Status quo and developments of pre-implantation genetic diagnosis. TAB-Fokus

Pre-implantation genetic diagnosis enables the genetic testing of embryos in the process of in vitro fertilisation before they are transplanted. The method raises ethical questions, affects different societal interests and therefore represents a continuing legislative challenge. In 2011, balancing individual and societal interests with regard to the handling of embryos and discrimination against those affected, a majority of the German Bundestag voted for a strictly limited authorisation. To date, with around 300 applications per year, the number of people making use of PGD corresponds to the estimations formulated in the legislative process. The further development depends on questions regarding reimbursement, the number of indications approved by the PGD Ethics Committees and the clarification of open legal questions

Etablierung von Hochdurchsatz-Kultivierungs- und -Screeningmethoden für phototrophe Einzeller

Mit Hochdurchsatz-Kultivierungs- und -Screeningmethoden können viele Proben parallel, miniaturisiert und kostengünstig bearbeitet werden. Für phototrophe Organismen wie Mikroalgen und Cyanobakterien sind Hochdurchsatz-Kultivierungsverfahren jedoch bis heute kaum etabliert. Im Rahmen dieser Arbeit wurden diese Verfahren beispielhaft für das Cyanobakterium Synechocystis sp. PCC 6803 etabliert. Die benötigte technische Automatisierung wurde hierbei durch den Einsatz eines Tecan Genesis RSP 150 Pipettierroboters erreicht. Die Kultivierung erfolgte in Deepwell-Mikrotiterplatten innerhalb einer speziell angefertigten Kammer mit programmierbaren Schüttlern, einstellbarer Belichtung und CO2-Atmosphäre. Die in diesem System erreichten Wachstumsraten sind vergleichbar mit publizierten Kultivierungsmethoden. Das Hochdurchsatz-Screening wurde mit Hilfe eines in den Roboter integrierten Tecan Genios Plus Plattenreaders durchgeführt. Es wurden beispielhaft Methoden zur Bestimmung von optischer Dichte und Chlorophyllgehalt etabliert. Die hier vorgestellte Plattform kann vielseitig zur Analyse phototropher Organismen eingesetzt werden und ist durch entsprechende Assays leicht zur Messung anderer Parameter erweiterbar.High-throughput cultivation and screening methods allow a parallel, miniaturized and cost efficient processing of many samples. These methods however, have not been generally established for phototrophic organisms such as microalgae or cyanobacteria. In this work we describe and test high-throughput methods with the model organism Synechocystis sp. PC6803. The required technical automation for these processes was archived with a Tecan Genesis RSP 150 pipetting robot. The cultivation was performed in deepwell microtiter plates within a specially constructed cultivation chamber. The chamber is outfitted with programmable shaking conditions, variable illumination and an adjustable CO2 atmosphere. The growth rates archived within this system are comparable to those achieved with established methods such as bioreactors. The high-throughput screening was achieved with a Tecan Genios Plus plate reader integrated within the pipetting robot. Methods for determination of optical density and amount of chlorophyll were established within the scope of this work. The presented platform can be used for a variety of analyses of phototrophic organisms and is easily expandable with further assays to screen for additional targets

Mémoire épique et Génie du lieu, études réunies par Caroline Cazanave

« La ‘fleur des histoires épiques’ est d’une espèce qui se révèle immédiatement voyageuse ». Ainsi débute le deuxième hors série de la collection « Bien Dire et Bien Aprandre » du Centre d’études médiévales et dialectales de Lille 3, dédié à Marie-Madeleine Castellani, et dont la thématique est celle de la translatio épique. Plus exactement, le présent volume s’attache à montrer, de manière fort convaincante, à travers une importante étude introductive et une dizaine d’articles réunis par Car..

L’art et la culture visuelle du shi’isme duodécimainde l’époque safavide à nos jours

L’histoire du shi’isme duodécimain en Iran est un centre d’intérêt des chercheurs depuis des décennies. Les textes prémodernes de théologie shi’ite, en particulier la littérature de Hadith, les œuvres exégétiques et les chroniques historiques, exposent, entre autres, les relations complexes et variables du shi’isme avec le pouvoir politique. Cependant, les formulations diverses de la doctrine shi’ite n’ont jamais été restreintes au seul domaine textuel. Depuis la période ilkhanide jusqu’à nos jours, et surtout pendant l’époque safavide lorsque le shi’isme a été proclamé, la religion officielle en Iran, sa représentation en images peintes démontre qu’il a bénéficié d’une vie visuelle remarquablement riche en Iran grâce notamment au mécénat des productions de prestige[…

Jean-Pierre Martin, Temps, mémoire, narration (Discours de l’épopée médiévale 2), 2020

La narratologie moderne dispose de nombreux outils permettant aux lecteurs et aux chercheurs d’interroger la temporalité du récit ; on pensera par exemple à la distinction entre temps de l’énoncé et temps de l’énonciation héritée de Benveniste, aux réflexions de Gérard Genette sur les « discours du récit » ou encore à la somme philosophique de Paul Ricoeur, Temps et récit. Les ouvrages théoriques consacrés à la temporalité dans la littérature médiévale sont plus rares, si l’on excepte l’œuvre..

Interaction between subunit C (Vma5p) of the yeast vacuolar ATPase and the stalk of the C-depleted V-1 ATPase from Manduca sexta midgut

AbstractProjection maps of a V1–Vma5p hybrid complex, composed of subunit C (Vma5p) of Saccharomyces cerevisiae V-ATPase and the C-depleted V1 from Manduca sexta, were determined from single particle electron microscopy. V1–Vma5p consists of a headpiece and an elongated wedgelike stalk with a 2.1×3.0 nm protuberance and a 9.5×7.5 globular domain, interpreted to include Vma5p. The interaction face of Vma5p in V1 was explored by chemical modification experiments

Structural Determination of Functional Units of the Nucleotide Binding Domain (NBD94) of the Reticulocyte Binding Protein Py235 of Plasmodium yoelii

Invasion of the red blood cells (RBC) by the merozoite of malaria parasites involves a large number of receptor ligand interactions. The reticulocyte binding protein homologue family (RH) plays an important role in erythrocyte recognition as well as virulence. Recently, it has been shown that members of RH in addition to receptor binding may also have a role as ATP/ADP sensor. A 94 kDa region named Nucleotide-Binding Domain 94 (NBD94) of Plasmodium yoelii YM, representative of the putative nucleotide binding region of RH, has been demonstrated to bind ATP and ADP selectively. Binding of ATP or ADP induced nucleotide-dependent structural changes in the C-terminal hinge-region of NBD94, and directly impacted on the RBC binding ability of RH.In order to find the smallest structural unit, able to bind nucleotides, and its coupling module, the hinge region, three truncated domains of NBD94 have been generated, termed NBD94(444-547), NBD94(566-663) and NBD94(674-793), respectively. Using fluorescence correlation spectroscopy NBD94(444-547) has been identified to form the smallest nucleotide binding segment, sensitive for ATP and ADP, which became inhibited by 4-Chloro-7-nitrobenzofurazan. The shape of NBD94(444-547) in solution was calculated from small-angle X-ray scattering data, revealing an elongated molecule, comprised of two globular domains, connected by a spiral segment of about 73.1 A in length. The high quality of the constructs, forming the hinge-region, NBD94(566-663) and NBD94(674-793) enabled to determine the first crystallographic and solution structure, respectively. The crystal structure of NBD94(566-663) consists of two helices with 97.8 A and 48.6 A in length, linked by a loop. By comparison, the low resolution structure of NBD94(674-793) in solution represents a chair-like shape with three architectural segments.These structures give the first insight into how nucleotide binding impacts on the overall structure of RH and demonstrates the potential use of this region as a novel drug target

An improved purification of ECF1 and ECF1F0 by using a cytochrome bo-deficient strain of Escherichia coli facilitates crystallization of these complexes

AbstractA novel strategy, which employs a cytochrome bo-lacking strain (GO104) and a modified isolation procedure provides an effective approach for obtaining much purer preparations of ECF1F0 than described previously, as well as for isolating homogeneous and protein-chemically pure ECF1. ECF1 obtained in this way could be crystallized by vapor-diffusion using polyethylene glycol (PEG) as a precipitant in a form suitable for X-ray diffraction analysis. The crystals belong to the orthorhombic space group P212121, with lattice parameters a=110, b=134, and c=269 Å, and diffract to a resolution of at least 6.4 Å