A colorimetric competitive displacement assay for the evaluation of catalytic peptides

An assay based on competitive dye displacement was adapted to detect the formation of aldol product in crude reaction mixtures, and was used to evaluate minimal peptide aldol catalysts.</p

Remainers are nostalgic too: An exploration of attitudes towards the past and Brexit preferences

Nostalgia had a prominent place in the Brexit Referendum campaign, epitomized by Nigel Farage carrying around with him an old-fashioned blue British passport on the campaign trail. In this paper, we seek to examine British attitudes towards the past through a new survey instrument administered online in July and August 2018 (N = 3,000). We empirically establish two dimensions of nostalgia that are differentially associated with political preferences. We conclude that it is the substance of the nostalgia that matters, not the looking towards the past per se

The basal epithelial marker P-cadherin associates with breast cancer cell populations harboring a glycolytic and acid-resistant phenotype

"BMC Cancer 2014 14:734"BACKGROUND: Cancer stem cells are hypoxia-resistant and present a preponderant glycolytic metabolism. These characteristics are also found in basal-like breast carcinomas (BLBC), which show increased expression of cancer stem cell markers.Recently, we demonstrated that P-cadherin, a biomarker of BLBC and a poor prognostic factor in this disease, mediates stem-like properties and resistance to radiation therapy. Thus, the aim of the present study was to evaluate if P-cadherin expression was associated to breast cancer cell populations with an adapted phenotype to hypoxia. METHODS: Immunohistochemistry was performed to address the expression of P-cadherin, hypoxic, glycolytic and acid-resistance biomarkers in primary human breast carcinomas. In vitro studies were performed using basal-like breast cancer cell lines. qRT-PCR, FACS analysis, western blotting and confocal microscopy were used to assess the expression of P-cadherin after HIF-1a stabilization, achieved by CoCl2 treatment. siRNA-mediated knockdown was used to silence the expression of several targets and qRT-PCR was employed to evaluate the effects of P-cadherin on HIF-1a signaling. P-cadherin high and low breast cancer cell populations were sorted by FACS and levels of GLUT1 and CAIX were assessed by FACS and western blotting. Mammosphere forming efficiency was used to determine the stem cell activity after specific siRNA-mediated knockdown, further confirmed by western blotting. RESULTS: We demonstrated that P-cadherin overexpression was significantly associated with the expression of HIF-1a, GLUT1, CAIX, MCT1 and CD147 in human breast carcinomas. In vitro, we showed that HIF-1a stabilization was accompanied by increased membrane expression of P-cadherin and that P-cadherin silencing led to a decrease of the mRNA levels of GLUT1 and CAIX. We also found that the cell fractions harboring high levels of P-cadherin were the same exhibiting more GLUT1 and CAIX expression. Finally, we showed that P-cadherin silencing significantly decreases the mammosphere forming efficiency in the same range as the silencing of HIF-1a, CAIX or GLUT1, validating that all these markers are being expressed by the same breast cancer stem cell population. CONCLUSIONS: Our results establish a link between aberrant P-cadherin expression and hypoxic, glycolytic and acid-resistant breast cancer cells, suggesting a possible role for this marker in cancer cell metabolismo.This work was funded by FEDER funds through the COMPETE Program (Programa Operacional Factores de Competitividade) and by national funds through FCT (Portuguese Foundation for Science and Technology, Portugal), mainly in the context of the scientific project PTDC/SAU-GMG/120049/2010-FCOMP-01-0124-FEDER-021209, and partially by PTDC/SAU-FCF/104347/2008. FCT funded the research grants of BS (SFRH/BD/69353/2010), ASR (SFRH/BPD/75705/2011), ARN (grant from the project PTDC/SAU-GMG/120049/2010), CP (SFRH/BPD/69479/2010), AV (SFRH/BPD/90303/2012), as well as JP, with Programa Ciencia 2007 (Contratacao de Doutorados para o SCTN - financiamento pelo POPH - QREN - Tipologia 4.2 - Promocao do Emprego Cientifico, comparticipado pelo Fundo Social Europeu e por fundos nacionais do MCTES) and Programa IFCT (FCT Investigator). IPATIMUP is an Associate Laboratory of the Portuguese Ministry of Science, Technology and Higher Education and is partially supported by FCT

The relationship between parent and child dysfunctional beliefs about sleep and child sleep

Cognitive theories emphasise the role of dysfunctional beliefs about sleep in the development and maintenance of sleep-related problems (SRPs). The present research examines how parents' dysfunctional beliefs about children's sleep and child dysfunctional beliefs about sleep are related to each other and to children's subjective and objective sleep. Participants were 45 children aged 11 -12 years and their parents. Self-report measures of dysfunctional beliefs about sleep and child sleep were completed by children, mothers and fathers. Objective measures of child sleep were taken using actigraphy. The results showed that child dysfunctional beliefs about sleep were correlated with father (r=.43, p<.05) and mother (r=.43, p<.05) reported child SRPs, and with Sleep Onset Latency (r=.34, p<.05). Maternal dysfunctional beliefs about child sleep were related to child SRPs as reported by mothers (r=.44, p<.05), and to child dysfunctional beliefs about sleep (r=.37, p<.05). Some initial evidence was found for a mediation pathway in which child dyfunctional beliefs mediate the relationship between parent dysfunctional beliefs and child sleep. The results support the cognitive model of SRPs and contribute to the literature by providing the first evidence of familial aggregation of dysfunctional beliefs about sleep

Reactive and Regulative Temperament in Youths: Psychometric Evaluation of the Early Adolescent Temperament Questionnaire-Revised

The present study examined the psychometric properties of the self-report version of the Early Adolescent Temperament Questionnaire-Revised (EATQ-R), which is a scale for measuring reactive and regulative temperament traits, in a large sample of children and adolescents (N = 1,055). The results indicated that the internal consistency was acceptable for most EATQ-R temperament scales. Further, principal components analysis of the instrument yielded a structure with nine components, which generally reflected the temperament scales of the EATQ-R. The test–retest stability of the scale was moderate to good, whereas the parent–child agreement was rather low. Finally, the scale correlated in a theoretically meaningful way with children’s self-reports of personality and psychopathology. It can be concluded that the EATQ-R is a useful scale for measuring aspects of reactive and regulative temperament in children and adolescents, although there is certainly room for improving the instrument

Glycolytic and Non-glycolytic Functions of Mycobacterium tuberculosis Fructose-1,6-bisphosphate Aldolase, an Essential Enzyme Produced by Replicating and Non-replicating Bacilli

The search for antituberculosis drugs active against persistent bacilli has led to our interest in metallodependent class II fructose- 1,6-bisphosphate aldolase (FBA-tb), a key enzyme of gluconeogenesis absent from mammalian cells. Knock-out experiments at the fba-tb locus indicated that this gene is required for the growth of Mycobacterium tuberculosis on gluconeogenetic substrates and in glucose-containing medium. Surface labeling and enzymatic activity measurements revealed that this enzyme was exported to the cell surface of M. tuberculosis and produced under various axenic growth conditions including oxygen depletion and hence by non-replicating bacilli. Importantly, FBA-tb was also produced in vivo in the lungs of infected guinea pigs and mice. FBA-tb bound human plasmin(ogen) and protected FBA-tb-bound plasmin from regulation by α 2-antiplasmin, suggestive of an involvement of this enzyme in host/pathogen interactions. The crystal structures of FBA-tb in the native form and in complex with a hydroxamate substrate analog were determined to 2.35- and 1.9-Å resolution, respectively. Whereas inhibitor attachment had no effect on the plasminogen binding activity of FBA-tb, it competed with the natural substrate of the enzyme, fructose 1,6-bisphosphate, and substantiated a previously unknown reaction mechanism associated with metallodependent aldolases involving recruitment of the catalytic zinc ion by the substrate upon active site binding. Altogether, our results highlight the potential of FBA-tb as a novel therapeutic target against both replicating and non-replicating bacilli.Fil: Santangelo, María de la Paz. State University of Colorado - Fort Collins; Estados Unidos. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gest, Petra M.. State University of Colorado - Fort Collins; Estados UnidosFil: Guerin, Marcelo E.. Universidad del País Vasco; EspañaFil: Coinçon, Mathieu. University of Montreal; CanadáFil: Pham, Ha. State University of Colorado - Fort Collins; Estados UnidosFil: Ryan, Gavin. State University of Colorado - Fort Collins; Estados UnidosFil: Puckett, Susan E.. Cornell University; Estados UnidosFil: Spencer, John S.. State University of Colorado - Fort Collins; Estados UnidosFil: Gonzalez Juarrero, Mercedes. State University of Colorado - Fort Collins; Estados UnidosFil: Daher, Racha. Universite de Paris XI. Institut de Chimie Moléculaire et des Matériaux d'Orsay; FranciaFil: Lenaerts, Anne J.. State University of Colorado - Fort Collins; Estados UnidosFil: Schnappinger, Dirk. Cornell University; Estados UnidosFil: Therisod, Michel. Universite de Paris XI. Institut de Chimie Moléculaire et des Matériaux d'Orsay; FranciaFil: Ehrt, Sabine. Cornell University; Estados UnidosFil: Sygusch, Jurgen. University of Montreal; CanadáFil: Jackson, Mary. State University of Colorado - Fort Collins; Estados Unido

‘Birds of a feather’? Assessing the prevalence of anti-immigration attitudes among the far right electorate

This article focuses on the prevalence of anti-immigration attitudes among the far-right electorate. Drawing on the distinction between the predictive power of immigration concerns, and the question of how widespread these concerns are among the far-right voter pool, we proceed in two steps. First, we assess the extent to which anti-immigration attitudes are a necessary condition for voting far right; and second we examine whether far right voters with different levels of anti-immigration attitudes exhibit similar individual and attitudinal characteristics. Using data from the 8th wave of the European Social Survey (ESS) we find that, surprisingly, anti-immigration attitudes are not a necessary condition for voting for the far right as approximately one third of far-right voters have no concerns over immigration. We further show that far-right voters with different levels of immigration concerns have different profiles when it comes to other predictors of the far right-vote including ideological affinity, attachment to the EU and government satisfaction. Our contribution is significant as we suggest that there are different routes to voting for the far right by groups with different grievances, including non- immigration related

Model International Mobility Convention

While people are as mobile as they ever were in our globalized world, the movement of people across borders lacks global regulation. This leaves many refugees in protracted displacement and many migrants unprotected in irregular and dire situations. Meanwhile, some states have become concerned that their borders have become irrelevant. International mobility—the movement of individuals across borders for any length of time as visitors, students, tourists, labor migrants, entrepreneurs, long-term residents, asylum seekers, or refugees—has no common definition or legal framework. To address this key gap in international law, and the growing gaps in protection and responsibility that are leaving people vulnerable, the "Model International Mobility Convention" proposes a framework for mobility with the goals of reaffirming the existing rights afforded to mobile people (and the corresponding rights and responsibilities of states) as well as expanding those basic rights where warranted. In 213 articles divided over eight chapters, the Convention establishes both the minimum rights afforded to all people who cross state borders as visitors, and the special rights afforded to tourists, students, migrant workers, investors and residents, forced migrants, refugees, migrant victims of trafficking and migrants caught in countries in crisis. Some of these categories are covered by existing international legal regimes. However, in this Convention these groups are for the first time brought together under a single framework. An essential feature of the Convention is that it is cumulative. This means, for the most part, that the chapters build on and add rights to the set of rights afforded to categories of migrants covered by earlier chapters. The Convention contains not only provisions that afford rights to migrants and, to a lesser extent, States (such as the right to decide who can enter and remain in their territory). It also articulates the responsibilities of migrants vis-à-vis States and the rights and responsibilities of different institutions that do not directly respond to a right held by migrants