Protein-responsive ribozyme switches in eukaryotic cells

Genetic devices that directly detect and respond to intracellular concentrations of proteins are important synthetic biology tools, supporting the design of biological systems that target, respond to or alter specific cellular states. Here, we develop ribozyme-based devices that respond to protein ligands in two eukaryotic hosts, yeast and mammalian cells, to regulate the expression of a gene of interest. Our devices allow for both gene-ON and gene-OFF response upon sensing the protein ligand. As part of our design process, we describe an in vitro characterization pipeline for prescreening device designs to identify promising candidates for in vivo testing. The in vivo gene-regulatory activities in the two types of eukaryotic cells correlate with in vitro cleavage activities determined at different physiologically relevant magnesium concentrations. Finally, localization studies with the ligand demonstrate that ribozyme switches respond to ligands present in the nucleus and/or cytoplasm, providing new insight into their mechanism of action. By extending the sensing capabilities of this important class of gene-regulatory device, our work supports the implementation of ribozyme-based devices in applications requiring the detection of protein biomarkers

Everyday pain, analgesic beliefs and analgesicbehaviours in Europe and Russia: An epidemiologicalsurvey and analysis

Беше проведено анкетно проучване в осем държави, в т.ч. Белгия, Германия, Великобритания, Италия, Полша, Русия и Испания. Оценена беше честотата на оплакване от болка при 8 506 души (52% жени), както и използването на аналгетици и нагласите спрямо тях. Предварителните анализи потвърждават високата честота на оплаквания от болка, като 70% от анкетираните посочват, че поне веднъж в месеца изпитват такава. Най-често се съобщава за главоболие и болки в гърба, a болшинството анкетирани лица - 77%, споделят, че използват аналгетични средства, когато страдат от болка. Предмет на по-нататъшните анализи бяха различията по пол, статус на заетост, държава по местоживеене и възраст. В сравнение с мъжете, жените по-често изпитват болка, употребата на аналгетици сред тях е по-голяма, както и притесненията им във връзка с тази употреба, като проявяват склонност да базират употребата върху своите познания относно лекарствените продукти. Безработните също съобщават за по-голямо използване на аналгетици и повече притеснения във връзка с това, сравнено със заетите лица. Като цяло при хората, живеещи в Русия и Полша, се отчита по-рядко болка и по-малко използване на обезболяващи средства в сравнение с останалата част на Европа. Те също така се тревожат повече във връзка с аналгетиците и по отношение на това, че използването на аналгетици от тяхна страна е слабо свързано с познания относно лекарствата. Отчитането на фактора „възраст` показва, че при по-младите епизодите на болка са по-малко, употребата на аналгетици - по-честа, a притесненията относно аналгетиците са по-слаби в сравнение с по-възрастните. Също така, по-младите са по-склонни да използват аналгетици въз основа на своите познания за продукта. Тези резултати повтарят съществуващи констатации във връзка с честотата на болката. Дават и нови сведения за различията при изпитването на болка, използването на аналгетици и нагласите към аналгетиците в Европа и Русия.

Interaction between src family kinases and rho-kinase in agonist-induced Ca2+-sensitization of rat pulmonary artery

Aims: We investigated the role of src family kinases (srcFK) in agonist-mediated Ca2+-sensitization in pulmonary artery and whether this involves interaction with the rho/rho-kinase pathway. Methods and results: Intra-pulmonary arteries (IPAs) and cultured pulmonary artery smooth muscle cells (PASMC) were obtained from rat. Expression of srcFK was determined at the mRNA and protein levels. Ca2+-sensitization was induced by prostaglandin F(2 alpha) (PGF(2 alpha)) in alpha-toxin-permeabilized IPAs. Phosphorylation of the regulatory subunit of myosin phosphatase (MYPT-1) and of myosin light-chain-20 (MLC20) and translocation of rho-kinase in response to PGF(2 alpha) were also determined. Nine srcFK were expressed at the mRNA level, including src, fyn, and yes, and PGF(2 alpha) enhanced phosphorylation of three srcFK proteins at tyr-416. In alpha-toxin-permeabilized IPAs, PGF(2 alpha) enhanced the Ca2+-induced contraction (pCa 6.9) approximately three-fold. This enhancement was inhibited by the srcFK blockers SU6656 and PP2 and by the rho-kinase inhibitor Y27632. Y27632, but not SU6656 or PP2, also inhibited the underlying pCa 6.9 contraction. PGF(2 alpha) enhanced phosphorylation of MYPT-1 at thr-697 and thr-855 and of MLC20 at ser-19. This enhancement, but not the underlying basal phosphorylation, was inhibited by SU6656. Y27632 suppressed both basal and PGF(2 alpha)-mediated phosphorylation. The effects of SU6656 and Y27632, on both contraction and MYPT-1 and MLC20 phosphorylation, were not additive. PGF(2 alpha) triggered translocation of rho-kinase in PASMC, and this was inhibited by SU6656. Conclusions: srcFK are activated by PGF(2 alpha) in the rat pulmonary artery and may contribute to Ca2+-sensitization and contraction via rho-kinase translocation and phosphorylation of MYPT-1

Wear of human teeth: a tribological perspective

The four main types of wear in teeth are attrition (enamel-on-enamel contact), abrasion (wear due to abrasive particles in food or toothpaste), abfraction (cracking in enamel and subsequent material loss), and erosion (chemical decomposition of the tooth). They occur as a result of a number of mechanisms including thegosis (sliding of teeth into their lateral position), bruxism (tooth grinding), mastication (chewing), toothbrushing, tooth flexure, and chemical effects. In this paper the current understanding of wear of enamel and dentine in teeth is reviewed in terms of these mechanisms and the major influencing factors are examined. In vitro tooth wear simulation and in vivo wear measurement and ranking are also discussed

The Forgotten Side of Partisanship: Negative Party Identification in Four Anglo-American Democracies

Early studies of electoral behavior proposed that party identification could be negative as well as positive. Over time, though, the concept became mostly understood as a positive construct. The few studies that took negative identification into account tended to portray it as a marginal factor that went “hand-in-hand” with positive preferences. Recent scholarship in psychology reaffirms, however, that negative evaluations are not simply the bipolar opposite of positive ones. This article considers negative party identification from this standpoint, and evaluates its impact in recent national elections in Australia, Canada, New Zealand, and the United States. Our findings highlight the autonomous power of negative partisanship. They indicate as well that ideology has an influence on both positive and negative partisan identification

Correlational analysis and predictive validity of psychological constructs related with pain in fibromyalgia

<p>Abstract</p> <p>Background</p> <p>Fibromyalgia (FM) is a prevalent and disabling disorder characterized by a history of widespread pain for at least three months. Pain is considered a complex experience in which affective and cognitive aspects are crucial for prognosis. The aim of this study is to assess the importance of pain-related psychological constructs on function and pain in patients with FM.</p> <p>Methods</p> <p>Design</p> <p>Multicentric, naturalistic, one-year follow-up study.</p> <p><it>Setting and study sample</it>. Patients will be recruited from primary care health centres in the region of Aragon, Spain. Patients considered for inclusion are those aged 18-65 years, able to understand Spanish, who fulfil criteria for primary FM according to the American College of Rheumatology, with no previous psychological treatment.</p> <p>Measurements</p> <p>The variables measured will be the following: main variables (pain assessed with a visual analogue scale and with sphygmomanometer and general function assessed with Fibromyalgia Impact Questionnaire, and), psychological constructs (pain catastrophizing, pain acceptance, mental defeat, psychological inflexibility, perceived injustice, mindfulness, and positive and negative affect), and secondary variables (sociodemographic variables, anxiety and depression assessed with Hospital Anxiety and Depression Scale, and psychiatric interview assessed with MINI). Assessments will be carried at baseline and at one-year follow-up.</p> <p>Main outcome</p> <p>Pain Visual Analogue Scale.</p> <p>Analysis</p> <p>The existence of differences in socio-demographic, main outcome and other variables regarding pain-related psychological constructs will be analysed using Chi Square test for qualitative variables, or Student <it>t </it>test or variance analysis, respectively, for variables fulfilling the normality hypothesis. To assess the predictive value of pain-related psychological construct on main outcome variables at one-year follow-up, use will be made of a logistic regression analysis adjusted for socio-demographic and clinical variables. A Spearman Rho non-parametric correlation matrix will be developed to determine possible overlapping between pain-related psychological constructs.</p> <p>Discussion</p> <p>In recent years, the relevance of cognitive and affective aspects for the treatment of chronic pain, not only in FM but also in other chronic pain diseases, has been widely acknowledged. However, the relative importance of these psychological constructs, the relationship and possible overlapping between them, or the exact meaning of them in pain are not enough known.</p

Working with pain : sustainable work participation of workers with chronic nonspecific musculoskeletal pain

Dit proefschrift is een van de eerste studies specifiek gericht op mensen die blijven werken met chronische pijn aan het bewegingsapparaat. Unieke kennis over duurzame arbeidsparticipatie van werknemers met chronische pijn werd vergaard. Doel van dit promotieonderzoek was meer inzicht te krijgen in de groep mensen die werkt met pijn en te achterhalen hoe zij in staat zijn ondanks hun klachten te blijven werken. In het proefschrift staan kenmerken en determinanten beschreven van werknemers die doorwerken met chronische pijn, waardoor een completer beeld is ontstaan van arbeidsparticipatie bij mensen met chronische pijn aan het bewegingsapparaat. Uit de vergelijking van mensen die doorwerken ondanks chronische pijn en mensen met arbeidsverzuim die in revalidatiebehandeling komen met chronische pijn blijkt dat deze groepen op diverse factoren significant verschillen. In het onderzoek werd onder andere aangetoond dat de motivatie voor werk, zelfmanagementvaardigheden en het belang dat wordt toegekend aan pijn, belangrijke factoren zijn die werken met chronische pijn faciliteren. Chronische pijn op zichzelf is vaak niet de reden voor arbeidsverzuim, maar meestal spelen persoonlijke- en omgevingsfactoren daarin een beslissende rol. Deze factoren kunnen dienen als aangrijpingspunt voor het verhogen van duurzame inzetbaarheid en preventie van arbeidsverzuim van mensen met chronische pijn aan het bewegingsapparaat. De effectieve manier waarop deelnemende werknemers in het onderzoek met hun pijn omgingen en productief bleven, kan anderen inspireren aan het werk te blijven. Daarnaast biedt het onderzoek een nieuw referentiekader voor de bedrijfs-, verzekerings-, en revalidatiegeneeskunde. This thesis was one of the first studies that focused specifically on people who continued work with chronic nonspecific musculoskeletal pain (CMP), and collected (identified) unique data concerning sustainable work participation of workers with CMP. It provides a large range of characteristics of workers with CMP who continued work despite pain, which has added to our understanding of sustainable work participation in people suffering from CMP. Comparison of workers who continued work with CMP with sick listed workers with CMP admitted for rehabilitation revealed that these groups differ significantly on several factors. In this thesis, evidence was found that the workers’ motivation to work, self-management skills, and the attributed importance of pain on their (working) lives are important factors to manage staying at work with CMP. It is recommended to be aware of the fact that CMP standing on itself is often not the reason for sick leave and disability, but regularly personal and environmental factors play an additional decisive role. Because these factors can be influenced, they offer opportunity to promote staying at work. In the process of guiding workers back to work, the results of the project ‘Working with pain’ may be used. The findings of this thesis potentially contribute to promotion of sustained work participation and prevention of sick-leave in workers with CMP. The effective way workers in this project coped with CMP and remained productive, may inspire others in their efforts to stay work. Finally, this thesis offers a new reference for rehabilitation-, occupational-,and insurance medicine.

Benchmark models for the analysis and control of small-signal oscillatory dynamics in power systems

This paper summarizes a set of six benchmark systems for the analysis and control of electromechanical oscillations in power systems, recommended by the IEEE Task Force on Benchmark Systems for Stability Controls of the Power System Dynamic Performance Committee. The benchmark systems were chosen for their tutorial value and particular characteristics leading to control the system design problems relevant to the research community. For each benchmark, the modeling guidelines are provided, along with eigenvalues and time-domain results produced with at least two simulation softwares, and one possible control approach is provided for each system as well. Researchers and practicing engineers are encouraged to use these benchmark systems when assessing new oscillation damping control strategies