Interaction of he Epsin N-Terminal Homology domain (ENTH) with artificial membranes as a function of lateral tension

Proteine formen während der Endozytose planare Membranen zu gekrümmten Vesikeln um. Im ersten Schritt dieses Prozesses bindet das Protein Epsin an das Rezeptorlipid Phosphatidylinositol-4,5-bisphosphat (PIP2) und ein vorher ungeordneter Bereich am N-Terminus von Epsin, die epsin N-terminal homology domain (ENTH), bildet eine α-Helix, welche in die Membran insertiert. Im Rahmen dieser Arbeit wurde die Wechselwirkung von ENTH mit PIP2-haltigen Lipiddoppelschichten unter Verwendung von bottom up Modellsystemen charakterisiert. Die Affinität von ENTH zu PIP2 wurde für verschiedene Lipidzusammensetzungen und Membrangeometrien untersucht, wobei unabhängig von Lipidzusammensetzung und Membrantopologie ähnliche Bindungskonstanten im hohen nanomolaren Bereich bestimmt wurden. Ausgestülpte porenüberspannende Membranen wurden als Modellsystem etabliert, um die Fähigkeit von ENTH zur Membrankrümmung als Funktion der Lipidzusammensetzung zu charakterisieren. Die Höhe der ausgestülpten Membranen ist durch die laterale Spannung begrenzt. Verursacht durch Insertion der ENTH Helix wuchsen Membranen mit einem hohen Flächenkompressionsmodul. Im Gegensatz dazu rissen Membranen mit einem niedrigen Flächenkompressionsmodul durch die ENTH induzierte Bildung von Membrandefekten. Entgegen der Eigenschaft von ENTH Membranen zu krümmen, wurde an hochgespannten porenüberspannenden Membranen keine Membrantubulierung und -vesikulierung beobachtet. Daher wurde untersucht, ob diese Fähigkeit durch eine hohe laterale Spannung unterdrückt wird. Zu diesem Zweck wurden Riesenvesikel auf einem Glassubstrat adhäriert, wobei die Adhäsionsstärke und in Folge die laterale Spannung als Funktion der Mg2+ Konzentration eingestellt werden konnte. ENTH-induzierte Membrantubulierung konnte für Vesikel mit niedriger Spannung nachgewiesen werden und war bei höherer Spannung unterdrückt. Unabhängig von der Membranspannung wurde ein Abflachen der Vesikel nach ENTH-Zugabe beobachtet. Die Ursache hierfür wurde in der durch die insertierte Helix hervorgerufene Reduktion des Flächenkompressionsmoduls gefunden. Die insertierte Helix stört die hydrophoben Wechselwirkungen der Lipidfettsäureketten und das reduzierte Flächenkompressionsmodul verringert die zur Membrankrümmung benötigte Energie. In Kombination mit der durch die insertierte Helix erzeugten lokalen Krümmung ist dies eine molekulare Erklärung für die ENTH-initiierte Bildung eines Vesikels während der Endozytose.During endocytosis, membrane remodeling of a planar membrane into a highly curved vesicle is controlled by a complex protein machinery. However, the regulatory role of physical properties such as membrane tension is highly debated. At the beginning of the formation of a vesicle during clathrin mediated endocytosis, the protein epsin binds to its receptor lipid phosphatidylinositol-4,5-bisphosphate (PIP2). Upon binding, a previously unstructured part of the epsin N-terminal homology domain (ENTH) forms an α-helix, which inserts into the membrane. Experiments using artificial lipid bilayers were performed to study the interaction of ENTH and lipid bilayers as a function of lipid composition and membrane tension. The binding affinity of ENTH to PIP2 was analyzed for different lipid compositions and membrane topologies. Similar affinities of ENTH binding to PIP2 in the high nanomolar range were measured, independent of the used lipid composition and membrane topology investigated. Protruded pore-spanning membranes were established to investigate the remodeling activity of ENTH as a function of lipid composition. Binding of ENTH to membranes having a large area compressibility modulus and a high lysis tension resulted in growth of the membrane protrusions. Binding of ENTH to membranes with a lower area compressibility modulus and lysis tension resulted in the formation of membrane defects. For all lipid compositions analyzed, no vesiculation or tubulation was observed after binding of ENTH. To analyze whether the high membrane tension of the protruded pore-spanning membranes suppresses any membrane remodeling ability of ENTH, experiments with giant unilamellar vesicles adhering to a solid support were conducted. The vesicles’ adhesion strength and lateral tension was adjusted to analyze the ENTH’s remodeling ability at lateral tensions corresponding to low and high membrane tensions found in cells. The formation of membrane tubes was observed for vesicles having a low membrane tension. Increasing the membrane tension resulted in a suppression of tube formation. Independent of the membrane tension, flattening of the vesicles was monitored after binding of ENTH. The inserted ENTH helix disturbs lipid packing, which reduces the area compressibility modulus and thus the bending rigidity of the membranes. The reduced bending rigidity lowers the energy required for the generation of membrane curvature. As the inserted helix of ENTH also splays the lipid head groups, thereby inducing local curvature, the combination of both mechanisms is expected to efficiently initiate the formation of a vesicle during clathrin mediated endocytosis

Усна історія другої світової війни на Кіровоградщині (за матеріалами етнографічних соціально-педагогічних практик студентів факультету історії та права КДПУ імені В. Винниченка 2008 - 2013 рр.)

(UA) У статті узагальнено досвід збирання та опрацювання джерел усної історії Другої світової війни на Кіровоградщині польовими етнографічними експедиціями, організованими факультетом історії та права КДПУ імені В. Винниченка.(EN) This article generalizes experience of collection and processing of oral historical sources of World War II in the Region of Kirovohrad performed by field ethnographical expeditions arranged by the Department of History and Law of the Kirovohrad State Pedagogical University named after V. Vynnytchenko

Recent Developments in European Capital Markets: Key findings from the 2018 ECMI Statistical Package. CEPS ECMI Research Report No. 14 | December 2018

This paper provides an overview of the key findings observed in the 2018 ECMI Statistical Package, a comprehensive and annually updated database on the dynamics of European and global capital markets (covering the US, Japan, China and other relevant markets). The key trends obtained from the Package on equity markets, debt securities, exchange-traded derivatives, over-the-counter derivatives and asset management are outlined in this repor

Draaman materiaalipaketti varhaiskasvattajille

Opinnäytetyömme tarkoituksena oli kehittää ja luoda draaman materiaalipaketti Kirkkonummelaisen Norlandia Päiväkodin henkilökunnalle. Materiaalipaketin avulla pyritään lisäämään draamakasvatusta päiväkodin arjessa. Opinnäytetyön idea tuli päiväkodin johtajalta, jolloin tarve materiaalipaketista tuli selkeäksi. Opinnäyteyömme yhteistyökumppanina toimivat Kirkkonummella sijaitseva Norlandia Päiväkoti ja sen henkilökunta sekä lapset. Opinnäytetyömme tavoitteena oli suunnitella Norlandia Päiväkotiin Tenava-Taaperoon materiaalipaketti draamakasvatuksesta, joka sisältää käsinukke-, varjo-, ja pöytäteatteriin liittyviä tarinoita, niihin liittyvät tarvikkeet, keskusteluaiheita sekä tarinoihin liittyvät tavoitteet. Opinnäytetyömme tavoitteena oli helpottaa ja lisätä varhaiskasvattajien draamakasvatusta, sen suunnittelua ja toteutusta. Tavoitteena on myös, että varhaiskasvattajilla on tarvittavat välineet draamakasvatukseen helposti lähettyvillä. Opinnäytetyömme tavoitteena oli, että draamakasvatus lisääntyisi päiväkodin arjessa ja toiminnoissa, sekä lapset innostuisivat myös itse käyttämään teatterinukkeja. Tavoitteena oli myös, että materiaalipaketin avulla päiväkodin työntekijät pystyisivät helpommin esittämään ja luomaan erilaisia teatteriesityksiä lasten kanssa, ja että materiaalipakettia voisi hyödyntää kaikenikäisten päiväkotilasten kanssa. Opinnäytetyön toiminnallinen osuus koostui materiaalipaketin suunnittelusta, sen toteutuksesta sekä testauksesta päiväkodin henkilökunnan sekä lasten kanssa. Materiaalipaketin toimivuutta arvioitiin päiväkodin työntekijöiden ja lasten antamien palautteiden kautta. Materiaalipaketti annetaan testauksen jälkeen päiväkodille käytettäväksi ja he voivat jatkaa materiaalipakettia vielä meidän jälkeemme. Teoreettisessa viitekehyksessä käsitellään varhaiskasvatuksen kehitystä, luovia menetelmiä, uutta varhaiskasvatussuunnitelmaa, draamakasvatusta eri varhaiskasvatussuunnitelmien näkökulmasta, sekä draamakasvatusta ja nukketeatteria yleisesti. Opinnäytetyön toiminnallinen osuus koostui materiaalipaketin suunnittelusta, sen toteutuksesta sekä testauksesta päiväkodin henkilökunnan sekä lasten kanssa. Materiaalipaketin toimivuutta arvioitiin päiväkodin työntekijöiden ja lasten antamien palautteiden kautta. Materiaalipaketti annetaan testauksen jälkeen päiväkodille käytettäväksi ja he voivat jatkaa materiaalipakettia vielä meidän jälkeemme.The purpose of this Bachelor’s thesis was to develop and create a drama-based material package for early childhood educators at a day care centre Norlandia in Kirkkonummi. Through the package, the aim is to increase drama education in the everyday functions of the care centre. The idea for this material package came from the manager of the day care centre. The material package was carried out in association with Norlandia day care centre and its staff and children. The package contains hand puppet, shadow puppet and puppet theatre stories, conversational questions and objectives associated with the stories. The package also contains all the materials needed to create a puppet show with the kids. The goal of this thesis was to facilitate and increase the use of drama and add theatre education to the day to day education of the day care centre. Our goal was also to encourage teachers to use this material package and we hope it will also inspire children to use the puppets independently. In addition, the purpose was that the kindergarten teachers could easily create and play different kinds of puppet shows with the children. All the stories in the package were planned so that they can be utilised with children of all ages. The functional part of the thesis contained planning, designing of the material package and testing it with the staff and children of the day care centre. The functionality of the material package was assessed through the feedback of both the staff and children. After the assessment the material package will be given to the day care centre and they will be able to continue developing it if they wish to do so. The theoretical framework of this thesis concentrated on the development of early childhood education in Finland, creative methods, the new early childhood education plan and drama education from the perspective of different early childhood education plans. The functional part of the thesis contained planning, designing of the material package and testing it with the staff and children of the day care centre. The functionality of the material package was assessed through the feedback of both the staff and children. After the assessment the material package will be given to the day care centre and they will be able to continue developing it if they wish to do so

Spaceboard II Structural Panels: Forming Apparatus and Methods

Novel methods and a patented apparatus have been developed to fabricate Spaceboard II, a new unique structural board of pulped wood fiber. Like the earlier Spaceboard I, the board has flat, high-density faces and a core of rectangular cells defined by high-density cell walls formed integrally with the faces. The board is assembled from two asymmetrical panel halves joined cell to cell. The panels are formed (molded) at low bulk density and subsequently compacted to a unique shape and uniform high density. Spaceboard II is formed by the porous mandrel method, which allows fabrication of much thicker panels than was possible with the original Spaceboard I method. A variety of wet or dry (adhesive-coated) fiber furnish may be used, with either air or water as the forming vehicle. The boards are being investigated for use in light frame and commercial construction; for packaging, palletizing, partitions, and furniture; and for other uses. In the present study, a total of 55 panels, 635 by 1,244 by 38 mm thick, were made by wet-and dry-forming methods in a Forest Products Laboratory patented apparatus

SPECT image segmentation for estimation of tumour volume and activity concentration in 177Lu-DOTATATE radionuclide therapy

Background: Dosimetry in radionuclide therapy has the potential to allow for a treatment tailored to the individual patient. One therapeutic radiopharmaceutical where patient-specific dosimetry is feasible is 177Lu-DOTATATE, used for the treatment of neuroendocrine tumours. The emission of gamma photons by 177Lu allows for imaging with SPECT (single photon emission computed tomography). One important step for dosimetry using this imaging technique is the SPECT image segmentation, which needs to be robust and accurate for the estimated quantities to be reliable. This work investigates different methods for automatic tumour delineation in 177Lu-DOTATATE SPECT images. Three segmentation methods are considered: a fixed 42% threshold (FT), the Otsu method (OM) and a method based on Fourier surfaces (FS). Effects of including resolution compensation in the iterative SPECT image reconstruction are also studied. Evaluation is performed based on Monte Carlo-simulated SPECT images from 24 h and 336 h post injection (p.i.), for determination of the volume, activity concentration and dice similarity coefficient. In addition, patient data are used to investigate the correspondence of tumour volumes when delineated in SPECT or morphological CT or MR images. Patient data are also used to examine the sensitivity to the operator-dependent initialization. Results: For simulated images from 24 h p.i. reconstructed without resolution compensation, a volume and activity-concentration root-mean-square error below 15% is typically obtained for tumours above approximately 10 cm3 when using OM or FS, while FT performs considerably worse. When including resolution compensation, the tumour volume becomes underestimated and the activity concentration overestimated. The FS method appears to be robust to noise, as seen for the 336 h images. The differences between the tumour volumes estimated from the SPECT images and the volumes estimated from morphological images are generally larger than the discrepancies seen for the simulated data sets. Conclusions: Segmentation results are encouraging for future dosimetry of tumours with volumes above approximately 10 cm3. Using resolution compensation in the reconstruction may have a negative effect on volume estimation

Die Bestimmung des Nichtmetalls Fluor mit High-Resolution-Continuum Source-Molekülabsorptionsspektrometrie (HR-CS-MAS)

Es wird eine Methode zur Bestimmung des Gesamtgehaltes von Fluor in verschiedenen Matrices vorgestellt, die erstmalig unabhängig von der Bindungsform des Analyten Fluor ist und auch für nichtwässrige Lösungen eingesetzt werden kann. Die Methode beruht auf der analytischen Auswertung der Molekülabsorption (MA) von Galliummonofluorid mit einem hochauflösenden Kontinuumstrahler Atomabsorptions-spektrometer (HR CS AAS) mit quergeheiztem Graphitrohrofen. Den höchsten Absorptionskoeffizienten hat die Rotationslinie von GaF bei einer Wellenlänge von 211,248 nm. Das Elektronenanregungsspektrum wird in einem permanent mit Zirkoniumcarbid beschichteten Graphitrohr mit PIN Plattform durch die Zugabe von bis zu 500 µg Ga als Molekülbildungsreagenz erzeugt. Durch die Methode der statistischen Versuchsplanung wurde der empfindlichkeits-steigernde Einfluss verschiedener Modifier auf die GaF MA untersucht. Die höchste Methodenempfindlichkeit wurde durch die Verwendung eines Pd/ Zr- und eines Natriumacetat Modifiers erreicht. Entscheidende Voraussetzung für die Wirksamkeit des Palladiums war die Einführung einer thermischen Vorbehandlung zur Aktivierung des Pd/ Zr Modifiers. Unter diesen Bedingungen wurden eine Pyrolysetemperatur von 550° C und eine Molekülbildungstemperatur von 1550° C optimiert. Für ein Probeninjektionsvolumen von 20 µL wurde eine Nachweisgrenze von 0,26 µg L-1 F ( 5,2 pg F) und eine charakteristische Konzentration von 0,37 µg L-1 F ( 7,4 pg F) erreicht. Damit ist die Methode mehr als eine Größenordnung nachweisstärker als die ausschließlich für wässrige Lösungen einsetzbare Ionenchromatographie (IC) und die ionensensitive F Elektrode (F ISE). Die Richtigkeit der Methodenergebnisse konnte in verschiedenen Matrices wie Wasser, Zahncreme, Sediment, Pflanzen, Futtermittel nachgewiesen sowie anhand von zertifi-zierten Referenzmaterialien und durch Vergleichsmessungen mit anderen Analysen-verfahren zur Bestimmung von Fluor (F-ISE und GC MS) bestätigt werden

Estudio de la composición y el rol catalítico de metales de transición en reacciones de oxidación en vinos

122 P.El vino es un producto de gran interés comercial y científico, principalmente por consideraciones organolépticas, complejidad química y por los conocidos beneficios a la salud derivados de su consumo moderado. La calidad del vino y sus efectos biológicos dependen fundamentalmente de su composición química, la que puede variar de manera importante dependiendo del tipo de materia prima y las practicas de producción utilizadas en su elaboración. Un tema de la enología que todavía no está absolutamente resuelto corresponde al rol catalítico de los metales de transición en las reacciones de oxidación que ocurren en la matriz del vino. Lo anterior, probablemente tiene que ver con la complejidad y constantes cambios que experimenta el producto durante su producción y envejecimiento. Este estudio se centra en la caracterización de ciertos metales y de componentes orgánicos, cómo sus Concentraciones pueden ser moduladas por diferentes tratamientos de descube, evaluar la interación entre especies oxidantes y reductoras en soluciones de vino modelo mediante electroquímica, y evaluar el uso de espectrometría de masas para la caracterización y clasificación de vinos comerciales. Los principales resultados obtenidos dan cuenta de que los niveles de metales mayoritarios en vinos chilenos están dentro del los rangos esperados a nivel mundial; también, que el tiempo de descube es determinante en la concentración de metales mayoritarios. A nivel de solución modelo, fue posible observar que la oxidabilidad de los fenoles está determinada por la presencia de metales y del ion bisulfito. Finalmente, fue posible discriminar vinos chilenos por cultivar y por valle de origen analizando su perfil orgánico y/o su concentración de metales mayoritarios. En conclusión, los niveles de metales en vinos pueden ser modulados por tratamientos enológicos y tienen un efecto catalizador en la oxidación de ciertos fenoles./ABSTRACT:Abstract Wine is a food product that produces great commercial and scientific interest mainly because of its sensory characteristics, chemical complexity and also due to the well-known health benefits derived from its moderate consumption. The quality of the product and its biological effects are mainly dependent on the wine’s chemical composition, which can vary greatly depending on the type of grapes and production practices used during winemaking. One aspect of winemaking that is not fully comprehended has to do with the catalytic role of transition metals in wine oxidation reactions. The prior probably has to do with the complexity and continuous changes that the wine matrix experiments during production and aging. This study focuses on the characterization of certain metals and organic compounds, the way in which their concentrations can be modulated by different racking treatments, an lectrochemical study of the interaction between oxidizing and reducing species in model wine solutions, and an evaluation of the use of mass spectrometry on the characterization and classification of commercial wines. The main results obtained show that the concentration of major metal ions of Chilean wines are within the expected ranges observed worldwide, and that the racking time is crucial in the concentration of these ions. In model solution, it was possible to observe that phenolic oxidizability is highly dependent on the presence of metals and bisulfite ion. Finally, it was possible to discriminate Chilean wines by variety and valley of origin based on the statistical analysis of their organic profile and/or concentration of major metals.Therefore, it was possible to conclude that the levels of metals in wines can be modulated by certain oenological practices and that they have a catalytic effect on the oxidation of certain phenols

Evaluación sensorial de vinos desalcoholizados por pruebas de diferencias y ranking

36 p.Los procesos de desalcoholización de vinos pueden producirmodificaciones sensoriales que no han sido bien estudiadas. En este estudio, se realizaron dos tipos de evaluaciones sensoriales, para evaluar las diferencias y preferencias entre vinos tintos de las cepas País y Cabernet Sauvignon, desalcoholizados (0 y 6% de etanol V/V), utilizando las metodologías de osmosis inversa y conos rotatorios. Para dicho efecto se conformaron paneles sensoriales con personal y estudiantes de la universidad de Talca quienes evaluaron los vinos a través de pruebas triangulares (36 jueces) y pruebas de preferencias (17 jueces). Los resultados de los test triangulares indicaron que los panelistas fueron capaces de diferenciar significativamente todos los tratamientos; esto es: (a) los vinos de la variedad País desalcoholizados al 0 y 6% V/V de etanol mediante osmosis inversa (p<0,001), (b) los vinos de la variedad País desalcoholizados al 0 y 6% V/V de etanol mediante conos rotatorios (p<0,001), (c) los vinos de la variedad Cabernet Sauvignon desalcoholizados al 0 y 6% V/V de etanol mediante osmosis inversa (p<0,01), (d) los vinos de la variedad País desalcoholizados al 0% V/V de etanol mediante osmosis inversa y conos rotatorios (p<0,001), y (e) los vinos de la variedad País desalcoholizados al 6% V/V de etanol mediante osmosis inversa y conos rotatorios (p<0,001). Los resultados del test de preferencias donde los vinos fueron analizados separadamente para cada variedad arrojaron lo siguiente: (a) para los vinos País, se observó un favoritismo estadísticamente significativo (p<0,05) para los tratamientos de desalcoholización mediante osmosis inversa; (b) para los vinos Cabernet Sauvignon, no se obtuvieron diferencias estadísticamente significativas. Cuando todas las muestras de vinos fueron analizadas simultáneamente, el favoritismo lo obtuvo la muestra de Cabernet Sauvignon con 6% V/V etanol, tratado con osmosis inversa./ABSTRACT: The sensorial changes take place during the dealcoholization process, are an important topic that requires more attention in order to fill the gaps of information that exist in this subject. Therefore, in the context of an investigation carried out by the National Institute of Agricultural Research (INIA), entitled “Investigation, development and innovation in the production of wines with low levels of alcohol in Chile” is that we developed different sensory evaluations to estimate possible differences and consumer preferences between partially dealcoholized wines. The wines studied were made with the varieties País and Cabernet Sauvignon, dealcoholized to 0 and 6% ethanol V/V using reverse osmosis and spinning cones methodologies. The first sensory panels consisted of 36 judges (students and staff from the Universidad de Talca), that compared the wines based on alcohol content and dealcoholization method using triangular tests. Subsequently, 17 highly discriminating panelists were selected to perform preference test, grouping the samples according to varieties, and an assessment where all samples were compared simultaneously. The results of the triangle tests indicated that the panelists were able to significantly discriminate all treatments (i.e. (a) the wines of the País variety, dealcoholized to a 0 and 6% V/V ethanol by reverse osmosis (p<0,001), (b) the wines of the País variety, dealcoholized to a 0 and 6% V/V ethanol by spinning cones (p<0,001), (c) the wines of the Cabernet Sauvignon variety, dealcoholized to a 0 and 6% V/V ethanol by reverse osmosis (p<0,001), (d) the wines of the Cabernet Sauvignon variety dealcoholized to a 0 and 6% V/V ethanol by spinning cones (p<0,001), (d) the wines of the País variety dealcoholized to a 0% V/V ethanol by reverse osmosis and spinning cones (p<0,001), (e) the wines of the País variety dealcoholized to a 6% V/V ethanol by reverse osmosis and spinning cones (p<0,001). The results of the preferences test in which the wines of each variety were analyzed separately showed the following results: (a) for the País wines, the preferred wines were those dealcoholized by reverse osmosis, (b) for the Cabernet Sauvignon wines the differences were not statistically significant. When all of the wine samples were analyzed simultaneously, the Cabernet Sauvignon sample with 6% V/V ethanol, dealcoholized with reverse osmosis was the preferred sample