasotona

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This research employed an Interpretative Phenomenological Analysis to explore the experiences of Children in Conflict with the Law (CICL) which gave light to their concept of ‘friends’. Derived from this context are the following objectives of the study: 1) determining the differentiation of the forms of friends of the CICL; 2) presenting the process of attachment towards detachment in the formation of friendship; and 3) discussing the experiences, and reflections of the CICL on the ‘self’ out of their encounter with friendship. Using the data gathered from the individual drawings of the CICL of their representations of the self, family, friends, community, and Bahay Kalinga as subjects in the meaning-making process utilizing Filipino Psychology methods of pagtatanong-tanong (interview), and pakikipagkwentuhan (conversation), data analysis produced a synthesis of seventeen individual cases. Overall results generated three superordinate themes on the differentiation of the forms of friends which include friends with good influences, friends with bad influences, and friends within the family. While two superordinate themes were produced on the process of attachment towards detachment, namely social, emotional, and psychological experiences on the process of attachment, and emotional and psychological experiences on the process of detachment. Lastly, two superordinate themes were created on the experiences, and reflections of the CICL on the ‘self’ out of their encounter with friendship. This consists of the recognition of the ‘self’ as a responsible agent in developing healthy relationships between the self and others, and reconstruction of the self from the collective experiences of healing, forgiveness, and acceptance. These findings, together with supporting theories discussed the impact of friendship on the emergence of criminal behavior and other dispositions; springing from the child’s dissociation from the family that led to finding belongingness from an external group called friends.

Keywords: children in conflict with the law, criminal behavior, friends, interpretative phenomenological analysis

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Authors: Jolanta Jonak

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All students demonstrate various learning preferences and learning styles that range from visual, auditory to kinesthetic preferences. These learning preferences are further impacted by individual cognitive profiles hat characterizes itself in linguistic strengths, logical- special, inter-or intra- personal, just to name a few. Students from culturally and linguistically diverse backgrounds (CLD) have an increased risk of being misunderstood by many school systems and even medical personnel. Students with disability, specifically Autism, are faced with another layer of learning differences. Research indicates that large numbers of students are not provided the type of education and types of supports they need in order to be successful in an academic environment. Multiple research findings indicate that significant numbers of school staff self-reports that they do not feel adequately prepared to work with students with disability and different learing profiles. It is very important for the school staff to be educated about different learning needs of students with autism spectrum disorders. Having the knowledge, school staff can avoid unnecessary referrals for office referrals and avoid inaccurate decisions about restrictive learning environments. This presentation will illustrate the cognitive differences in students with autism, how to recognize them, and how to support them through Differentiated Instruction. One way to ensure successful education for students with disability is by providing Differentiated Instruction (DI). DI is quickly gaining its popularity in the Unites States as a scientific- research based instructional approach for all students. This form of support ensures that regardless of the students’ learning preferences and cognitive learning profiles, they have an opportunity to learn through approaches that are suitable to their needs. It is extremely important for the school staff, especially school psychologists who often are the first experts to be consulted by educators, to be educated about differences due to learning preference styles and differentiation needs.

Keywords: special education, autism, differentiation, differences, differentiated instruction

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Authors: Xu Yifei, Le Yining, Yang Qingluan, Tu Yanjie

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Objective: This study aims to investigate the distribution features of Traditional Chinese Medicine (TCM) syndromes and syndrome elements in Epstein-Barr virus-associated diseases and then explores the relations between TCM syndromes or syndrome elements and laboratory indicators of Epstein-Barr virus-associated diseases. Methods: A cross-sectional study of 70 patients with EBV infection was described. We assessed the diagnostic information and laboratory indicators of these patients from Huashan Hospital Affiliated to Fudan University between November 2017 and July 2019. The disease diagnosis and syndrome differentiation were based on the diagnostic criteria of EBV-associated diseases and the theory of TCM respectively. Confidence correlation analysis, logistic regression analysis, cluster analysis, and the Sankey diagram were used to analyze the correlation between the data. Results: The differentiation of the 4 primary TCM syndromes in the collected patients was correlated with the indexes of immune function, liver function, inflammation, and anemia, especially the relationship between Qifen syndrome and high lactic acid dehydrogenase level. The common 11 TCM syndrome elements were associated with the increased CD3+ T cell rate, low hemoglobin level, high procalcitonin level, high lactic acid dehydrogenase level, and low albumin level. Conclusion: The changes in immune function indexes, procalcitonin, and liver function-related indexes in patients with EBV-associated diseases were consistent with the evolution law of TCM syndromes. This study provides a reference for judging the pathological stages of these kinds of diseases, predicting their prognosis, and guiding subsequent treatment strategies based on TCM syndrome type.

Keywords: EBV-associated diseases, traditional Chinese medicine syndrome, syndrome element, diagnostics

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Authors: Laura Victoria Vigoya Morales, David Rolando Suarez Mora

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The problem of detecting enterococcus faecalis is receiving considerable attention with the new cases of beachgoers infected with the bacteria, which can be found in fecal matter. The process detection of this kind of bacteria would be taking a long time, which waste time and money as a result of closing recreation place, like beach or pools. Hence, new methods for automating the process of detecting and recognition of this bacteria has become in a challenge. This article describes a novel approach to detect the enterococcus faecalis bacteria in water by using an octave algorithm, which embody a network neural. This document shows result of performance, quality and integrity of the algorithm.

Keywords: Enterococcus faecalis, image treatment, octave and network neuronal

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Authors: Gi-Hoon Yang, JongHan Ha, MyungGu Yeo, JaeYoon Lee, SeungHyun Ahn, Hyeongjin Lee, HoJun Jeon, YongBok Kim, Minseong Kim, GeunHyung Kim

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Tissue engineering scaffolds must be biocompatible and biodegradable, provide adequate mechanical strength and cell attachment site for proliferation and differentiation. Furthermore, the scaffold morphology (such as pore size, porosity and pore interconnectivity) plays an important role. The electrospinning process has been widely used to fabricate micro/nano-sized fibres. Electrospinning allows for the fabrication of non-woven meshes containing micro- to nano-sized fibers providing high surface-to-volume area for cell attachment. Due to its advantageous characteristics, electrospinning is a useful method for skin, cartilage, bone, and nerve regeneration. In this study, we fabricated PCL scaffolds (SP) consisting of spiral-like struts using 3D melt-plotting system and micro/nanofibers using direct electrospinning writing. By altering the conditions of the conventional melt-plotting method, spiral-like struts were generated. Then, micro/nanofibers were deposited selectively. The control scaffold composed of perpendicular PCL struts was fabricated using the conventional melt-plotting method to compare the cellular activities. The effect on the attached cells (osteoblast-like cells (MG63)) was evaluated depending on the bending instability of the struts. The SP scaffolds showed enhanced biological properties such as initial cell attachment, proliferation and osteogenic differentiation. These results suggest that the SP scaffolds has potential as a bioengineered substitute for soft and hard tissue regeneration.

Keywords: cell attachment, electrospinning, mechanical strength, melt-plotting

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Authors: Yi Li, Yinan Zheng, Ya Ke, Yungwing Ho

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Motor learning is a process that frequently happens among humans and rodents, which is defined as the changes in the capability to perform a skill that is conformed to have a relatively permanent improvement through practice or experience. There are many ways to learn a behavior, among which is observational learning. Observational learning is the process of learning by watching the behaviors of others, for example, a child imitating parents, learning a new sport by watching the training videos or solving puzzles by watching the solutions. Many research explores observational learning in humans and primates. However, the neuronal mechanism of which, especially observational motor learning, was uncertain. It’s well accepted that mirror neurons are essential in the observational learning process. These neurons fire when the primate performs a goal-directed action and sees someone else demonstrating the same action, which suggests they have high firing activity both completing and watching the behavior. The mirror neurons are assumed to mediate imitation or play a critical and fundamental role in action understanding. They are distributed in many brain areas of primates, i.e., posterior parietal cortex (PPC), premotor cortex (M2), and primary motor cortex (M1) of the macaque brain. However, few researchers report the existence of mirror neurons in rodents. To verify the existence of mirror neurons and the possible role in motor learning in rodents, we performed customised string-pulling behavior combined with multiple behavior analysis methods, photometry, electrophysiology recording, c-fos staining and optogenetics in healthy mice. After five days of training, the demonstrator (demo) mice showed a significantly quicker response and shorter time to reach the string; fast, steady and accurate performance to pull down the string; and more precisely grasping the beads. During three days of observation, the mice showed more facial motions when the demo mice performed behaviors. On the first training day, the observer reduced the number of trials to find and pull the string. However, the time to find beads and pull down string were unchanged in the successful attempts on the first day and other training days, which indicated successful action understanding but failed motor learning through observation in mice. After observation, the post-hoc staining revealed that the c-fos expression was increased in the cognitive-related brain areas (medial prefrontal cortex) and motor cortices (M1, M2). In conclusion, this project indicated that the observation led to a better understanding of behaviors and activated the cognitive and motor-related brain areas, which suggested the possible existence of mirror neurons in these brain areas.

Keywords: observation, motor learning, string-pulling behavior, prefrontal cortex, motor cortex, cognitive

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Authors: Juan C. Torrado, Javier Gomez, Guadalupe Montero, German Montoro, M. Dolores Villalba

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One of the most challenging aspects of the executive dysfunction of people with Autism Spectrum Disorders is the behavior control. This is related to a deficit in their ability to regulate, recognize and manage their own emotions. Some researchers have developed applications for tablets and smartphones to practice strategies of relaxation and emotion recognition. However, they cannot be applied to the very moment of temper outbursts, anger episodes or anxiety, since they require to carry the device, start the application and be helped by caretakers. Also, some of these systems are developed for either obsolete technologies (old versions of tablet devices, PDAs, outdated operative systems of smartphones) or specific devices (self-developed or proprietary ones) that create differentiation between the users and the rest of the individuals in their context. For this project we selected smartwatches. Focusing on emergent technologies ensures a wide lifespan of the developed products, because the derived products are intended to be available in the same moment the very technology gets popularized, not later. We also focused our research in commercial versions of smartwatches, since this way differentiation is easily avoided, so the users’ abandonment rate lowers. We have developed a smartwatch system along with a smartphone authoring tool to display self-regulation strategies. These micro-prompting strategies are conformed of pictograms, animations and temporizers, and they are designed by means of the authoring tool: When both devices synchronize their data, the smartwatch holds the self-regulation strategies, which are triggered when the smartwatch sensors detect a remarkable rise of heart rate and movement. The system is being currently tested in an educational center of people with ASD of Madrid, Spain.

Keywords: assistive technologies, emotion regulation, human-computer interaction, smartwatches

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Authors: Danilo López, Nelson Vera, Luis Pedraza

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This paper analyzes fundamental ideas and concepts related to neural networks, which provide the reader a theoretical explanation of Long Short-Term Memory (LSTM) networks operation classified as Deep Learning Systems, and to explicitly present the mathematical development of Backward Pass equations of the LSTM network model. This mathematical modeling associated with software development will provide the necessary tools to develop an intelligent system capable of predicting the behavior of licensed users in wireless cognitive radio networks.

Keywords: neural networks, multilayer perceptron, long short-term memory, recurrent neuronal network, mathematical analysis

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Authors: Aftab Yusuf Raj

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Human milk oligosaccharides (HMOs) are complex carbohydrates. They are 3rd most abundant solid component found in breast milk, after lactose and lipids. HMO has profound beneficial health benefit effects on infants and toddlers. They have diverse roles, in immuno-modulation, development of neonatal gut, influencing the commensal microbiota of developing gut, and anti-inflammatory functions. HMOs, gut and commensal microbiota of the gut work synergistically to bring positive impact on infant and toddler health. HMO influences the gut-brain axis, maintains good mental health and cognitive function and inhibits neuronal inflammation. HMOs are now applied in infant nutrition, and supplementation of HMOs in infant formula is a promising innovation for infant nutrition.

Keywords: HMO’s, gut, epigenetic modulation, bacteria

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Authors: Dana Alina Magdas, Nicoleta Simona Vedeanu, Ioana Feher, Rares Stiufiuc

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Food and beverages authentication using rapid and non-expensive analytical tools represents nowadays an important challenge. In this regard, the potential of vibrational techniques in food authentication has gained an increased attention during the last years. For wines discrimination, Raman spectroscopy appears more feasible to be used as compared with IR (infrared) spectroscopy, because of the relatively weak water bending mode in the vibrational spectroscopy fingerprint range. Despite this, the use of Raman technique in wine discrimination is in an early stage. Taking this into consideration, the wine discrimination potential of surface-enhanced Raman scattering (SERS) technique is reported in the present work. The novelty of this study, compared with the previously reported studies, concerning the application of vibrational techniques in wine discrimination consists in the fact that the present work presents the wines differentiation based on the individual signals obtained from deconvoluted spectra. In order to achieve wines classification with respect to variety, geographical origin and vintage, the peaks intensities obtained after spectra deconvolution were compared using supervised chemometric methods like Linear Discriminant Analysis (LDA). For this purpose, a set of 20 white Romanian wines from different viticultural Romanian regions four varieties, was considered. Chemometric methods applied directly to row SERS experimental spectra proved their efficiency, but discrimination markers identification found to be very difficult due to the overlapped signals as well as for the band shifts. By using this approach, a better general view related to the differences that appear among the wines in terms of compositional differentiation could be reached.

Keywords: chemometry, SERS, variety, wines discrimination

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Authors: Ainur Mukhambetova, Miras Karzhauov, Vyacheslav Ogay

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Introduction: Mesenchymal stem cells (MSCs) have the capacity to be differentiated into several cell lineages and are a promising source for cell therapy and tissue engineering. However, currently most MSCs culturing protocols use media supplemented with fetal bovine serum (FBS), which limits their application in clinic due to the possibility of zoonotic infections, contamination and immunological reactions. Consequently, formulating effective serum free culture medium becomes one of the important problems in contemporary cell biotechnology. Objectives: The aim of this study was to define an optimal serum-free medium for culturing of periosteum derived MSCs. Materials and methods: The MSCs were extracted from human periosteum and transferred to the culture flasks pretreated with CELLstart™. Immunophenotypic characterization, proliferation and in vitro differentiation of cells grown on STEM PRO® MSC SFM were compared to the cells cultured in the standard FBS containing media. Chromosome analysis and flow cytometry were also performed. Results: We have shown that cells were grown on STEM PRO® MSC SFM retained all the morphological, immunophenotypic (CD73, CD90, CD105, vimentin and Stro-1) and cell differentiation characteristics specific to MSCs. Chromosome analysis indicated no anomalies in the chromosome structure. Flow cytometry showed a high expression of cell adhesion molecules CD44 (98,8%), CD90 (97,4%), CD105 (99,1%). In addition, we have shown that cell is grown on STEM PRO® MSC SFM have higher proliferation capacity compared to cell expanded on standard FBS containing the medium. Conclusion: We have shown that STEM PRO® MSC SFM is optimal for culturing periosteum derived human MSCs which subsequently can be safely used in cell therapy.

Keywords: cell technologies, periosteum-derived MSCs, regenerative medicine, serum-free medium

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Authors: Bing–Jing Zhao, Chang-Kui Liu, Hong Wang, Min Hu

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Electron beam rapid manufacturing (EBRM) or Selective laser melting is an additive manufacturing process that uses 3D CAD data as a digital information source and energy in the form of a high-power laser beam or electron beam to create three-dimensional metal parts by fusing fine metallic powders together.Object:The present study was conducted to evaluate the mechanical properties ,the phase transformation,the corrosivity and the biocompatibility of Ti-6Al-4V by EBRM,SLM and forging technique.Method: Ti-6Al-4V alloy standard test pieces were manufactured by EBRM, SLM and forging technique according to AMS4999,GB/T228 and ISO 10993.The mechanical properties were analyzed by universal test machine. The phase transformation was analyzed by X-ray diffraction and scanning electron microscopy. The corrosivity was analyzed by electrochemical method. The biocompatibility was analyzed by co-culturing with mesenchymal stem cell and analyzed by scanning electron microscopy (SEM) and alkaline phosphatase assay (ALP) to evaluate cell adhesion and differentiation, respectively. Results: The mechanical properties, the phase transformation, the corrosivity and the biocompatibility of Ti-6Al-4V by EBRM、SLM were similar to forging and meet the mechanical property requirements of AMS4999 standard. a­phase microstructure for the EBM production contrast to the a’­phase microstructure of the SLM product. Mesenchymal stem cell adhesion and differentiation were well. Conclusion: The property of the Ti-6Al-4V alloy manufactured by EBRM and SLM technique can meet the medical standard from this study. But some further study should be proceeded in order to applying well in clinical practice.

Keywords: 3D printing, Electron Beam Rapid Manufacturing (EBRM), Selective Laser Melting (SLM), Computer Aided Design (CAD)

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Authors: Ahmed Hamid Nassr

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The significant of the old stone ages in the Sudan, seems from the longest times and the large differentiation in antiquities, so many Neolithic cemeteries have been discovered in the Sudan, which the studies revealed costumes’ funerary from the Form of grave and the contents has been contrasted in some elements and constant in the others. Many interpretations of the late Neolithic cemeteries tradition written from the earlier studies, most of them focus of the development and change in the burial from society’s development and ideas of afterlife beginnings. Another emphasis undertaken is the relationship between societies cultural aspects and the environmental conditions of the period; it has been considered that it hyper-affected the development of farming in one way or another. The site of Qalaat Shanan noted in earlier 1990 and excavated by the Author from 2011-2013, the site located north of Khartoum about 170 km in Shendi town. Site setting lies amongst a group of Neolithic sites in the Shendi area, it reflected a succession of various Neolithic activities from the early to the late horizons. Excavation revealed many late Neolithic graves, the adult and child graves show large funerary content, different in quality and quantity, which indicates of societies development and related to religion and ideas about the afterlife. This presentation discusses the variations of the Late Neolithic cemeteries in Galaat Shanan site from the grave form, content and differentiation of the sex and age and compared with other late Neolithic discoveries in Sudan and the Nile valley. The study shows many changes in the graves related to the societies changes in late Neolithic in Sudan, climate and economic subsistence's are well reasons, but also the idea change is the main reason for the changes. That is clear from the variations of grave shape and content in age and sex and might be society’s status and levels.

Keywords: Sudan, Shendi town, Qalaat Shanan, late neolithic, cemeteries, funerary, tradition

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Authors: Karl Kingsley

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Many mechanisms are involved in the control of cellular differentiation and growth, which are often dysregulated in many cancers. Many distinct pathways are involved in these mechanisms of control, including deoxyribonuclease (DNA) methyltransferase and histone deacetylase (HDAC) activation that controls both genetic and epigenetic modifications and micro ribonucleic acid (RNA) expression. Less is known about the expression of DNA methyltransferase (DNMT) and HDAC in oral cancers and the effect on microRNA expression. The primary objective of this study was to evaluate the expression of DNMT and HDAC family members in oral cancer and the concomitant expression of cancer-associated microRNAs. Using commercially available oral cancers, including squamous cell carcinoma (SCC)-4, SCC-9, SCC-15, and SCC-25, RNA was extracted and screened for DNMT, HDAC, and microRNA expression using highly-specific primers and quantitative polymerase chain reaction (qPCR). These data revealed low or absent expression of DNMT-1, which is associated with cellular differentiation but increased expression of DNMT-3a and DNMT-3b in all SCC cell lines compared with normal non-cancerous cell controls. In addition, no expression of HDAC1 and HDAC2 expression was found among the normal, non-cancerous cells but was highly expressed in each of the SCC cell lines examined. Differential expression of oncogenic and cancer-associated microRNAs was also observed among the SCC cell lines, including miR-21, miR-133, miR-149, miR-155, miR-365, and miR-720. These findings also appeared to vary according to observed growth rates among these cells. These data may be the first to demonstrate the expression and association between HDAC and DNMT3 family members among oral cancers. In addition, the differential expression of these epigenetic modifiers may be associated with the expression of specific microRNAs in these cancers, which have not previously been observed to the best of the author's knowledge. In addition, some associations and relationships may exist between the expression of these biomarkers and the rates of growth and proliferation, which may suggest that these expression patterns might represent potentially useful biomarkers to determine tumor aggressiveness and other phenotypic behaviors among oral cancers.

Keywords: oral cancer, DNA methyltransferase, histone deacetylase, microRNA

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Authors: M. Nerantzaki, I. Koliakou, D. Bikiaris

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This study evaluates the hypothesis that the incorporation of fibrous hydroxyapatite nanoparticles (nHA) with high crystallinity and high aspect ratio, synthesized by hydrothermal method, into Poly(butylene succinate) (PBSu), improves the bioactivity of the aliphatic polyester and affects new bone growth inhibiting resorption and enhancing bone formation. Hydroxyapatite nanorods were synthesized using a simple hydrothermal procedure. First, the HPO42- -containing solution was added drop-wise into the Ca2+-containing solution, while the molar ratio of Ca/P was adjusted at 1.67. The HA precursor was then treated hydrothermally at 200°C for 72 h. The resulting powder was characterized using XRD, FT-IR, TEM, and EDXA. Afterwards, PBSu nanocomposites containing 2.5wt% (nHA) were prepared by in situ polymerization technique for the first time and were examined as potential scaffolds for bone engineering applications. For comparison purposes composites containing either 2.5wt% micro-Bioglass (mBG) or 2.5wt% mBG-nHA were prepared and studied, too. The composite scaffolds were characterized using SEM, FTIR, and XRD. Mechanical testing (Instron 3344) and Contact Angle measurements were also carried out. Enzymatic degradation was studied in an aqueous solution containing a mixture of R. Oryzae and P. Cepacia lipases at 37°C and pH=7.2. In vitro biomineralization test was performed by immersing all samples in simulated body fluid (SBF) for 21 days. Biocompatibility was assessed using rat Adipose Stem Cells (rASCs), genetically modified by nucleofection with DNA encoding SB100x transposase and pT2-Venus-neo transposon expression plasmids in order to attain fluorescence images. Cell proliferation and viability of cells on the scaffolds were evaluated using fluoresce microscopy and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay. Finally, osteogenic differentiation was assessed by staining rASCs with alizarine red using cetylpyridinium chloride (CPC) method. TEM image of the fibrous HAp nanoparticles, synthesized in the present study clearly showed the fibrous morphology of the synthesized powder. The addition of nHA decreased significantly the contact angle of the samples, indicating that the materials become more hydrophilic and hence they absorb more water and subsequently degrade more rapidly. In vitro biomineralization test confirmed that all samples were bioactive as mineral deposits were detected by X-ray diffractometry after incubation in SBF. Metabolic activity of rASCs on all PBSu composites was high and increased from day 1 of culture to day 14. On day 28 metabolic activity of rASCs cultured on samples enriched with bioceramics was significantly decreased due to possible differentiation of rASCs to osteoblasts. Staining rASCs with alizarin red after 28 days in culture confirmed our initial hypothesis as the presence of calcium was detected, suggesting osteogenic differentiation of rACS on PBSu/nHAp/mBG 2.5% and PBSu/mBG 2.5% composite scaffolds.

Keywords: biomaterials, hydroxyapatite nanorods, poly(butylene succinate), scaffolds

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Authors: Iliyasu Musa Omoyine, Musa Sunday Abraham, Oladele Sunday Blessing, Iliya Ibrahim Abdullahi, Ibegbu Augustine Oseloka, Nuhu Nana-Hawau, Animoku Abdulrazaq Amoto, Yusuf Abdullateef Onoruoiza, Sambo Sohnap James, Akpulu Steven Peter, Ajayi Abayomi

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The neuroprotective role of inflammation from detrimental intrinsic and extrinsic factors has been reported. However, the overactivation of astrocytes and microglia due to lead toxicity produce excessive pro-inflammatory cytokines, mediating neurodegenerative diseases. The present study investigated the mitigatory effects of quercetin on neuroinflammation, correlating with memory function in lead-exposed rats. In this study, Wistar rats were administered orally with Quercetin (Q: 60 mg/kg) and Succimer as a standard drug (S: 10 mg/kg) for 21 days after lead exposure (Pb: 125 mg/kg) of 21 days or in combination with Pb, once daily for 42 days. Working and reference memory was assessed using an Eight-arm radial water maze (8-ARWM). The changes in brain lead level, the neuronal nitric oxide synthase (nNOS) activity, and the level of neuroinflammatory markers such as tumour necrosis factor-alpha (TNF-α) and Interleukin 1 Beta (IL-1β) were determined. Immunohistochemically, astrocyte expression was evaluated. The results showed that the brain level of lead was increased significantly in lead-exposed rats. The expression of astrocytes increased in the CA3 and CA1 regions of the hippocampus, and the levels of brain TNF-α and IL-1β increased in lead-exposed rats. Lead impaired reference and working memory by increasing reference memory errors and working memory incorrect errors in lead-exposed rats. However, quercetin treatment effectively improved memory and inhibited neuroinflammation by reducing astrocytes’ expression and the levels of TNF-α and IL-1β. The expression of astrocytes and the levels of TNF-α and IL-1β correlated with memory function. The possible explanation for quercetin’s anti-neuroinflammatory effect is that it modulates the activity of cellular proteins involved in the inflammatory response; inhibits the transcription factor of nuclear factor-kappa B (NF-κB), which regulates the expression of proinflammatory molecules; inhibits kinases required for the synthesis of Glial fibrillary acidic protein (GFAP) and modifies the phosphorylation of some proteins, which affect the structure and function of intermediate filament proteins; and, lastly, induces Cyclic-AMP Response Element Binding (CREB) activation and neurogenesis as a compensatory mechanism for memory deficits and neuronal cell death. In conclusion, the levels of neuroinflammatory markers negatively correlated with memory function. Thus, quercetin may be a promising therapy in neuroinflammation and memory dysfunction in populations prone to lead exposure.

Keywords: lead, quercetin, neuroinflammation, memory

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Authors: Vera Sovkova, Andrea Mickova, Matej Buzgo, Karolina Vocetkova, Eva Filova, Evzen Amler

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PCL (poly-ε-caprolactone) nanofibrous scaffolds with adhered liposomes were prepared and tested as a possible drug delivery system for various synthetic growth factors. TGFβ, bFGF, and IGF-I have been shown to increase hMSC (human mesenchymal stem cells) proliferation and to induce hMSC differentiation. Functionalized PCL nanofibers were prepared with synthetic growth factors encapsulated in liposomes adhered to them in three different concentrations. Other samples contained PCL nanofibers with adhered, free synthetic growth factors. The synthetic growth factors free medium served as a control. The interaction of liposomes with the PCL nanofibers was visualized by SEM, and the release kinetics were determined by ELISA testing. The potential of liposomes, immobilized on the biodegradable scaffolds, as a delivery system for synthetic growth factors, and as a suitable system for MSCs adhesion, proliferation and differentiation in vitro was evaluated by MTS assay, dsDNA amount determination, confocal microscopy, flow cytometry and real-time PCR. The results showed that the growth factors adhered to the PCL nanofibers stimulated cell proliferation mainly up to day 11 and that subsequently their effect was lower. By contrast, the release of the lowest concentration of growth factors from liposomes resulted in gradual proliferation of MSCs throughout the experiment. Moreover, liposomes, as well as free growth factors, stimulated type II collagen production, which was confirmed by immunohistochemical staining using monoclonal antibody against type II collagen. The results of this study indicate that growth factors enriched liposomes adhered to surface of PCL nanofibers could be useful as a drug delivery instrument for application in short timescales, be combined with nanofiber scaffolds to promote local and persistent delivery while mimicking the local microenvironment. This work was supported by project LO1508 from the Ministry of Education, Youth and Sports of the Czech Republic

Keywords: drug delivery, growth factors, hMSC, liposomes, nanofibres

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Authors: M. Chojak

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Recently, more and more scientific disciplines refer to research in the field of neurobiology. Interdisciplinary research procedures are created using modern methods of brain imaging. Neither did the pedagogues start looking for neuronal conditions for various processes. The publications began to show concepts such as ‘neuropedagogy’, ‘neuroeducation’, ‘neurodidactics’, ‘brain-friendly education’. They were and are still used interchangeably. In the offer of training for teachers, the topics of multiple intelligences or educational kinesiology began to be more and more popular. These and other ideas have been actively introduced into the curricula. To our best knowledge, the literature on the subject lacks articles organizing the new nomenclature and indicating the methodological framework for research that would confirm the effectiveness of the above-mentioned innovations. The author of this article tries to find the place for neuropedagogy in the system of sciences, define its subject of research, methodological framework and basic concepts. This is necessary to plan studies that will verify the so-called neuromyths.

Keywords: brain, education, neuropedagogy, research

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Authors: Wei-Wen Hu, Yong-Zhi Zhong

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Depositing conductive polypyrrole (PPy) onto elastic polydimethylsiloxane (PDMS) substrate can obtain a highly stretchable conductive film, which can be used to construct a bioreactor to cyclically stretch and electrically stimulate surface cells. However, how to completely harvest these stimulated muscle tissue to repair damaged muscle is a challenge. To address this concern, N-isopropylacrylamide (NIPAAm), a monomer of temperature-sensitive polymer, was added during the polymerization of pyrrole on PDMS so that the resulting P(Py-co-NIPAAm)/PDMS should own both conductivity and thermo-sensitivity. Therefore, cells after stimulation can be completely harvested as cell sheets by reducing temperature. Mouse skeletal myoblast, C2C12 cells, were applied to examine our hypothesis. In electrical stimulation, C2C12 cells on P(Py-co-NIPAAm)/PDMS demonstrated the best myo-differentiation under the electric field of 1 V/cm. Regarding cyclic stretching, the strain equal to or higher than 9% can highly align C2C12 perpendicular to the stretching direction. The Western blotting experiments demonstrated that the cell sheets harvested by cooling reserved more extracellular matrix (ECM) than cells collected by the traditional trypsin digestion method. Immunostaining of myosin heavy chain protein (MHC) indicated that both mechanical and electrical stimuli effectively increased the number of myotubes and the differentiation ratio, and the myotubes can be aligned by cyclic stretching. Stimulated cell sheets can be harvested by cooling, and the alignment of myotubes was still maintained. These results suggested that the deposition of P(Py-co-NIPAAm) on PDMS can be applied to harvest intact cell sheets after cyclic stretching and electrical stimulation, which increased the feasibility of bioreactor for the application of tissue engineering and regenerative medicine.

Keywords: bioreactor, cell sheet, conductive polymer, cyclic stretching, electrical stimulation, muscle tissue engineering, myogenesis, thermosensitive hydrophobicity

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Authors: Huseyin Apdik, Aysegul Dogan, Selami Demirci, Ezgi Avsar Apdik, Fikrettin Sahin

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Mesenchymal stem cells (MSCs) are crucial cell types for bone maintenance and growth along with resident bone progenitor cells providing bone tissue integrity during osteogenesis and skeletal growth. Any deficiency in this regulation would result in vital bone diseases. Of those, osteoporosis, characterized by a reduction in bone mass and mineral density, is a critical skeletal disease for especially elderly people. The commonly used drugs for the osteoporosis treatment are bisphosphonates (BPs). The most prominent role of BPs is to prevent bone resorption arisen from high osteoclast activity. However, administrations of bisphosphonates may also cause bisphosphonate-induced osteonecrosis of the jaw (BIONJ). Up to the present, the researchers have proposed several circumstances for BIONJ. However, effects of long-term and/or high dose usage of BPs on stem cell’s proliferation, survival, differentiation or maintenance capacity have not been evaluated yet. The present study will be held to; figure out BPs’ effects on MSCs in vitro in the aspect of cell proliferation and toxicity, migration, angiogenic activity, lineage specific gene and protein expression levels, mesenchymal stem cell properties and potential signaling pathways affected by BP treatment. Firstly, mesenchymal stem cell characteristics of Dental Pulp Stem Cells (DPSCs) and Periodontal Ligament Stem Cells (PDLSCs) were proved using flow cytometry analysis. Cell viability analysis was completed to determine the cytotoxic effects of BPs (Zoledronate (Zol), Alendronate (Ale) and Risedronate (Ris)) on DPSCs and PDLSCs by the 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS) assay. Non-toxic concentrations of BPs were determined at 24 h under growth condition, and at 21 days under osteogenic differentiation condition for both cells. The scratch assay was performed to evaluate their migration capacity under the usage of determined of BPs concentrations at 24 h. The results revealed that while the scratch closure is 70% in the control group for DPSCs, it was 57%, 66% and 66% in Zol, Ale and Ris groups, respectively. For PDLSs, while wound closure is 71% in control group, it was 65%, 66% and 66% in Zol, Ale and Ris groups, respectively. As future experiments, tube formation assay and aortic ring assay will be done to determinate angiogenesis abilities of DPSCs and PDLSCs treated with BPs. Expression levels of osteogenic differentiation marker genes involved in bone development will be determined using real time-polymerase change reaction (RT-PCR) assay and expression profiles of important proteins involved in osteogenesis will be evaluated using western blotting assay for osteogenically differentiated MSCs treated with or without BPs. In addition to these, von Kossa staining will be performed to measure calcium mineralization status of MSCs.

Keywords: bisphosphonates, bisphosphonate-induced osteonecrosis of the jaw, mesenchymal stem cells, osteogenesis

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