Development of a species-specific RNA polymerase I-based shRNA expression vector

RNA interference (RNAi) can be induced in vitro either by application of synthetic short interfering RNAs (siRNAs), or by intracellular expression of siRNAs or short hairpin RNAs (shRNAs) from transfected vectors. The most widely used promoters for siRNA/shRNA expression are based on polymerase III (Pol III)-dependent transcription. We developed an alternative vector for siRNA/shRNA expression, using a mouse RNA polymerase I (Pol I) promoter. Pol I-dependent transcription serves in cells for production of ribosomal RNA (rRNA), and as such, is ubiquitously and stably active in different cell types. As Pol I-dependent transcription is highly species-specific, Pol I-based system provides an important biosafety advantage with respect to silencing of genes with unknown functions

Entangled Stories: The Red Jews in Premodern Yiddish and German Apocalyptic Lore

“Far, far away from our areas, somewhere beyond the Mountains of Darkness, on the other side of the Sambatyon River…there lives a nation known as the Red Jews.” The Red Jews are best known from classic Yiddish writing, most notably from Mendele's Kitser masoes Binyomin hashlishi (The Brief Travels of Benjamin the Third). This novel, first published in 1878, represents the initial appearance of the Red Jews in modern Yiddish literature. This comical travelogue describes the adventures of Benjamin, who sets off in search of the legendary Red Jews. But who are these Red Jews or, in Yiddish, di royte yidelekh? The term denotes the Ten Lost Tribes of Israel, the ten tribes that in biblical times had composed the Northern Kingdom of Israel until they were exiled by the Assyrians in the eighth century BCE. Over time, the myth of their return emerged, and they were said to live in an uncharted location beyond the mysterious Sambatyon River, where they would remain until the Messiah's arrival at the end of time, when they would rejoin the rest of the Jewish people. This article is part of a broader study of the Red Jews in Jewish popular culture from the Middle Ages through modernity. It is partially based on a chapter from my book, Umstrittene Erlöser: Politik, Ideologie und jüdisch-christlicher Messianismus in Deutschland, 1500–1600 (Göttingen: Vandenhoeck & Ruprecht, 2011). Several postdoctoral fellowships have generously supported my research on the Red Jews: a Dr. Meyer-Struckmann-Fellowship of the German Academic Foundation, a Harry Starr Fellowship in Judaica/Alan M. Stroock Fellowship for Advanced Research in Judaica at Harvard University, a research fellowship from the Heinrich Hertz-Foundation, and a YIVO Dina Abramowicz Emerging Scholar Fellowship. I thank the organizers of and participants in the colloquia and conferences where I have presented this material in various forms as well as the editors and anonymous reviewers of AJS Review for their valuable comments and suggestions. I am especially grateful to Jeremy Dauber and Elisheva Carlebach of the Institute for Israel and Jewish Studies at Columbia University, where I was a Visiting Scholar in the fall of 2009, for their generous encouragement to write this article. Sue Oren considerably improved my English. The style employed for Romanization of Yiddish follows YIVO's transliteration standards. Unless otherwise noted, translations from the Yiddish, Hebrew, German, and Latin are my own. Quotations from the Bible follow the JPS translation, and those from the Babylonian Talmud are according to the Hebrew-English edition of the Soncino Talmud by Isidore Epstein

γ-Synucleinopathy: neurodegeneration associated with overexpression of the mouse protein

The role of α-synuclein in pathogenesis of familial and idiopathic forms of Parkinson’s disease, and other human disorders known as α-synucleinopathies, is well established. In contrast, the involvement of two other members of the synuclein family, β-synuclein and γ-synuclein, in the development and progression of neurodegeneration is poorly studied. However, there is a growing body of evidence that α-synuclein and β-synuclein have opposite neuropathophysiological effects. Unlike α-synuclein, overexpressed β-synuclein does not cause pathological changes in the nervous system of transgenic mice and even ameliorates the pathology caused by overexpressed α-synuclein. To assess the consequences of excess expression of the third family member, γ-synuclein, on the nervous system we generated transgenic mice expressing high levels of mouse γ-synuclein under control of Thy-1 promoter. These animals develop severe age- and transgene dose-dependent neuropathology, motor deficits and die prematurely. Histopathological changes include aggregation of γ-synuclein, accumulation of various inclusions in neuronal cell bodies and processes, and astrogliosis. These changes are seen throughout the nervous system but are most prominent in the spinal cord where they lead to loss of spinal motor neurons. Our data suggest that down-regulation of small heat shock protein HSPB1 and disintegration of neurofilament network play a role in motor neurons dysfunction and death. These findings demonstrate that γ-synuclein can be involved in neuropathophysiological changes and the death of susceptible neurons suggesting the necessity of further investigations of the potential role of this synuclein in disease

Identification of genes differentially expressed upon cocaine treatment and of new rat semaphorins

Au niveau comportmental la dépendance est définie comme un désir compulsif de prendre de la drogue avec perte de controle d’actes apparemment volontaires comme la recherche et la consommation de stupéfiants. Au niveau neuronal les stupéfiants provoquent deux types de réponses: des adaptations neuronales, c'est-à-dire des réponses homeostatiques à une stimulation excessive, et la plasticité synaptique. Au niveau moleculaire ces deux effets sont liés à des changements simultanés du mode d'expression de gènes dans le système dopaminergique mésolimbique. Les gènes différentiellement exprimés après traitement aux drogues, comprennent des facteurs de la transcription, des neurotransmetteurs, et des molecules de guidance axonale. Une image détaillée des changements d'expression genique est nécessaire pour disséquer le rôle des différentes molécules dans le développement des diverses phases temporelles et fonctionnelles associées à la dépendance. C'est pourquoi, dans l'étude présente, on a effectué la recherche des gènes différentiellement exprimés après traitement aigu à la cocaine et après retrait, dans trois régions du cerveau: le striatum, le tegmentum et l'hippocampe. Deux modifications de la méthode de display différentiel de mARN ont été introduites, permettant, premièrement, d'augmenter la reproductibilité et de faciliter le maniement de la méthode, et deuxièmement, d'effectuer une recherche spécifique de sémaphorines, une famille des molécules de guidance axonale. Les gènes exprimés différentiellement representent une large palette de candidats aux charactéristiques fonctionnelles diverses, illustrant la complexité et la variabilité des changements neuroplastiques induits par un traitement à un psychostimulant dans le cerveau. En particulier, nous avouns trouvé que le CD81, une tetraspanine, est surexprimé dans le noyau accumbens après treatment à la cocaine. Une étude par hybridisation in situ a permis de localizer le mARN de CD81 dans des régions du cerveau fonctionnellement liées à la régulation de la fonction cardiovasculaire et de l'homéostase des fluides. La plupart de ces régions expriment aussi le neuropeptide galanine. En plus, les souris déficientes pour le gène du CD81, démontraient des changements de la sensibilité à la cocaine, variables selon le sexe, ainsi que des modifications du métabolisme de la dopamine. En outre, nous avons montré qu’une proteine, liée aux processus neurodegeneratifs dans le cerveau, la g-synucleine, était sur-exprimée dans le tegmentum après traitement à la cocaine, ce qui suggère un lien entre les changements neuroplastiques liées à la neurodégénérescence et la dépendance. Finalement, un nouveau membre de la famille des sémaphorines de vertebrés et cinq orthologues de sémaphorines des classes 3,4 et 7 chez le rat, ont été identifiés, alors que l'application de la nouvelle méthode du Display Différentiel Enrichi que nous avons developpé a permis de démontrer la régulation différentielle des sémaphorines dans diverses régions du cerveau après traitement à la cocaine.At the behavioral level drug addiction is defined as compulsion to take a drug with loss of control over apparently voluntary acts of drug seeking and drug taking. At the neuronal level drugs elicit two types of responses: neuronal adaptations, i.e. homeostatic responses to excessive stimulation, and synaptic plasticity. At the molecular level both processes are paralleled by changes in gene expression pattern in the mesolimbic dopaminergic system. The genes differentially expressed upon drug treatment include transcription factors, neurotransmitter and axon guidance cues. The detailed picture of gene expression changes is necessary in order to dissect the role of different molecules in the development of the distinct temporal and functional phases of drug addiction. Therefore, in the present study the screening for differentially expressed genes upon acute cocaine withdrawal was performed in three brain regions: the striatum, the tegmentum and the hippocampus. Two modifications of differential display of mRNA were introduced allowing first for increased reproducibility and easy handling of the method, and second for a gene-specific screening for the semaphorins, a family of axon guidance cues. The differentially expressed candidates displayed a high range of functional characteristics, illustrating the complexity and the variability of neuroplastic changes induced in the brain by psychostimulant treatment. In particular, CD81, a tetraspanin, was found to be overexpressed in the nucleus accumbens upon cocaine treatment. In situ hybridization studies allowed for localization of CD81 mRNA in the brain regions functionally related to the regulation of cardiovascular function and fluid homeostasis, most of them also expressing the neuropeptide galanin. In addition, CD81-deficient mice exhibited sex-dependent altered sensitivity to cocaine, as well as modified dopamine metabolism. One protein related to neurodegenerative processes in the brain, g-synuclein, was found to be overexpressed in the tegmentum upon cocaine treatment, suggesting a link between the neuroplasticity changes related to neural degeneration and drug addiction. Finally, one new vertebrate member of the semaphorin family and five novel rat orthologues of class 3, 4 and 7 semaphorins have been identified, whereas the application of newly developed method of Enriched Differential Display allowed for finding of semaphorin differential regulation in various brain regions upon cocaine treatment

Evaluation of a biofeedback-supported cognitive-behavioral intervention in chronic tinnitus

Den Ausgangspunkt der vorliegenden Arbeit bilden zum einen Befunde zur hohen Patientenakzeptanz des Biofeedbacks und zum anderen Nachweise der Migräneforschung hinsichtlich einer Überlegenheit der Biofeedbackmethode gegenüber kognitiv-behavioralen Ansätzen. Den theoretischen Hintergrund des zweiten Aspektes stellt die Annahme dar, dass bei Verfolgen eines somatischen Krankheitsmodells der Tinnitusentstehung vorausgehendes Biofeedback hilft, kognitive und emotionale Einflüsse auf Körperreaktionen zu erkennen. Dies begünstigt eine psychosomatische Sichtweise, die als geeigneten Ausgangspunkt für nachfolgende kognitive Methoden gilt. Mangels konkreter Nachweise der vielfach angenommenen Vorteile des Bio-feedbacks, wurde in der vorliegenden Arbeit erstmalig der differentielle Anteil dieses psychophysiolgischen Ansatzes am erwarteten Interventionseffekt unter-sucht. In der Tinnitusforschung findet sich bislang noch keine empirische Evi-denz für die relative Wirksamkeit des biofeedbackgestützten Relaxationstrain-ings im Vergleich zu einem rein kognitiv-behavioralen Vorgehen. Bei Annahme pathologischer Muskelverspannungen als Tinnitusverstärker wird von einer Beeinträchtigungsreduktion durch eine verbesserte Relaxationsfähig-keit als Copingstrategie ausgegangen. Hierfür wurden in einer kontrollierten und randomisierten Cross-over-Studie eine kognitiv-behaviorale Gruppeninterven-tion und ein Biofeedbacktraining im Einzelsetting kombiniert und gegenüberge-stellt. Forschungsberichten zufolge zeigen nämlich kognitiv-verhaltenstherapeu-tische Konzepte mit integriertem Relaxationstraining bei chronischem Tinnitus die höchsten Erfolgsraten. Die Wirksamkeitsprüfung der 12-wöchigen Interven-tion erfolgte im ambulanten Setting an 112 Tinnitusbetroffenen, die als leicht-gradig chronisch beeinträchtigt gelten. Die Datenlage zeigt Verbesserungen hinsichtlich tinnitusspezifischer Symptome (Tinnitusfragebogen) und eingesetzter Bewältigungsstrategien. Bezüglich unter-suchter physiologischer Parameter ließen sich für den Bereich M. frontalis signi-fikante Spannungsreduktionen (EMG) von Sitzungsanfang bis -ende sowie von der ersten bis zur sechsten Biofeedbackeinheit nachweisen. Die Reduktion der Muskelanspannung für den EMG-frontalis in der ersten Sitzung erwies sich als prädiktionsstark für die Reduktion der Tinnitusbelastung. Nachgewiesen sind mit den Daten eine tendenzielle Überlegenheit der psychophysiologischen ge-genüber der kognitiv-verhaltensorientierten Methode sowie eine hohe Patienten-akzeptanz der biofeedbackgestützten Relaxationsintervention.Evaluation of a biofeedback-supported cognitive-behavioral intervention in chronic tinnitus The beginning of this present paper consists of two parts: On the one hand there are findings about a high patient acceptance regarding biofeedback and on the other hand there is evidence delivered by migraine research stating that the biofeedback-method is superior to cognitive-behavioral approaches. The theoretical background of the second aspect assumes that prior biofeedback helps in recognizing cognitive and emotional influences on body reactions when following the somatic disease model of tinnitus occurrence. This approach favors a psychosomatic view that is considered to be suitable for the following cognitive methods. Due to a lack of concrete evidence regarding the assumed advantages of biofeedback, we explored the differential share of this psychophysiological approach in the expected intervention effect in this paper for the very first time. Up to date, tinnitus research is not able to present empiric evidence for the relative effectiveness of biofeedback-supported relaxation training compared to a purely cognitive-behavioral procedure. When assuming that pathological muscle tension intensifies the occurrence of tinnitus, then a coping strategy - improving the patient’s relaxation ability –would reduce his/her impairment. A cognitive-behavioral group intervention and biofeedback training in single setting were combined in a controlled and randomized cross-over study and then compared. Research reports show that cognitive behavioral therapeutic concepts with integrated relaxation training have the highest success rates in cases of chronic tinnitus. The effectiveness test of the 12-week long intervention took place in an outpatient setting and included 112 patients suffering from a light-chronic degree of tinnitus. The following is a summary of the findings: (1) The central statement of the study was that participating in the 12-week long biofeedback-supported tinnitus coping training resulted in a treatment-specific reduction of the impairment caused by tinnitus, which remained constant over time. For both defined treatment elements, cognitive-behavioral unity in the group setting and biofeedback in the individual setting, evidence for efficacy was provided. (2) While there are positive changes in the ways of coping and handling with the impairment, no improvement regarding mood, sensibility towards noise and catastrophizing was detectable. (3) The increased relaxation ability during the course of the biofeedback-relaxation training is detectable in the physiological relaxation indicator EMG and can also be detected through electrodermal activity. In the area of the m. frontalis and m. masseter, there are significant tension reductions (EMG) from the beginning of the session to the end of the session as well as from the first to the sixth biofeedback unit. (4) In regards to the differential share of the biofeedback, there is empirical evidence for psychophysiological biofeedback training having the tendency of being superior to cognitive behavioral-oriented group programs. (5) For predicting the success of treatment, the degree of severity of tinnitus as well as the tension reduction in the area of the m. frontalis in the first biofeedback session with inclusion of the partial sample RF 2 (beginning with biofeedback) seem to be strong predictors. (6) All in all, biofeedback with its supporting function in cases of documented high patient acceptance presents itself as a very suitable treatment component when being part of an outpatient behavioral-therapeutic approach in chronic tinnitus. (7) A first proposal for future studies refers to the test design used in this case. A methodic alternative, which would help in avoiding a method deficit caused by cross-over design, would be the additional definition of two parallel intervention conditions with pure behavioral therapy as well as pure biofeedback training

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Vorlageform des Erscheinungsvermerks: Anno. M.D.xxiij.||[Erfurt: Wolfgang Stürmer