Detergent-free membrane protein purification

Membrane proteins are localised within a lipid bilayer; in order to purify them for functional and structural studies the first step must involve solubilising or extracting the protein from these lipids. To date this has been achieved using detergents which disrupt the bilayer and bind to the protein in the transmembrane region. However finding conditions for optimal extraction, without destabilising protein structure is time consuming and expensive. Here we present a recently-developed method using a styrene maleic acid (SMA) co-polymer instead of detergents. The SMA co-polymer extracts membrane proteins in a small disc of lipid bilayer which can be used for affinity chromatography purification, thus enabling the purification of membrane proteins while maintaining their native lipid bilayer environment.

Musical commonplace book : manuscript, [approximately 1790-approximately 1810]

Contains songs, partly from English operas, and instrumental music.

Membrane protein production in the Yeast, S. cerevisiae

The first crystal structures of recombinant mammalian membrane proteins were solved in 2005 using protein that had been produced in yeast cells. One of these, the rabbit Ca2+-ATPase SERCA1a, was synthesized in Saccharomyces cerevisiae. All host systems have their specific advantages and disadvantages, but yeast has remained a consistently popular choice in the eukaryotic membrane protein field because it is quick, easy and cheap to culture, whilst being able to post-translationally process eukaryotic membrane proteins. Very recent structures of recombinant membrane proteins produced in S. cerevisiae include those of the Arabidopsis thaliana NRT1.1 nitrate transporter and the fungal plant pathogen lipid scramblase, TMEM16. This chapter provides an overview of the methodological approaches underpinning these successes.

PCR-Based Analysis of Mitochondrial DNA Copy Number, Mitochondrial DNA Damage, and Nuclear DNA Damage.

Because of the role that DNA damage and depletion play in human disease, it is important to develop and improve tools to assess these endpoints. This unit describes PCR-based methods to measure nuclear and mitochondrial DNA damage and copy number. Long amplicon quantitative polymerase chain reaction (LA-QPCR) is used to detect DNA damage by measuring the number of polymerase-inhibiting lesions present based on the amount of PCR amplification; real-time PCR (RT-PCR) is used to calculate genome content. In this unit, we provide step-by-step instructions to perform these assays in Homo sapiens, Mus musculus, Rattus norvegicus, Caenorhabditis elegans, Drosophila melanogaster, Danio rerio, Oryzias latipes, Fundulus grandis, and Fundulus heteroclitus, and discuss the advantages and disadvantages of these assays.

The relationship between vegetation and modern pollen assemblages on Mount Paggeo (NE Greece)

In this study, we investigated the relationship between vegetation and modern-pollen rain along the elevational gradient of Mount Paggeo. We apply multivariate data analysis to assess the relationship between vegetation and modern-pollen rain and quantify the representativeness of forest zones. This study represents the first statistical analysis of pollen-vegetation relationship along an elevational gradient in Greece. Hence, this paper improves confidence in interpretation of palynological records from north-eastern Greece and may refine past climate reconstructions for a more accurate comparison of data and modelling. Numerical classification and ordination were performed on pollen data to assess differences among plant communities that beech (Fagus sylvatica) dominates or co-dominates. The results show a strong relationship between altitude, arboreal cover, human impact and variations in pollen and nonpollen palynomorph taxa percentages.

Forest typology of broadleaf forests from Sierra Maestra, Eastern Cuba

A phytosociological study of the forests from Sierra Maestra is conducted, following the methodology of the Zurich- Montpelier School. They are transformed into a forest typology using the standards of the Institute of Agro-Forestry Research. In general, 35 types and/or subtypes are presented. From this group, the most abundant ones belong to semi-deciduous microphyll forest, followed by those from mangroves and mountain rainforest, respectively. Silvicultural treatments are needed; among them, the protection forests are those found above 800 m asl and mangroves. 

New contributions to the native and alien flora in riparian habitats of the Cantabrian watershed (Northern Spain)

As a result of a floristic survey carried out in riparian habitats of northern Spain, new chorological data are provided for 9 alien and 6 native plant species. Some species are reported for the first time at regional scale, such as Carex strigosa, Helianthus x laetiflorus and Persicaria pensylvanica in Cantabria. Also noteworthy is the finding of naturalised populations of the North American grass Muhlenbergia schreberi at the Urumea river basin, which represents the second reference for the Iberian Peninsula.

Narratives Around Iberian Culture at Valencia Museums and Archaeological Sites: From Official Discourses to Social Perceptions

Museums and archaeological sites are considered the most authoritative places for talking about the past and the heritage from a scientific perspective. In fact, visitors assume their discourses as reliable and indisputable. In spite of that, professionals of archaeology must critically analyse the production of narratives at heritage sites, since they often reflect social, political and identity issues related to the present-day realities. The aim of this paper is to study official and popular discourses about the Iberian culture (Iron Age) collected in museums and archaeological sites from Valencia region.

Prediction of Phosphorylation Sites from Protein Sequences

Phosphorylation is amongst the most crucial and well-studied post-translational modifications. It is involved in multiple cellular processes which makes phosphorylation prediction vital for understanding protein functions. However, wet-lab techniques are labour and time intensive. Thus, computational tools are required for efficiency. This project aims to provide a novel way to predict phosphorylation sites from protein sequences by adding flexibility and Sezerman Grouping amino acid similarity measure to previous methods, as discovering new protein sequences happens at a greater rate than determining protein structures. The predictor – NOPAY - relies on Support Vector Machines (SVMs) for classification. The features include amino acid encoding, amino acid grouping, predicted secondary structure, predicted protein disorder, predicted protein flexibility, solvent accessibility, hydrophobicity and volume. As a result, we have managed to improve phosphorylation prediction accuracy for Homo sapiens by 3% and 6.1% for Mus musculus. Sensitivity at 99% specificity was also increased by 6% for Homo sapiens and for Mus musculus by 5% on independent test sets. In this study, we have managed to increase phosphorylation prediction accuracy for Homo sapiens and Mus musculus. When there is enough data, future versions of the software may also be able to predict other organisms.

Sähkötoimisten käsiproteesien nykyteknologian taso

Työssä selvitettiin sähkötoimisten käsiproteesien nykyteknologian taso. Selvitettäviä asioita olivat keskushermoston ja proteesin välisen hermokytkennän toteutustapa, sähkötoimisten käsiproteesien keskeiset tekniset ominaisuudet sekä käsiproteesin mekaaniset toteutustavat. Tutkimus suoritettiin kirjallisuustutkimuksena. Työhön valittiin esimerkkejä kaupallisesti saatavilla olevista käsiproteeseista jotka löytyivät internetistä hakemalla kaikkein edisty-neintä käsiproteesia. Työstä rajattiin pois proteesin suora kytkeminen keskushermostoon. Liikeinformaation välitys aivoilta proteesille onnistuu mittaamalla lihassähkökäyrä erilaisilla ihon ja lihasten päälle, ihon alle lihaksien yhteyteen tai suoraan hermojen yhteyteen asete-tuilla elektrodeilla. Lihassähkökäyrän mittaamisessa ihon pinnalta on ongelmana sähkömag-neettinen säteily, hiki, joka muuttaa ihon impedanssia ja elektrodien meneminen pois paikal-taan. Ihon alle asetettavat elektrodit kapseloituvat, mikä heikentää niiden toimintaa ja nii-den ihon läpi kulkevat johdot voivat altistaa kohdan infektioille tai takertua johonkin. Her-moihin suorassa kosketuksissa olevat elektrodit aiheuttavat lisäksi hermopinteen. Kohden-netulla uudelleenhermotuksella voidaan hermosyyt johtaa lihaksiin, jolloin lihaksista saa-daan biologiset vahvistimet lihassähkökäyrää varten tai korvaavalle ihoalueelle, johon koh-distuva kosketus tuottaa tuntemuksen käteen kohdistuvasta kosketuksesta. Käden menet-tämisen myötä menetettävät hermo-ohjaustiedot voivat osittain korvautua aivojen mukau-tuvuuden ansiosta, mikä mahdollistaa tekokäden käyttämisen oppimisen samalla tavoin kuin polkupyörällä ajon. Hermotakaisinkytkentä mahdollistaa proteesin paremman hallin-nan. On mahdollista valmistaa keinoihoa johon kohdistuva paine saa aikaan muutoksen sen sähköisissä ominaisuuksissa, mitä voidaan sitten käyttää varsinaisen hermoärsytyksen luo-van laitteen, kuten tynkää ärsyttävän täryttimen, ohjaamisessa. On mahdollista valmistaa keinolihaksia joiden avulla nivelten liike voidaan toteuttaa luonnollisen kaltaisilla rakenteilla ja jotka ovat jopa kymmeniä kertoja voimakkaampia kuin aidot lihakset. Nykyteknologian avulla on mahdollista rakentaa käsiproteesi joka liikeradoiltaan, voimal-taan ja hermotakaisinkytkennän osalta vastaa lähes täydellisesti aitoa ihmiskättä. Haasteena on vielä kokeiluasteella oleva teknologian taso sekä korkea hinta.

Programa ciclovida como política de mobilidade urbana sustentável: estudo empírico na Universidade Federal do Paraná

The urban mobility has in the bicycle its symbol of the most environmentally sustainable modal of transportation of the planet, title conferred by the United Nations Organization (ONU). Nevertheless, the cities suffer with the traffic jam caused by the automobile fleet growth, what implies in a larger displacement time, injuries to the environment, to financials and to the quality of life. In order to build in a sustainable way the urban mobility, public policies are necessary to encourage the preferential usage, by the society, of the mass transit or the non-motorized type of transportation. Objecting cooperate with the public power and the civil society, this study intents to, through an analysis about the UFPR Extension Program - CICLOVIDA, propose a sustainable urban mobility public policy development, with emphasis in the bicycle´s usage. eferentially, it is discussed concepts such as Society, State and Government as well as public policies, which go in cycles to the State´s and Government´s responsibilities; and are presented the Sustainable Urban Mobility Public Policies (PPMUS) in Brazil. Also, it is debated about the transport by bicycle as an option to the sustainable mobility. Methodologically, the study is of an empiric-social nature, with the application of both qualitative and observational methods, what characterizes as an exploratory level of research, with the researcher´s involvement in the participant form, due to her acting on the Program. Regarding the research lineation, it was used the bibliographic and documental form as well as case study, by means of the nonparametric sampling by accessibility or convenience. Related to the data collection technique, it was used structured interviews with self-applied questionnaires and natural participant observation. Aligned with the research´s objective and matter, the outcome confirmed that the Extension Program CICLOVIDA of the Federal University of Paraná (UFPR) may effectively contribute in the development of a PPMUS proposal with emphasis in the bicycle usage. Thus, it was built a political proposal with a free form writing, objecting subsidize policies both institutional as public MUS through the bicycle usage and therefore, support the mobility, also contributing to the sustainability and to the public development planning.

Antonio Salieri's La calamita de' cuori (1774): Sources, Form, Context

Antonio Salieri’s La calamita de’ cuori (1774) warrants musicological attention for what it can tell us about Salieri’s compositional craft and what it reveals about the development of form in Viennese Italian-language comic opera of the mid- and late-eighteenth century. In Part I of this dissertation, I explore the performance history of La calamita, present the first plot synopsis and English translation of the libretto, and describe the variants between Carlo Goldoni’s 1752 libretto and the revised version created for Salieri’s opera. I have collated Salieri’s holograph score, Österreichische Nationalbibliothek, Vienna, Mus. Hs. 16.508, with four copies having different relationships to it, and I propose a stemma that represents the relationships between these five sources. The analyses in Part II contribute to our understanding of formal practices in eighteenth-century drammi giocosi. My study of Salieri’s La calamita reveals his reliance on a clearly defined binary structure, referred to in this dissertation as “operatic binary form,” in almost half of the arias, ensembles, and instrumental movements of this opera. Salieri’s consistent use of operatic binary form led me to explore its use in drammi giocosi by other prominent composers of this time, including Baldassare Galuppi’s La calamita de’ cuori (1752), Wolfgang Amadeus Mozart’s Il dissoluto punito, ossia Il Don Giovanni (1787), and selected arias by Pasquale Anfossi, Florian Leopold Gassmann, Giuseppe Gazzaniga, Franz Joseph Haydn, Giovanni Paisiello, and Niccolò Piccinni dating from 1760 to 1774. This study showed that Salieri and his peers adhered to a recognizable tonal plan and set of design elements in their operatic binary forms, and that their arias fall into three distinct categories defined by the tonality at the beginning of the second half of the binary structure. The analysis presented here adds to our present understanding of operatic form in mid- and late-century drammi giocosi and shows that in La calamita de’ cuori, Salieri was following the normative formal procedures of his time.

Entwicklung eines empirisch überprüfbaren Modells musikpraktischer Kompetenz (KOPRA-M)

Nachdem für das Schulfach Musik mit dem Kompetenzmodell „Musik wahrnehmen und kontextualisieren" ein erstes empirisch validiertes Kompetenzmodell vorliegt (Jordan et al., 2012), soll mit dieser Studie in Form eines theoriebasierten Modells musikpraktischer Kompetenz der erste Schritt zur empirischen Modellierung eines weiteren Bereichs, nämlich des musikpraktischen, vorgenommen werden. (DIPF/Orig.)

Botulinum toxin in masticatory muscles of the adult rat induces bone loss at the condyle and alveolar regions of the mandible associated with a bone proliferation at a muscle enthesis

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