Peptide-mediated oligonucleotide delivery into eucaryotic cells

Nukleinsäuren sind viel versprechende Moleküle um genetisch bedingte Erkrankungen zu behandeln. Die unzureichende Serumstabilität und die schlechte Zellaufnahme aufgrund des hohen Molekulargewichts, der negativen Ladung und des hydrophilen Charakters der Nukleinsäure, erschweren jedoch die klinischen Anwendungen. Des Weiteren müssen bevor die Nukleinsäuren einem Patienten verabreicht werden können, geeignete in vitro-Systeme entwickelt werden, um die Auswirkungen der Nukleinsäure auf die Zellen zu untersuchen. Daher wurden zahlreiche virale als auch nicht-virale Transportsysteme entwickelt, um den Transport der therapeutischen Nukleinsäuren in die Zellen zu erleichtern. Da jedoch die Anwendung von viralen Vektoren mit vielen Nachteilen verbunden ist, werden nicht-virale Transportsysteme mit einem guten Wirkungsgrad und einer geringen Toxizität dringend benötigt. In den letzten Jahren sind sogenannte zellpenetrierende Peptide (cell penetrating peptides, CPP) als effektive, nicht-virale Vektoren für den Nukleinsäure-Transfer in den Fokus der Forschung getreten. Sie sind in der Lage, Moleküle in das Zellinnere zu transportieren, welche aufgrund ihrer Ladung, Größe und Hydrophilie normalerweise nicht die Zellmembran passieren können. Ziel dieser Arbeit war die Etablierung von geeigneten in vitro-Systemen sowie die Entwicklung von neuen peptid-basierten Transportmolekülen, die über eine nicht-kovalente Verbindung in der Lage sind, die Nukleinsäuren in die Zellen zu transportieren. Hierfür wurden verschiedene CPPs im Hinblick auf ihre Transportfähigkeit für Forschungsbereiche untersucht, die noch geeignete Transporter benötigen. Die untersuchten CPPs wurden sowohl von dem humanen Calcitonin (hCT) als auch von dem kationischen antimikrobiellen Peptid CAP18 (sC18) abgeleitet. Die in dieser Arbeit untersuchten CPPs waren in der Lage, Nukleinsäuren erfolgreich in verschiedene Zelllinien (MCF-7, HEK-293 und hTERT RPE-1) zu transportierten, ohne dabei die Vitalität der Zellen zu beeinflussen. Des Weiteren konnte für das CPP N-E5L-hCT(18-32)-k7 eine bemerkenswert hohe Transfektions-Effizienz erzielt werden, nachdem nicht-differenzierte hTERT RPE-1 Zellen damit transfiziert wurden. Die Effizienz überschritt sogar die von Lipofectamin, welches als positiv Kontrolle verwendet wurde. CPP N-E5L-hCT(18-32)-k7 war zudem in der Lage in einzellige Protisten (Choanoflagellate) zu internalisieren.

Montesquieu, Rousseau, Herder. De las teorías climáticas a la voz del pueblo como factor constitutivo

This article shows in which way the so-called climate theories, which have been developed since Antiquity, change over the course of time and influence the different theories on the origin of language. Via Montesquieu and Rousseau, the “climate theories” have influenced Johann Gottfried Herder, who bases on the romantic concept of Volk. By this means, a lot of ideas come into being which are fundamental for the foundation and development of the national philologies in Europe.

Conflictos entre el pueblo y las autoridades. Bubenreuth: un ejemplo actual en Baviera

The sovereign of a democratic state is „the people“. However, they transfer their voices to a few political party representatives in order to make them exercise legislative and executive powers in the name of “the people”. In different European countries, this model of representative democracy is marked by elements of direct democracy. In Switzerland, for example, there are frequent plebiscites on a number of issues and in France, the President of the Republic is elected directly. In Germany, the constitution calls for a “Volksabstimmung”, or a referendum at the federal level, a “Volksentscheid” or plebiscite at the federal state level and a “Bürgerentscheid” at the city level. But in small municipalities where everyone knows each other and people talk, a different form of direct democracy continues on. In the case of Bubenreuth, where I have lived for more than 30 years, the community dared to raise its voice against the mayor and against town councillors to have them revoke the application of a legal but unjust regulation, or for them to at least mitigate the effects.

Nombre, imagen, cuerpo, voz. Apariciones del pueblo como grupo social diferenciado en textos del siglo XIX español: proclamas de la guerra contra Napoleón, Mariano José de Larra, Rosalía de Castro

This paper analyzes the emergence of the term 'pueble', or 'people', in Spanish literature of the nineteenth century with the meaning of differentiated social subject in a corpus composed of texts of the War of Independence by Mariano José de Larra and Rosalia de Castro. The texts and authors selected are representative of their cultural and historical contexts and together they form a part of the Spanish Romanticism.

Lo que sé de los hombrecillos de Juan José Millás: la ficción psicótica de la sociedad perfecta.

This short essay deals with literary representations of identity and of social reality, especially in relation to the novelistic works published by the Spanish writer Juan José Millás. The article is divided into three parts. The first section is dedicated to a general overview of the new perspectives brought by the contemporary ‘linguistic turn’ in culture, which is currently considered as the product of different discourses and not as an ontological datum. The postmodern condition, on the other hand, is described as the age in which it has become radically difficult to rely on such ideas as “nation” and “people” for the construction of personal identity. The second part of the article identifies Millás’ poetics as an excellent example of describing the neurotic symptoms produced by the urban way of life in Western communities. Millás recognizes the separateness between language and material reality as the origin of the subject's isolation, especially in contemporary life. Finally, the third part handles with Lo que sé de los hombrecillos, the last novel by Millás, in which we witness a significant switch from neurosis to psychosis in the mind of the protagonist who offers us a distorted realisation of his idea of community and plenitude.

Un urbis encomium a lo vulgar: el caso de los dictados tópicos

Analysis of popular topics taught in the light of the topics learned poetry, revolving around the urbis encomium, understanding the popular verses somehow respond to a similar structure to that established for the praise of copyright poetry. The popular songs are close to the issue which are specific to them, collected by imitation or by spontaneous coincidence and adopt many of the tools of author's lyric poetry. Within this framework, we go deeply into analogies we found when studying the group of songs we label " songs to the earth" with the topics of classical rhetoric making up the urbis encomium, the praise for the city.

Historia en voz baja: la oralidad y el rumor para la narración de la Guerra Civil española en la ‘Primera derrota’ de Alberto Méndez

The first of the four paths that structure the book "Los girasoles ciegos", by Alberto Méndez – with its theory about the delayed end of the Spanish Civil War as a longing for destruction outside of any kind of strategy based on military logic – presents the construction of a memory based on certain oral marks, facts provided quietly by apparently non-central characters, the distrust of written documents, and the use of speech patterns mostly associated with spontaneity in order to set up a level of verisimilitude which makes the memory emerge in parallel pathways considered relatively reliable (for example, the case of a report), forged speech on the basis of indirect references, testimonials and letters. The aim of the paper is to consider an example of contemporary Spanish narrative in which a journey, perhaps weak in terms of the material, support to the channels through which the narrator comes in the story through the voice of the people – but functional as an approach to a search of the recent past – contributes to a certain conception of memory.

Asymptotic Change-Point Analysis of the Dependencies in Time Series

In recent papers, Wied and his coauthors have introduced change-point procedures to detect and estimate structural breaks in the correlation between time series. To prove the asymptotic distribution of the test statistic and stopping time as well as the change-point estimation rate, they use an extended functional Delta method and assume nearly constant expectations and variances of the time series. In this thesis, we allow asymptotically infinitely many structural breaks in the means and variances of the time series. For this setting, we present test statistics and stopping times which are used to determine whether or not the correlation between two time series is and stays constant, respectively. Additionally, we consider estimates for change-points in the correlations. The employed nonparametric statistics depend on the means and variances. These (nuisance) parameters are replaced by estimates in the course of this thesis. We avoid assuming a fixed form of these estimates but rather we use "blackbox" estimates, i.e. we derive results under assumptions that these estimates fulfill. These results are supplement with examples. This thesis is organized in seven sections. In Section 1, we motivate the issue and present the mathematical model. In Section 2, we consider a posteriori and sequential testing procedures, and investigate convergence rates for change-point estimation, always assuming that the means and the variances of the time series are known. In the following sections, the assumptions of known means and variances are relaxed. In Section 3, we present the assumptions for the mean and variance estimates that we will use for the mean in Section 4, for the variance in Section 5, and for both parameters in Section 6. Finally, in Section 7, a simulation study illustrates the finite sample behaviors of some testing procedures and estimates.

A Grammar of Tajio. A Language Spoken in Central Sulawesi

This work is a description of Tajio, a Western Malayo-Polynesian language spoken in Central Sulawesi, Indonesia. It covers the essential aspects of Tajio grammar without being exhaustive. Tajio has a medium sized phoneme inventory consisting of twenty consonants and five vowels. The language does not have lexical (word) stress; rather, it has a phrasal accent. This phrasal accent regularly occurs on the penultimate syllable of an intonational phrase, rendering this syllable auditorily prominent through a pitch rise. Possible syllable structures in Tajio are (C)V(C). CVN structures are allowed as closed syllables, but CVN syllables in word-medial position are not frequent. As in other languages in the area, the only sequence of consonants allowed in native Tajio words are sequences of nasals followed by a homorganic obstruent. The homorganic nasal-obstruent sequences found in Tajio can occur word-initially and word-medially but never in word-final position. As in many Austronesian languages, word class classification in Tajio is not straightforward. The classification of words in Tajio must be carried out on two levels: the morphosyntactic level and the lexical level. The open word classes in Tajio consist of nouns and verbs. Verbs are further divided into intransitive verbs (dynamic intransitive verbs and statives) and dynamic transitive verbs. Based on their morphological potential, lexical roots in Tajio fall into three classes: single-class roots, dual-class roots and multi-class roots. There are two basic transitive constructions in Tajio: Actor Voice and Undergoer Voice, where the actor or undergoer argument respectively serves as subjects. It shares many characteristics with symmetrical voice languages, yet it is not fully symmetric, as arguments in AV and UV are not equally marked. Neither subjects nor objects are marked in AV constructions. In UV constructions, however, subjects are unmarked while objects are marked either by prefixation or clitization. Evidence from relativization, control and raising constructions supports the analysis that AV and UV are in fact transitive, with subject arguments and object arguments behaving alike in both voices. Only the subject can be relativized, controlled, raised or function as the implicit subject of subjectless adverbial clauses. In contrast, the objects of AV and UV constructions do not exhibit these features. Tajio is a predominantly head-marking language with basic A-V-O constituent order. V and O form a constituent, and the subject can either precede or follow this complex. Thus, basic word order is S-V-O or V-O-S. Subject, as well as non-subject arguments, may be omitted when contextually specified. Verbs are marked for voice and mood, the latter of which is is obligatory. The two values distinguished are realis and non-realis. Depending on the type of predicate involved in clause formation, three clause types can be distinguished: verbal clauses, existential clauses and non-verbal clauses. Tajio has a small number of multi-verbal structures that appear to qualify as serial verb constructions. SVCs in Tajio always include a motion verb or a directional.

Atmospheric calibration for sub-millimeter radio astronomy

This thesis is focused on improving the calibration accuracy of sub-millimeter astronomical observations. The wavelength range covered by observational radio astronomy has been extended to sub-millimeter and far infrared with the advancement of receiver technology in recent years. Sub-millimeter observations carried out with airborne and ground-based telescopes typically suffer from 10% to 90% attenuation of the astronomical source signals by the terrestrial atmosphere. The amount of attenuation can be derived from the measured brightness of the atmospheric emission. In order to do this, the knowledge of the atmospheric temperature and chemical composition, as well as the frequency-dependent optical depth at each place along the line of sight is required. The altitude-dependent air temperature and composition are estimated using a parametrized static atmospheric model, which is described in Chapter 2, because direct measurements are technically and financially infeasible. The frequency dependent optical depth of the atmosphere is computed with a radiative transfer model based on the theories of quantum mechanics and, in addition, some empirical formulae. The choice, application, and improvement of third party radiative transfer models are discussed in Chapter 3. The application of the calibration procedure, which is described in Chapter 4, to the astronomical data observed with the SubMillimeter Array Receiver for Two Frequencies (SMART), and the German REceiver for Astronomy at Terahertz Frequencies (GREAT), is presented in Chapters 5 and 6. The brightnesses of atmospheric emission were fitted consistently to the simultaneous multi-band observation data from GREAT at 1.2 ∼ 1.4 and 1.8 ∼ 1.9 THz with a single set of parameters of the static atmospheric model. On the other hand, the cause of the inconsistency between the model parameters fitted from the 490 and 810 GHz data of SMART is found to be the lack of calibration of the effective cold load temperature. Besides the correctness of atmospheric modeling, the stability of the receiver is also important to achieving optimal calibration accuracy. The stabilities of SMART and GREAT are analyzed with a special calibration procedure, namely the “load calibration". The effects of the drift and fluctuation of the receiver gain and noise temperature on calibration accuracy are discussed in Chapters 5 and 6. Alternative observing strategies are proposed to combat receiver instability. The methods and conclusions presented in this thesis are applicable to the atmospheric calibration of sub-millimeter astronomical observations up to at least 4.7 THz (the H channel frequency of GREAT) for observations carried out from ∼ 4 to 14 km altitude. The procedures for receiver gain calibration and stability test are applicable to other instruments using the same calibration approach as that for SMART and GREAT. The structure of the high performance, modular, and extensible calibration program used and further developed for this thesis work is presented in the Appendix C.