Effect of oxidative stress upon absorption of glucose by the human placenta: in vitro studies with BeWo cells

Pregnancy is a dynamic state and the placenta is a temporary organ that, among other important functions, plays a crucial role in the transport of nutrients and metabolites between the mother and the fetus, which is essential for a successful pregnancy. Among these nutrients, glucose is considered a primary source of energy and, therefore, fundamental to insure proper fetus development. Several studies have shown that glucose uptake is dependent on several morphological and biochemical placental conditions. Oxidative stress results from the unbalance between reactive oxygen species (ROS) and antioxidants, in favor of the first. During pregnancy, ROS, and therefore oxidative stress, increase, due to increased tissue oxygenation. Moreover, the relation between ROS and some pathological conditions during pregnancy has been well established. For these reasons, it becomes essential to understand if oxidative stress can compromise the uptake of glucose by the placenta. To make this study possible, a trophoblastic cell line, the BeWo cell line, was used. Experiments regarding glucose uptake, either under normal or oxidative stress conditions, were conducted using tert-butylhydroperoxide (tBOOH) as an oxidative stress inducer, and 3H-2-deoxy-D-glucose (3H-DG) as a glucose analogue. Afterwards, studies regarding the involvement of glucose facilitative transporters (GLUT) and the phosphatidylinositol 3-kinases (PI3K) and protein kinase C (PKC) pathways were conducted, also under normal and oxidative stress conditions. A few antioxidants, endogenous and from diet, were also tested in order to study their possible reversible effect of the oxidative effect of tBOOH upon apical 3H-DG uptake. Finally, transepithelial studies gave interesting insights regarding the apical-to-basolateral transport of 3H-DG. Results showed that 3H-DG uptake, in BeWo cells, is roughly 50% GLUT-mediated and that tBOOH (100 μM; 24h) decreases apical 3H-DG uptake in BeWo cells by about 33%, by reducing both GLUT- (by 28%) and non-GLUT-mediated (by 40%) 3H-DG uptake. Uptake of 3H-DG and the effect of tBOOH upon 3H-DG uptake are not dependent on PKC and PI3K. Moreover, the effect of tBOOH is not associated with a reduction in GLUT1 mRNA levels. Resveratrol, quercetin and epigallocatechin-3-gallate, at 50 μM, reversed, by at least 45%, the effect of tBOOH upon 3H-DG uptake. Transwell studies show that the apical-to-basolateral transepithelial transport of 3H-DG is increased by tBOOH.In conclusion, our results show that tBOOH caused a marked decrease in both GLUT and non-GLUT-mediated apical uptake of 3H-DG by BeWo cells. Given the association of increased oxidative stress levels with several important pregnancy pathologies, and the important role of glucose for fetal development, the results of this study appear very interesting.

Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells

Bone is constantly being molded and shaped by the action of osteoclasts and osteoblasts. A proper equilibrium between both cell types metabolic activities is required to ensure an adequate skeletal tissue structure, and it involves resorption of old bone and formation of new bone tissue. It is reported that treatment with antiepileptic drugs (AEDs) can elicit alterations in skeletal structure, in particular in bone mineral density. Nevertheless, the knowledge regarding the effects of AEDs on bone cells are still scarce. In this context, the aim of this study was to investigate the effects of five different AEDs on human osteoclastic, osteoblastic and co-cultured cells. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood and were characterized for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. Osteoblastic cell cultures were obtained from femur heads of patients (25-45 years old) undergoing orthopaedic surgery procedures and were then studied for cellular proliferation/viability, ALP activity, histochemical staining of ALP and apoptosis rate. Also the expression of osteoblast-related genes and the involvement of some osteoblastogenesis-related signalling pathways on cellular response were addressed. For co-cultured cells, osteoblastic cells were firstly seeded and cultured. After that, PBMC were added to the osteoblastic cells and co-cultures were evaluated using the same osteoclast and osteoblast parameters mentioned above for the corresponding isolated cell. Cell-cultures were maintained in the absence (control) or in the presence of different AEDs (carbamazepine, gabapentin, lamotrigine, topiramate and valproic acid). All the tested drugs were able to affect osteoclastic and osteoblastic cells development, although with different profiles on their osteoclastogenic and osteoblastogenic modulation properties. Globally, the tendency was to inhibit the process. Furthermore, the signaling pathways involved in the process also seemed to be differently affected by the AEDs, suggesting that the different drugs may affect osteoclastogenesis and/or osteoblastogenesis through different mechanisms. In conclusion, the present study showed that the different AEDs had the ability to directly and indirectly modulate bone cells differentiation, shedding new light towards a better understanding of how these drugs can affect bone tissue.

Eastwards / westwards: which direction for gender studies in the 21st century?

Eastwards / Westwards: Which Direction for Gender Studies in the XXIst Century? is a collection of essays which focus on themes and methods that characterize current research into gender in Asian countries in general. In this collection, ideas derived from Gender Studies elsewhere in the world have been subjected to scrutiny for their utility in helping to describe and understand regional phenomena. But the concepts of Local and Global – with their discoursive productions – have not functioned as a binary opposition: localism and globalism are mutually constitutive and researchers have interrogated those spaces of interaction between the ‘self’ and the ‘other’, bearing in mind their own embeddedness in social and cultural structures and their own historical memory. Contributors to this collection provided a critical transnational perspective on some of the complex effects of the dynamics of cultural globalization, by exploring the relation between gender and development, language, historiography, education and culture. We have also given attention to the ideological and rhetorical processes through which gender identity is constructed, by comparing textual grids and patterns of expectation. Likewise, we have discussed the role of ethnography, anthropology, historiography, sociology, fiction, popular culture and colonial and post-colonial sources in (re)inventing old/new male/female identities, their conversion into concepts and circulation through time and space. This multicultural and trans-disciplinary selection of essays is totally written in English, fully edited and revised, therefore, it has a good potential for an immediate international circulation. This project may trace new paths and issues for discussion on what concerns the life, practices and narratives by and about women in Asia, as well as elsewhere in the present day global experience. Academic readership: Researchers, scholars, educators, graduate and post-graduate students, doctoral students and general non-fiction readers, with a special interest in Gender Studies, Asia, Colonial and Post-Colonial Literature, Anthropology, Cultural Studies, History, Historiography, Politics, Race, Feminism, Language, Linguistics, Power, Political and Feminist Agendas, Popular Culture, Education, Women’s Writing, Religion, Multiculturalism, Globalisation, Migration. Chapter summary: 1. “Social Gender Stereotypes and their Implication in Hindi”, Anjali Pande, Jawaharlal Nehru University, New Delhi, India. This essay looks at the subtle ways in which gender identities are constructed and reinforced in India through social norms of language use. Language itself becomes a medium for perpetuating gender stereotypes, forcing its speakers to confirm to socially defined gender roles. Using examples from a classroom discussion about a film, this essay will highlight the underlying rigid male-female stereotypes in Indian society with their more obvious expressions in language. For the urban woman in India globalisation meant increased economic equality and exposure to changed lifestyles. On an individual level it also meant redefining gender relations and changing the hierarchy in man-­woman relationships. With the economic independence there is a heightened sense of liberation in all spheres of social life, a confidence to fuzz the rigid boundaries of gender roles. With the new films and media celebrating this liberated woman, who is ready to assert her sexual needs, who is ready to explode those long held notions of morality, one would expect that the changes are not just superficial. But as it soon became obvious in the course of a classroom discussion about relationships and stereotypes related to age, the surface changes can not become part of the common vocabulary, for the obvious reason that there is still a vast gap between the screen image of this new woman and the ground reality. Social considerations define the limits of this assertiveness of women, whereas men are happy to be liberal within the larger frame of social sanctions. The educated urban woman in India speaks in favour of change and the educated urban male supports her, but one just needs to scratch the surface to see the time tested formulae of gender roles firmly in place. The way the urban woman happily balances this emerging promise of independence with her gendered social identity, makes it necessary to rethink some aspects of looking at gender in a gradually changing, traditional society like India. 2. “The Linguistic Dimension of Gender Equality”, Alissa Tolstokorova, Kiev Centre for Gender Information and Education, Ukraine. The subject-matter of this essay is gender justice in language which, as I argue, may be achieved through the development of a gender-related approach to linguistic human rights. The last decades of the 20th century, globally marked by a “gender shift” in attitudes to language policy, gave impetus to the social movement for promoting linguistic gender equality. It was initiated in Western Europe and nowadays is moving eastwards, as ideas of gender democracy progress into developing countries. But, while in western societies gender discrimination through language, or linguistic sexism, was an issue of concern for over three decades, in developing countries efforts to promote gender justice in language are only in their infancy. My argument is that to promote gender justice in language internationally it is necessary to acknowledge the rights of women and men to equal representation of their gender in language and speech and, therefore, raise a question of linguistic rights of the sexes. My understanding is that the adoption of the Universal Declaration of Linguistic Rights in 1996 provided this opportunity to address the problem of gender justice in language as a human rights issue, specifically as a gender dimension of linguistic human rights. 3. “The Rebirth of an Old Language: Issues of Gender Equality in Kazakhstan”, Maria Helena Guimarães, Polytechnic Institute of Porto, Portugal. The existing language situation in Kazakhstan, while peaceful, is not without some tension. We propose to analyze here some questions we consider relevant in the frame of cultural globalization and gender equality, such as: free from Russian imperialism, could Kazakhstan become an easy prey of Turkey’s “imperialist dream”? Could these traditionally Muslim people be soon facing the end of religious tolerance and gender equality, becoming this new old language an easy instrument for the infiltration in the country of fundamentalism (it has already crossed the boarders of Uzbekistan), leading to a gradual deterioration of its rich multicultural relations? The present structure of the language is still very fragile: there are three main dialects and many academics defend the re-introduction of the Latin alphabet, thus enlarging the possibility of cultural “contamination” by making the transmission of fundamentalist ideas still easier through neighbour countries like Azerbaijan, Uzbekistan and Turkmenistan (their languages belong to the same sub-group of Common Turkic), where the Latin alphabet is already in use, and where the ground for such ideas shown itself very fruitful. 4. “Construction of Womanhood in the Bengali Language of Bangladesh”, Raasheed Mahmood; University of New South Wales, Sydney. The present essay attempts to explore the role of gender-based language differences and of certain markers that reveal the status accorded to women in Bangladesh. Discrimination against women, in its various forms, is endemic in communities and countries around the world, cutting across class, race, age, and religious and national boundaries. One cannot understand the problems of gender discrimination solely by referring to the relationship of power or authority between men and women. Rather one needs to consider the problem by relating it to the specific social formation in which the image of masculinity and femininity is constructed and reconstructed. Following such line of reasoning this essay will examine the nature of gender bias in the Bengali language of Bangladesh, holding the conviction that as a product of social reality language reflects the socio-cultural behaviour of the community who speaks it. This essay will also attempt to shed some light on the processes through which gender based language differences produce actual consequences for women, who become exposed to low self-esteem, depression and systematic exclusion from public discourse. 5. “Marriage in China as an expression of a changing society”, Elisabetta Rosado David, University of Porto, Portugal, and Università Ca’Foscari, Venezia, Italy. In 29 April 2001, the new Marriage Law was promulgated in China. The first law on marriage was proclaimed in 1950 with the objective of freeing women from the feudal matrimonial system. With the second law, in 1981, values and conditions that had been distorted by the Cultural Revolution were recovered. Twenty years later, a new reform was started, intending to update marriage in the view of the social and cultural changes that occurred with Deng Xiaoping’s “open policy”. But the legal reform is only the starting point for this case-study. The rituals that are followed in the wedding ceremony are often hard to understand and very difficult to standardize, especially because China is a vast country, densely populated and characterized by several ethnic minorities. Two key words emerge from this issue: syncretism and continuity. On this basis, we can understand tradition in a better way, and analyse whether or not marriage, as every social manifestation, has evolved in harmony with Chinese culture. 6. “The Other Woman in the Portuguese Colonial Empire: The Case of Portuguese India”, Maria de Deus Manso, University of Évora, Portugal. This essay researches the social, cultural and symbolic history of local women in the Portuguese Indian colonial enclaves. The normative Portuguese overseas history has not paid any attention to the “indigenous” female populations in colonial Portuguese territories, albeit the large social importance of these social segments largely used in matrimonial and even catholic missionary strategies. The first attempt to open fresh windows in the history of this new field was the publication of Charles Boxer’s referential study about Women in lberian Overseas Expansion, edited in Portugal only after the Revolution of 1975. After this research we can only quote some other fragmentary efforts. In fact, research about the social, cultural, religious, political and symbolic situation of women in the Portuguese colonial territories, from the XVI to the XX century, is still a minor historiographic field. In this essay we discuss this problem and we study colonial representations of women in the Portuguese Indian enclaves, mainly in the territory of Goa, using case studies methodologies. 7. “Heading East this Time: Critical Readings on Gender in Southeast Asia”, Clara Sarmento, Polytechnic Institute of Porto, Portugal. This essay intends to discuss some critical readings of fictional and theoretical texts on gender condition in Southeast Asian countries. Nowadays, many texts about women in Southeast Asia apply concepts of power in unusual areas. Traditional forms of gender hegemony have been replaced by other powerful, if somewhat more covert, forms. We will discuss some universal values concerning conventional female roles as well as the strategies used to recognize women in political fields traditionally characterized by male dominance. Female empowerment will mean different things at different times in history, as a result of culture, local geography and individual circumstances. Empowerment needs to be perceived as an individual attitude, but it also has to be facilitated at the macro­level by society and the State. Gender is very much at the heart of all these dynamics, strongly related to specificities of historical, cultural, ethnic and class situatedness, requiring an interdisciplinary transnational approach.

Evaluation of 99mTc-Sestamibi as a potential tool to investigate PgP activity in inflammation

Introduction: In the XXI Century ’s Society the scientific investigation process has been growing steadily , and the field of the pharmaceutical research is one of the most enthusiastic and relevant . Here, it is very important to correlate observed functional alterations with possibly modified drug bio distribution patterns . Cancer, inflammation and inf ection are processes that induce many molecular intermediates like cytokines, chemokines and other chemical complexes that can alter the pharmacokinetics of many drugs. One cause of such changes is thought to be the modulator action of these complexes in t he P - Glyco p rotein activity, because they can act like inducers/inhibitors of MDR - 1 expression. This protein results from the expression of MDR - 1 gene, and acts as an ATP energy - dependent efflux pump, with their substrates including many drugs , like antiretrovirals, anticancers, anti - infectives, immunosuppressants, steroids or opioids . Objectives: Because of the lack of methods to provide helpful information in the investigation of in vivo molecular changes in Pgp activity during infection/infl ammation processes, and its value in the explanation of the altered drug pharmacokinetic, this paper want to evaluate the potential utility of 99m Tc - Sestamibi scintigraphy during this kind of health sciences investigation. Although the a im is indeed to create a technique to the in vivo study of Pgp activity, this preliminary Project only reaches the in vitro study phase, assumed as the first step in a n evaluation period for a new tool development. Materials and Methods: For that reason , we are performing in vitro studies of influx and efflux of 99m Tc - Sestamibi ( that is a substrate of Pgp) in hepatocytes cell line (HepG2). We are interested in clarify the cellular behavior of this radiopharmaceutical in Lipopolysaccharide(LPS) stimulated cells ( well known in vitro model of inflammation) to possibly approve this methodology. To validate the results, the Pgp expression will be finally evaluated using Western Blot technique. Results: Up to this moment , we still don’t have the final results, but we have already enough data to let us believe that LPS stimulation induce a downregulation of MDR - 1, and consequently Pgp, which could conduce to a prolonged retention of 99m Tc - Sestamibi in the inflamed cells . Conclusions: If and when this methodology demonstrate the promising results we expect, one will be able to con clude that Nuclear Medicine is an important tool to help evidence based research also on this specific field .

Macrophages as biomarkers in BCG treatment response in bladder cancer

Bladder cancer is a common urologic cancer and the majority has origin in the urothelium. Patients with intermediate and high risk of recurrence/progression bladder cancer are treated with intravesical instillation with Bacillus Calmette-Guérin, however, approximately 30% of patients do not respond to treatment. At the moment, there are no accepted biomarkers do predict treatment outcome and an early identification of patients better served by alternative therapeutics. The treatment initiates a cascade of cytokines responsible by recruiting macrophages to the tumor site that have been shown to influence treatment outcome. Effective BCG therapy needs precise activation of the Th1 immune pathway associated with M1 polarized macrophages. However, tumor-associated macrophages (TAMs) often assume an immunoregulatory M2 phenotype, either immunosuppressive or angiogenic, that interfere in different ways with the BCG induced antitumor immune response. The M2 macrophage is influenced by different microenvironments in the stroma and the tumor. In particular, the degree of hypoxia in the tumors is responsible by the recruitment and differentiation of macrophages into the M2 angiogenic phenotype, suggested to be associated with the response to treatment. Nevertheless, neither the macrophage phenotypes present nor the influence of localization and hypoxia have been addressed in previous studies. Therefore, this work devoted to study the influence of TAMs, in particular of the M2 phenotype taking into account their localization (stroma or tumor) and the degree of hypoxia in the tumor (low or high) in BCG treatment outcome. The study included 99 bladder cancer patients treated with BCG. Tumors resected prior to treatment were evaluated using immunohistochemistry for CD68 and CD163 antigens, which identify a lineage macrophage marker and a M2-polarized specific cell surface receptor, respectively. Tumor hypoxia was evaluated based on HIF-1α expression. As a main finding it was observed that a high predominance of CD163+ macrophage counts in the stroma of tumors under low hypoxia was associated with BCG immunotherapy failure, possibly due to its immunosuppressive phenotype. This study further reinforces the importance the tumor microenvironment in the modulation of BCG responses.

Anti-tumoral effect of the non-nucleoside DNMT inhibitor RG108 in human prostate cancer cells

Background: Current therapeutic strategies for advanced prostate cancer (PCa) are largely ineffective. Because aberrant DNA methylation associated with inappropriate gene-silencing is a common feature of PCa, DNA methylation inhibitors might constitute an alternative therapy. In this study we aimed to evaluate the anti-cancer properties of RG108, a novel non-nucleoside inhibitor of DNA methyltransferases (DNMT), in PCa cell lines. Methods: The anti-tumoral impact of RG108 in LNCaP, 22Rv1, DU145 and PC-3 cell lines was assessed through standard cell viability, apoptosis and cell cycle assays. Likewise, DNMT activity, DNMT1 expression and global levels of DNA methylation were evaluated in the same cell lines. The effectiveness of DNA demethylation was further assessed through the determination of promoter methylation and transcript levels of GSTP1, APC and RAR-β2, by quantitative methylation-specific PCR and RT-PCR, respectively. Results: RG108 led to a significant dose and time dependent growth inhibition and apoptosis induction in LNCaP, 22Rv1 and DU145. LNCaP and 22Rv1 also displayed decreased DNMT activity, DNMT1 expression and global DNA methylation. Interestingly, chronic treatment with RG108 significantly decreased GSTP1, APC and RAR-β2 promoter hypermethylation levels, although mRNA re-expression was only attained GSTP1 and APC. Conclusions: RG108 is an effective tumor growth suppressor in most PCa cell lines tested. This effect is likely mediated by reversion of aberrant DNA methylation affecting cancer related-genes epigenetically silenced in PCa. However, additional mechanism might underlie the anti-tumor effects of RG108. In vivo studies are now mandatory to confirm these promising results and evaluate the potential of this compound for PCa therapy.

Efficient computation of min and max sensor values in multihop networks

Consider a wireless sensor network (WSN) where a broadcast from a sensor node does not reach all sensor nodes in the network; such networks are often called multihop networks. Sensor nodes take individual sensor readings, however, in many cases, it is relevant to compute aggregated quantities of these readings. In fact, the minimum and maximum of all sensor readings at an instant are often interesting because they indicate abnormal behavior, for example if the maximum temperature is very high then it may be that a fire has broken out. In this context, we propose an algorithm for computing the min or max of sensor readings in a multihop network. This algorithm has the particularly interesting property of having a time complexity that does not depend on the number of sensor nodes; only the network diameter and the range of the value domain of sensor readings matter.

Highly scalable aggregate computations in cyber-physical systems: physical environment meets communication protocols

In this paper, we focus on large-scale and dense Cyber- Physical Systems, and discuss methods that tightly integrate communication and computing with the underlying physical environment. We present Physical Dynamic Priority Dominance ((PD)2) protocol that exemplifies a key mechanism to devise low time-complexity communication protocols for large-scale networked sensor systems. We show that using this mechanism, one can compute aggregate quantities such as the maximum or minimum of sensor readings in a time-complexity that is equivalent to essentially one message exchange. We also illustrate the use of this mechanism in a more complex task of computing the interpolation of smooth as well as non-smooth sensor data in very low timecomplexity.

A fractional calculus perspective in the evolutionary design of combinational circuits

This paper analyses the performance of a genetic algorithm (GA) in the synthesis of digital circuits using two novel approaches. The first concept consists in improving the static fitness function by including a discontinuity evaluation. The measure of variability in the error of the Boolean table has similarities with the function continuity issue in classical calculus. The second concept extends the static fitness by introducing a fractional-order dynamical evaluation.

Students’ reading behavior: digital vs. print preferences in portuguese context

The aim of this paper is to present the main Portuguese results from a multi-national study on reading format preferences and behaviors from undergraduate students from Polytechnic Institute of Porto (Portugal). For this purpose we apply an adaptation of the Academic Reading Questionnaire previously created by Mizrachi (2014). This survey instrument has 14 Likert-style statements regarding the format influence in the students reading behavior, including aspects such as ability to remember, feelings about access convenience, active engagement with the text by highlighting and annotating, and ability to review and concentrate on the text. The importance of the language and dimension of the text to determine the preference format is also inquired. Students are also asked about the electronic device they use to read digital documents. Finally, some demographic and academic data were gathered. The analysis of the results will be contextualized on a review of the literature concerning youngsters reading format preferences. The format (digital or print) in which a text is displayed and read can impact comprehension, which is an important information literacy skill. This is a quite relevant issue for class readings in academic context because it impacts learning. On the other hand, students preferences on reading formats will influence the use of library services. However, literature is not unanimous on this subject. Woody, Daniel and Baker (2010) concluded that the experience of reading is not the same in electronic or print context and that students prefer print books than e-books. This thesis is reinforced by Ji, Michaels and Waterman (2014) which report that among 101 undergraduates the large majority self-reported to read and learn more when they use printed format despite the fact that they prefer electronically supplied readings instead of those supplied in printed form. On the other side, Rockinson-Szapkiw, et al (2013) conducted a study were they demonstrate that e-textbook is as effective for learning as the traditional textbook and that students who choose e-textbook had significantly higher perceived learning than students who chose to use print textbooks.

A Fractional Calculus Perspective in the Evolutionary Design of Combinational Circuits

This paper analyses the performance of a genetic algorithm (GA) in the synthesis of digital circuits using two novel approaches. The first concept consists in improving the static fitness function by including a discontinuity evaluation. The measure of variability in the error of the Boolean table has similarities with the function continuity issue in classical calculus. The second concept extends the static fitness by introducing a fractional-order dynamical evaluation.