Scotin, a novel p53-inducible proapoptotic protein located in the ER and the nuclear membrane

p53 is a transcription factor that induces growth arrest or apoptosis in response to cellular stress. To identify new p53-inducible proapoptotic genes, we compared, by differential display, the expression of genes in spleen or thymus of normal and p53 nullizygote mice after γ-irradiation of whole animals. We report the identification and characterization of human and mouse Scotin homologues, a novel gene directly transactivated by p53. The Scotin protein is localized to the ER and the nuclear membrane. Scotin can induce apoptosis in a caspase-dependent manner. Inhibition of endogenous Scotin expression increases resistance to p53-dependent apoptosis induced by DNA damage, suggesting that Scotin plays a role in p53-dependent apoptosis. The discovery of Scotin brings to light a role of the ER in p53-dependent apoptosis

Nanofibrous poly(lactide-co-glycolide) membranes loaded with diamond nanoparticles as promising substrates for bone tissue engineering

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Toxic Diatom Aldehydes Affect Defence Gene Networks in Sea Urchins.

Marine organisms possess a series of cellular strategies to counteract the negative effects of toxic compounds, including the massive reorganization of gene expression networks. Here we report the modulated dose-dependent response of activated genes by diatom polyunsaturated aldehydes (PUAs) in the sea urchin Paracentrotus lividus. PUAs are secondary metabolites deriving from the oxidation of fatty acids, inducing deleterious effects on the reproduction and development of planktonic and benthic organisms that feed on these unicellular algae and with anti-cancer activity. Our previous results showed that PUAs target several genes, implicated in different functional processes in this sea urchin. Using interactomic Ingenuity Pathway Analysis we now show that the genes targeted by PUAs are correlated with four HUB genes, NF-κB, p53, δ-2-catenin and HIF1A, which have not been previously reported for P. lividus. We propose a working model describing hypothetical pathways potentially involved in toxic aldehyde stress response in sea urchins. This represents the first report on gene networks affected by PUAs, opening new perspectives in understanding the cellular mechanisms underlying the response of benthic organisms to diatom exposure

Исследование эффективности брейкеров для жидкостей гидроразрыва пласта

Исследование влияния биоразлагаемых брейкеров на реологические характеристики жидкостей гидроразрыва пласта.Investigation of the effect of biodegradable breakers on the rheological characteristics of fracturing fluids

Induction of PPM1D following DNA-damaging treatments through a conserved p53 response element coincides with a shift in the use of transcription initiation sites

PPM1D (Wip1), a type PP2C phosphatase, is expressed at low levels in most normal tissues but is overexpressed in several types of cancers. In cells containing wild-type p53, the levels of PPM1D mRNA and protein increase following exposure to genotoxic stress, but the mechanism of regulation by p53 was unknown. PPM1D also has been identified as a CREB-regulated gene due to the presence of a cyclic AMP response element (CRE) in the promoter. Transient transfection and chromatin immunoprecipitation experiments in HCT116 cells were used to characterize a conserved p53 response element located in the 5′ untranslated region (UTR) of the PPM1D gene that is required for the p53-dependent induction of transcription from the human PPM1D promoter. CREB binding to the CRE contributes to the regulation of basal expression of PPM1D and directs transcription initiation at upstream sites. Following exposure to ultraviolet (UV) or ionizing radiation, the abundance of transcripts with short 5′ UTRs increased in cells containing wild-type p53, indicating increased utilization of downstream transcription initiation sites. In cells containing wild-type p53, exposure to UV resulted in increased PPM1D protein levels even when PPM1D mRNA levels remained constant, indicating post-transcriptional regulation of PPM1D protein levels

Antigenspezifische DNA-Immunisierung in Kombination mit IDO-kodierenden Plasmiden zur Therapie der experimentellen Typ I-Allergie

Derzeit stellt die allergenspezifische Immuntherapie die einzige nicht allein antisymptomatische Behandlungsform zur langfristigen Therapie von Typ I-Allergien dar, welche grundlegende Änderungen im immunologischen Geschehen induziert. Sie ist jedoch verbesserungswürdig in Bezug auf Behandlungsdauer, Erfolgschancen und Nebenwirkungen. Daher wurde in dieser Arbeit eine Strategie zur Therapie von Typ I-Allergien entwickelt und evaluiert, welche auf der Inhibition allergenspezifischer T-Zellen durch Dendritische Zellen (DC), die selektiv nach DNA-Immunisierung sowohl das relevante Allergen als auch Indolamin 2,3-dioxygenase (IDO) konstitutiv produzieren, basiert. IDO ist ein Enzym aus dem Tryptophan-Stoffwechsel, dessen Produktion durch DC einen lokalen immunsuppressiven Mechanismus induziert und in verschiedenen Situationen mit der Induktion peripherer Toleranz assoziiert ist. Zunächst wurden Plasmide hergestellt, die entweder IDO alleine oder IDO zusammen mit dem Antigen unter der Kontrolle des ubiquitär aktiven CMV- bzw. des DC-spezifischen Fascin-Promotors kodieren. Die Überprüfung der IDO-Expression durch die monocistronischen Plasmide anhand von Transfektionsexperimenten in vitro ergab, dass die IDO-Expression unter der Kontrolle des CMV-Promotors sehr viel stärker ausfiel als unter der Kontrolle des Fascin-Promotors. Nach Transfektion mit den bicistronischen Vektoren, in denen die Transgene für das Antigen und IDO durch eine IRES-Sequenz verbunden waren, war die IDO-Expression jedoch insgesamt sehr schwach. Im Rahmen der Überprüfung der Funktionalität der IDO-Expressionplasmide in vivo unter Verwendung der Genpistole wurden daher lediglich Plasmide getestet, die alleine IDO unter der Kontrolle des CMV-Promotors bzw. des Fascin-Promotors kodieren. Auch in vivo wurde eine stärkere IDO-Expression nach biolistischer Transfektion mit solchen Vektoren beobachtet, in denen der CMV-Promotor zur Expressionskontrolle verwendet wurde. Die Analyse des Einflusses einer Koexpression von IDO auf die durch biolistische Immunisierung mit einem antigenkodierenden Vektor induzierte systemische Immunantwort offenbarte einen inhibitorischer Effekt für den Fall, dass die Antigenproduktion mittels des Fascin-Promotors auf DC fokussiert war und die Expression des koapplizierten IDO-Transgens unter der Kontrolle des CMV-Promotors stand. In diesem Fall wurde eine Reduktion der antigenspezifischen IgG1- und IgG2a-Produktion, eine verringerte Sekretion von IFN-y durch restimulierte Milz- und Lymphknotenzellen sowie eine Reduktion der Zahl antigenspezifischer CD8+ Effektor-T-Zellen nachgewiesen. Im Mausmodell der IgE-vermittelten Typ I-Allergie wurde weiterhin gezeigt, dass nach prophylaktischer biolistischer Vakzinierung unter Verwendung dieser Vektorkombination eine Inhibition der durch die Vakzinierung bedingten antigenspezifischen Th1-Immunantwort ausgelöst wurde. Die Suppression der Th2-Antwort, welche durch Transfektion mit dem Antigenkodierenden Vektor unter Kontrolle des Fascin-Promotors bewirkt wurde, wurde durch Kotransfektion mit den IDO-kodierenden Vektoren aufrecht erhalten.Currently the allergen specific immunotherapy exclusively represents the not only anti-symptomatic therapeutic method for a long-term therapy of type I allergies which induces basic changes in the immunological occurrence. Nevertheless this approach is improvable regarding duration of the therapy, chances of success and adverse reactions. Therefore in this study we developed and evaluated a new strategy for a therapy of type I allergies which is based on the inhibition of allergen specific T-cells by dendritic cells (DC), which selectively and constitutively produce the relevant antigen as well as Indolamine 2,3-dioxygenase (IDO) after DNA-vaccination. IDO is an enzyme of the tryptophan catabolism. Its production by DC induces local immunosuppressive mechanisms and is associated with the induction of peripheral tolerance in various situations. Initially plasmids were created encoding either for IDO alone or for IDO and the antigen under the control of the ubiquitous active CMV- or the DC-specific fascin-promoter. Testing the IDO-expression of the monocistronic plasmids in vitro by transfection experiments showed that IDO-expression under control of the CMV-promoter was much higher than under the control of the fascin-promoter. However after transfection using the bicistronic vectors, in which the transgenes for IDO and the antigen were connected by an IRES-sequence, showed very low IDO-expression generally. So for testing the functionality of the IDO-plasmids in vivo by using the gene gun only plasmids were used encoding for IDO alone under the control of the CMV- respectively the fascin-promoter. Even in vivo the IDO-expression was higher after biolistic transfection using vectors encoding the antigen under the control of the CMV-promoter. Analyzing the influence of the IDO-coexpression to the systemic immune response after biolistic vaccination with an antigen encoding vector revealed an inhibitory effect in case that the antigen production was focused on DC by using the fascin-promoter and the expression of the co-applicated IDO-transgene was controlled by the CMV-promoter. In this case reduction of allergen specific IgG1- and IgG2a-production, reduced secretion of IFN-y by restimulated spleen- and lymph node cells as well as reduction of the amount of antigen specific CD8+effector-T-cells was determined. Furthermore in a murine model of IgE-mediated type I-allergy it was shown that after prophylactic biolistic vaccination using this vector combination the antigen specific Th1-immune response induced by the vaccination was inhibited. Suppression of the Th2-response which was achieved by the transfection with the antigen encoding vector under the control of the fascin-promoter, was sustainable after co-transfection with the IDO-encoding vector

Flexible, fast and automatic groove pass design for energy optimization using PyRolL for variable round profiles

<p>The archive contains asupplementary material to the publication: Flexible, fast and automatic groove pass design for energy optimization using PyRolL for variable round profiles; C. Renzing, M. Weiner, M. Schmidtchen and U. Prahl; Der Kalibreur; 2023. </p> <p>The content of the archive is a Python pipenv enviroment as well as a computational notebook which contains the described routine.<br>Further the corresponding presentation, which was held at the conference can be found.</p&gt