Liquid crystal anchoring transitions on aligning substrates processed by plasma beam

We observe a sequence of the anchoring transitions in nematic liquid crystals (NLC) sandwiched between the hydrophobic polyimide substrates treated with the plasma beam. There is a pronounced continuous transition from homeotropic to low tilted (nearly planar) alignment with the easy axis parallel to the incidence plane of the plasma beam (the zenithal transition) that takes place as the exposure dose increases. In NLC with positive dielectric anisotropy, a further increase in the exposure dose results in in-plane reorientation of the easy axis by 90 degrees (the azimuthal transition). This transition occurs through the two-fold degenerated alignment characteristic for the second order anchoring transitions. In contrast to critical behavior of anchoring, the contact angle of NLC and water on the treated substrates monotonically declines with the exposure dose. It follows that the surface concentration of hydrophobic chains decreases continuously. The anchoring transitions under consideration are qualitatively interpreted by using a simple phenomenological model of competing easy axes which is studied by analyzing anchoring diagrams of the generalized polar and non-polar anchoring models.Comment: revtex4, 18 pages, 10 figure

Towards a competitive and sustainable OA market in Europe - A study of the open access market and policy environment

Deliverable 5.3 of OpenAIRE WP5: FP7 Post Grant Gold Open Access Pilot. This deliverable consists of a study and an annex - and it will be supplemented by a roadmap in May 2017. This study considers the economic factors contributing to the current state of the open-access publishing market, and evaluates the potential for European policymakers to enhance market competition and sustainability in parallel to increasing access. It was commissioned within the scope of the OpenAIRE FP7 Post-Grant Open Access Pilot, and it will be accompanied by a Roadmap document developed with inputs from an expert workshop to be held in The Hague on 20 April 2017. In accordance with the project brief, the study aims to: Explore the current status of the OA publishing market Analyse existing OA publishing business models Evaluate how different national and international policies are complementing each other as a means to achieve a transition to OA Evaluate the impact of the Framework Programme 7 Post-grant OA pilot and its implications for future similar initiatives and the transition to OA. Provide a roadmap leading to a sustainable and competitive market The transition to open access concerns all kinds of academic research outputs, including monographs, journal articles, and data. This study focuses on open access to peer-reviewed research articles, which constitute the bulk of the market and the primary mechanism through which research is disseminated across disciplinary communities and beyond. This report is supplemented by an Annex containing a mid-term evaluation of the FP7 Post-Grant Open Access Pilot.This report will be accompanied by a Roadmap document developed with inputs from an expert workshop to be held in The Hague on 20 April 201

MITOCHONDRIAL MASS, DISTRIBUTION AND ACTIVITY DURING SEA URCHIN OOGENESIS

The sea urchin egg is a favourite model for studies of the molecular biology and physiology of fertilization and early development, yet we know sparingly little of its oocytes and of mitochondria behaviour during oogenesis. The process of oogenesis in most echinoderms is asynchronous so each ovary lobe has hundreds of oocytes at all stages of development. At the beginning of oogenesis, the oocyte is about 10 \ub5m in diameter. During the vitellogenic phase of oogenesis, the oocyte accumulate yolk proteins and grow to ten times their original size to 80 to 100 \ub5m in sea urchins. The oocyte, arrested at the prophase of the first meiotic division, is apparent with its large nucleus, the germinal vesicle (GV), containing a prominent nucleolus. Echinoid (such as sea urchin) and Holothurian oocytes complete meiotic maturation prior to fertilization, distinct from other echinoderms and almost all others animals. As maturation progresses, it occurs the GV breaks down (GVBD). These eggs may then be stored for weeks to months within the female before they are spawned, and the proportion of eggs in the ovary increases from early to late season, as the numbers of oocytes decline [1]. Mitochondria, generally known as the powerhouses of eukaryotic cells, play a primary role in cellular energetic metabolism, homeostasis and death. These organelles, with their multicopy genome maternally inherited, are directly involved at several levels in the reproductive process since their functional status influences the quality of oocytes and contributes to the process of fertilization and embryonic development. It has been demonstrated that the number of maternal mitochondria is sufficient to support development until late stages without new synthesis of mitochondrial DNA or production of new organelles [2]. During embryogenesis mitochondrial mass does not change, whereas mitochondrial respiration increases [3]. The behaviour of these organelles during oogenesis remains at moment unclear. In the present paper we studied, by Confocal Laser Scanning Microscopy tecnologies (CLSM), the mass and distribution, the activity and the DNA content of sea urchin Paracentrotus lividus mitochondria during oogenesis, by in vivo incubating oocytes of different size with cell-permeant probes specific for mitochondria and for DNA and by immunodetection of hsp60 chaperonine, a well known mitochondrial marker. In particular the oocytes were grouped in six classes: < 10, 20/30, 40/50, 60/70, 80/90 \ub5m, and 90 \ub5m ovulated egg, on the base of diameters. Microscopic observations were performed capturing 2 \ub5m thick layers of oocytes. Of the several thousands oocytes we observed, 20 for each different oogenesis stage were analyzed and processed. In order to interpret results and to draw unequivocal conclusions, we measured by IMAGE J software analysis the intensity values of fluorescent signals, as suggested in Agnello et al 2008 [4]. The mitochondria of oocytes with a diameter between 20 and 70 \ub5m, appeared to give rise to clusters that disappear in that of 80 \ub5m. In the oocytes between 60 and 90 \ub5m the red fluorescence seems to be more evident around the germinal vesicle (the merge tends to red), suggesting an increasing oxidative phosphorylation activity. In the ovulated eggs, red and green fluorescence are uniformly distributed suggesting that mitochondria are dispersed in the cytoplasm. In addition the merge of green and red colours shows that the whole mitochondrial population is consuming oxygen at the same level (the resulting colours tends to yellow), figure 1. In order to calculate the total mitochondrial mass and activity we integrated the values of pixel intensities for all captured sections and used the arithmetic means to draw a statistical analysis. Results suggest a parallel rise of mitochondrial mass and activity, suggesting that the amount and activity of organelles change remarkably during oogenesis. Figure 1. shows the distribution of hsp60 protein, detected by immunofluorescence analysis (A), the mitochondrial and genomic DNA, after in vivo incubation with PicoGreen probe (B) and the merge of green and red fluorescence signal, respectively due to mitochondrial mass and activity, after in vivo incubation with Mitotraker Green and Orange (C). The size of the oocytes reported is 80 \ub5m. Results suggest that mitochondria are actively duplicating and that mitochondrial DNA is replicating during the different oogenesis phases. It is noteworthy that around the germinal vesicle, especially in the larger oocytes, next to the germinal vesicle breakdown, the organelles are more active in oxygen consumption, probably due to the major energetic needing in this key moment of gametogenesis. [1] Wessel G.M., Voronina E., and Brooks J.M. (2004) Obtaining and handling echinoderm oocytes. In \u201cMethods in Cell Biology\u201d, Elsevier. Vol.74, Chapter 5, pp. 87-114. [2] Matsumoto L., Kasamatsu H., Pik\ub4o L. and Vinograd J. (1974) Mitochondrial DNA replication in sea urchin oocytes. J. Cell Biol. 63: 146\u2013159. [3] Morici G., Agnello M., Spagnolo F., Roccheri M.C., Di Liegro C.M. and Rinaldi A.M. (2007) Confocal microscopy study of the distribution, content and activity of mitochondria during Paracentrotus lividus development. Journal of Microscopy. 228: 165-173. [4] Agnello M., Morici G., Rinaldi A.M. (2008) A method for measuring mitochondrial mass and activity . Cytotechnology. 56: 145-149. Maria Carmela Roccheri: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche. Universit\ue0 degli Studi di Palermo, Viale delle Scienze Ed.16, Palermo, Italy; tel: 09123897414; e-mail: [email protected]

Functionalization of Organic Semiconductor Crystals via the Diels-Alder Reaction

A surface adlayer is generated on organic single crystals (tetracene and rubrene) using the site specific Diels–Alder reaction and a series of vapor phase dienophiles. X-ray photoelectron spectroscopy (XPS) confirms adsorption on the surfaces of tetracene and rubrene and mass spectrometry demonstrates the reaction’s applicability to a range of dienophiles

APOPTOTIC ANALYSIS OF CUMULUS CELLS FOR THE SELECTION OF COMPETENT OOCYTES TO BE FERTILIZED BY INTRACYTOPLASMIC SPERM INJECTION (ICSI)

Oocyte quality is one of the main factors for the success of in vitro fertilization protocols. Apoptosis is known to affect oocyte quality and may impair subsequent embryonic development and implantation. The aim of this study was to investigate the apoptosis rate of single and pooled cumulus cells of cumulus cell\u2013oocyte complexes (COCs), as markers of oocyte quality, prior to intracytoplasmatic sperm injection (ICSI).We investigated the apoptosis rate by TUNEL assay (DNA fragmentation) and caspase-3 immunoassay of single and pooled cumulus cells of COCs. The results showed that DNA fragmentation in cumulus cells was remarkably lower in patients who achieved a pregnancy than in those who did not. Cumulus cell apoptosis rate could be a marker for the selection of the best oocytes to be fertilized by intracytoplasmatic sperm injection

Autophagy as a defense strategy against stress: focus on Paracentrotus lividus sea urchin embryos exposed to cadmium

Autophagy is used by organisms as a defense strategy to face environmental stress. This mechanism has been described as one of the most important intracellular pathways responsible for the degradation and recycling of proteins and organelles. It can act as a cell survival mechanism if the cellular damage is not too extensive or as a cell death mechanism if the damage/stress is irreversible; in the latter case, it can operate as an independent pathway or together with the apoptotic one. In this review, we discuss the autophagic process activated in several aquatic organisms exposed to different types of environmental stressors, focusing on the sea urchin embryo, a suitable system recently included into the guidelines for the use and interpretation of assays to monitor autophagy. After cadmium (Cd) exposure, a heavy metal recognized as an environmental toxicant, the sea urchin embryo is able to adopt different defense mechanisms, in a hierarchical way. Among these, autophagy is one of the main responses activated to preserve the developmental program. Finally, we discuss the interplay between autophagy and apoptosis in the sea urchin embryo, a temporal and functional choice that depends on the intensity of stress conditions

APOPTOSIS RATE IN CUMULUS CELLS AS POSSIBLE MOLECULAR BIOMARKER FOR OOCYTE COMPETENCE.

Several lines of evidence showed that apoptosis rate of cumulus cells in oocytes derived by assisted reproductive technologies could be used as an indicator of fertilizing gamete quality. Aim of the study was to investigate the effects of three different ovarian stimulation protocols on the biological and clinical outcome in hyporesponder patients. Collected data showed a higher significant rate of DNA fragmentation index (DFI) in U group (patients treated with Highly Purified human Menopausal Gonadotrophin) than in P group (treated with recombinant human Follicle Stimulating Hormone (r-hFSH) combined with recombinant human Luteinizing Hormone (r-hLH)). Both groups R (treated with r-hFSH alone) and P showed a significant increase in collected and fertilized oocytes number, embryo quality number. This study showed that combined r-hFSH/r-hLH therapy could represent the best pharmacological strategy for controlled ovarian stimulation and suggests to use DFI as a biomarker of ovarian function in hyporesponder patients

Toxicity induced by Gadolinium ions on sea urchin embryos: comparison among phylogenetically distant species and focus on stress response and skeletogenesis.

Pharmaceuticals are a class of emerging environmental contaminants. Gadolinium (Gd) is a lanthanide metal whose chelates are employed as contrast agents for magnetic resonance imaging, and subsequently released into the aquatic environment. We investigated the effects of exposure to sublethal Gd concentrations on the development of four phylogenetically and geographically distant sea urchin species: two Mediterranean, Paracentrotus lividus and Arbacia lixula, and two from Australia, Heliocidaris tuberculata and Centrostephanus rodgersii. Sensitivity to Gd greatly varied, with EC50 ranging from 56 nM to 132 µM across the four species. Measures of the Gd and Ca content inside embryos showed a time- and dose-dependent increase in Gd, in parallel with a reduction in Ca. In all the four species, we observed a general delay of embryo development at 24h post-fertilization, and a strong inhibition of skeleton growth at 48h. Further experiments were carried out on P. lividus embryos: RT-PCR gene expression analysis showed the misregulation of several genes implicated both in the skeletogenic and the left-right axis specification networks. WB analysis showed an increase of the LC3 autophagic marker at 24 and 48h. Confocal microscopy studies confirmed the increased number of autophagosomes and autophagolysosomes and showed no apoptotic induction. The results show the hazard of Gd in the marine environment, indicating that Gd is able to affect three different levels in sea urchin embryos: morphogenesis, stress response such as autophagy, and gene expression. Results highlight that pollution assays based on only one species can be misleading with respect to hazard risk assessment

Resisted adduction in hip neutral is a superior provocation test to assess adductor longus pain:an experimental pain study

The criterion of long-standing groin pain diagnoses in athletes usually relies on palpation and clinical tests. An experimental pain model was developed to examine the clinical tests under standardized conditions. Pain was induced by hypertonic saline injected into the proximal adductor longus (AL) tendon or rectus femoris (RF) tendon in 15 healthy male participants. Isotonic saline was injected contralaterally as a control. Pain intensity was assessed on a visual analog scale (VAS). Resisted hip adduction at three different angles and trunk flexion were completed before, during, and after injections. Pain provocation in the presence of experimental pain was recorded as a true positive compared with pain provocation in the non-pain conditions. Similar peak VAS scores were found after hypertonic saline injections into the AL and RF and both induced higher VAS scores than isotonic saline (P<0.01). Adduction at 0° had the greatest positive likelihood ratio (+LR=2.8, 95%CI: 1.09-7.32) with 45° (-LR=0.0, 95%CI: 0.00-1.90) and 90° (-LR=0.0, 95%CI: 0.00-0.94) having the lowest negative LR. This study indicates that the 0° hip adduction test resisted at the ankles optimizes the diagnostic procedure without compromising diagnostic capacity to identify experimental groin pain. Validation in clinical populations is warranted