Expression regulation of MAO isoforms in monocytic cells in response to Th2 cytokines

Background: Th2-cytokines, such as interleukins-4 and –13 (IL-4, IL-13), have been identified as alternative stimuli of monocytes/macrophages. We have recently profiled the gene-expression pattern of IL-4-teated human peripheral monocytes and found that 15-lipoxygenase-1 (15-LOX1) and monoamine oxidase A (MAO-A) are among the five most strongly upregulated gene products in IL-4-treated cells. Transfection of monocytic cells (U937) with 15-LOX1 also induced MAO-A expression. These data suggested that 15-LOX1 products might play a role in the IL4-induced signaling cascade leading to expression of MAO-A in human monocytes. Material/Methods: To test this hypothesis we incubated wild-type and 15-LOX1-transfected U937 cells with different concentrations of either IL-4 or 15-LOX-products [13S-H(p)ODE, 15S-H(p)ETE] and quantified the expression of 15-LOX1, MAO-A, and MAO-B by activity assays and real-time RT-PCR. Results: Wild-type U937 cells express neither MAO-A nor MAO-B, but after three days of IL4 treatment, MAO-A mRNA was detected. A similar isoform-specific expression of MAO-A mRNA was observed when U937 cells were transfected with 15-LOX1 or when the cells were incubated with primary 15-LOX1 products (hydroperoxy fatty acids) or H2O2. In contrast, the corresponding hydroxy fatty acids were ineffective. Conclusions: These data indicate that increased intracellular peroxide concentrations (oxidative stress) induce MAO-A expression in monocytes/macrophages, which normally do not express the enzyme. Our findings also suggest that IL-4-induced upregulation of MAO-A expression in human peripheral monocytes may proceed via 15-LOX1-dependent and 15-LOX1-independent pathways. The biological role of MAO-A expression for monocyte function is discussed

Lippen-Kiefer-Gaumen-Spalten: Individuelle Analyse der Lippenspalte durch 3-D-Lasertopometrie

Zusammenfassung: Hintergrund: Mit Gipsmodellen und Fotografien ist die dreidimensionale Analyse einer Lippen-Kiefer-Gaumen-Spalte meist nur unzureichend möglich. Ziel der vorliegenden Studie war es daher, die 3-D-Lasertopometrie auf ihre Anwendbarkeit zur dreidimensionalen Weichgewebserfassung bei Patienten mit Lippen-Kiefer-Gaumen-Spalten zu testen. Patienten und Methode: Bei 20 Patienten (3-35 Jahre), die eine einseitige, nicht operierte Lippen-, Lippen-Kiefer- oder Lippen-Kiefer-Gaumen-Spalte aufwiesen, wurde mit einem 3-D-Laserscanner die Gesichtsoberfläche prä- und postoperativ dreidimensional erfasst. Die dabei erzeugten digitalen Datensätze wurden in einer virtuellen Umgebung metrisch analysiert und anhand von Quotienten größenunabhängig wiedergegeben. Sie dienten der Auswahl der Operationstechnik und der Beurteilung des Operationsergebnisses. Ergebnisse: Mit dem 3-D-Laserscanner wurden 3-D-Oberflächen guter Qualität erstellt, die sich im Millimeterbereich ausmessen ließen. Die dreidimensionale Spaltmorphologie konnte in den Datensätzen reproduzierbar mit Landmarks versehen und vermessen werden. Auch die postoperative Symmetrie ließ sich so kontrollieren und objektivieren. Als nachteilig erwiesen sich die relativ lange Messzeit und die Notwendigkeit zur Kombination mehrerer Ansichten. Schlussfolgerung: Das vorgestellte 3-D-Laserverfahren ermöglicht eine präzise dreidimensionale Weichteilanalyse der Lippen- und Nasenregion bei Spaltpatienten. Es eignet sich jedoch nur bedingt für lebhafte Säuglinge und unkooperative Patiente

Adhesion to carbon nanotube conductive scaffolds forces action-potential appearance in immature rat spinal neurons

In the last decade, carbon nanotube growth substrates have been used to investigate neurons and neuronal networks formation in vitro when guided by artificial nano-scaled cues. Besides, nanotube-based interfaces are being developed, such as prosthesis for monitoring brain activity. We recently described how carbon nanotube substrates alter the electrophysiological and synaptic responses of hippocampal neurons in culture. This observation highlighted the exceptional ability of this material in interfering with nerve tissue growth. Here we test the hypothesis that carbon nanotube scaffolds promote the development of immature neurons isolated from the neonatal rat spinal cord, and maintained in vitro. To address this issue we performed electrophysiological studies associated to gene expression analysis. Our results indicate that spinal neurons plated on electro-conductive carbon nanotubes show a facilitated development. Spinal neurons anticipate the expression of functional markers of maturation, such as the generation of voltage dependent currents or action potentials. These changes are accompanied by a selective modulation of gene expression, involving neuronal and non-neuronal components. Our microarray experiments suggest that carbon nanotube platforms trigger reparative activities involving microglia, in the absence of reactive gliosis. Hence, future tissue scaffolds blended with conductive nanotubes may be exploited to promote cell differentiation and reparative pathways in neural regeneration strategies

Altered Activation of Innate Immunity Associates with White Matter Volume and Diffusion in First-Episode Psychosis

First-episode psychosis (FEP) is associated with inflammatory and brain structural changes, but few studies have investigated whether systemic inflammation associates with brain structural changes in FEP. Thirty-seven FEP patients (median 27 days on antipsychotic medication), and 19 matched controls were recruited. Serum levels of 38 chemokines and cytokines, and cardiovascular risk markers were measured at baseline and 2 months later. We collected T1-and diffusion-weighted MRIs with a 3 T scanner from the patients at baseline. We analyzed the association of psychosis-related inflammatory markers with gray and white matter (WM) volume using voxel-based morphometry and WM diffusion using tract-based spatial statistics with whole-brain and region-of-interest (ROI) analyses. FEP patients had higher CCL22 and lower TGFa, CXCL1, CCL7, IFN-alpha 2 and ApoA-I than controls. CCL22 decreased significantly between baseline and 2 months in patients but was still higher than in controls. The association between inflammatory markers and FEP remained significant after adjusting for age, sex, smoking and BMI. We did not observe a correlation of inflammatory markers with any symptoms or duration of antipsychotic treatment. Baseline CCL22 levels correlated negatively with WM volume and positively with mean diffusivity and radial diffusivity bilaterally in the frontal lobes in ROI analyses. Decreased serum lan association between circulating chemokine levels and WM in FEP patients. Interestingly, CCL22 has been previously implicated in autoimmune diseases associated with WM pathology. The results suggest that an altered activation of innate immunity may contribute to WM damage in psychotic disorders.evel of ApoA-I was associated with smaller volume of the medial temporal WM. In whole-brain analyses, CCL22 correlated positively with mean diffusivity and radial diffusivity, and CXCL1 associated negatively with fractional anisotropy and positively with mean diffusivity and radial diffusivity in several brain regions. This is the first report to demonstratePeer reviewe

Differential modulation of the expression of pro- and anti-inflammatory gene products

0\. Titelblatt, Danksagung und Verzeichnisse DANKSAGUNG I. INHALTSVERZEICHNIS II. ABBILDUNGSVERZEICHNIS III. TABELLENVERZEICHNIS IV. ABKÜRZUNGSVERZEICHNIS 1\. EINLEITUNG 1.1. Die Entzündung ist Teil der Immunantwort 1.2. Eikosanoide sind effektive Entzündungsmediatoren 1.3. Lipoxygenasen sind lipidperoxidierende Enzyme 1.4. Monoaminoxidasen sind amin-oxidierende Enzyme, die im Stoffwechsel von Neurotransmittern bedeutsam sind 1.5. Das humane Immundefizienz-Virus (HIV) 1.6. Zielstellung 2\. MATERIAL UND METHODEN 2.1. Material 2.2. Methoden 3\. ERGEBNISSE 3.1. Veränderungen im mRNA-Muster humaner Monozyten durch die Th2-Zytokine Interleukin-4/13 3.2. Veränderungen im Protein-Expressionsmuster humaner Monozyten durch die Th2-Zytokine Interleukin-4/13 3.3. IL-4-induzierte Hochregulation der Expression anti-inflammatorischer Gene 3.4. IL-4-induzierte Herunterregulation der Expression pro-inflammatorischer Gene 3.5. Parallele Induktion von 15-LOX-1 und MAO-A nach Th2-Zytokinstimulierung 3.6. Einfluss der 15 LOX-Überexpression auf die Expression von HIV- Korezeptoren 4\. DISKUSSION 4.1. Methoden der Transkriptomanalyse 4.2. Die Rolle von Monozyten/Makrophagen bei der Entzündungsreaktion 4.3. Kurzcharakterisierung der am stärksten hochregulierten Genprodukte 4.4. Kurzcharakterisierung der am stärksten herunterregulierten Genprodukte 4.5. Kurzcharakterisierung einiger anti-inflammatorischer Gene 4.6. Kurzcharakterisierung klassischer pro-inflammatorischer Gene 4.7. Regulation der Genexpression durch Veränderungen des zellulären Redox- Status 5\. ZUSAMMENFASSUNG 6\. SUMMARY 7\. LITERATURVERZEICHNIS 8\. PUBLIKATIONS-LISTEMonozyten spielen eine duale Rolle bei der Pathogenese der Entzündungsreaktion. In der akuten Entzündungsphase vermitteln sie überwiegend pro-inflammatorische Wirkungen, was u.a. durch das Muster der monozytärer Botenstoffe (Zytokine, Eikosanoide) sichtbar wird. Während in der Heilungsphase (inflammatory resolution) verändert sich jedoch der Phänotyp der im Entzündungsareal vorkommenden Monozyten dahingehend, dass sie nun vorwiegend anti-inflammatorische Mediatoren synthetisieren. Damit wird die akute Entzündungsreaktion aktiv beendet und die ursprüngliche Struktur des entzündlich veränderten Gewebes wiederhergestellt. Um zu untersuchen, ob Th2-Zytokine (IL-4 und IL-13) einen solchen Phänotypwechsel von Monozyten induzieren können, wurden periphere Blutmonozyten in Gegenwart von IL-4 oder IL-13 für 72 Stunden in vitro kultiviert und anschließend zelluläre Expressionsprofile (mehr als 16.000 Genen) mittels Mikroarray-Analyse erstellt. Die acht am stärksten hochregulierten (bis zu 290-fache Hochregulation) Genprodukte waren: 15-LOX-1, FN1, CD23, CD1C, MAO-A, F13A1, MS4A4A und CCL22. Im Gegensatz dazu wurden IFI44L, SN, CLGN, ORM1, C1S, INDO, ISG20 und KCNJ15 am stärksten herunterreguliert (bis zu 100-fache Herunterregulation). Weiterhin wurde unter unseren experimentellen Bedingungen die monozytäre Synthese klassischer pro-inflammatorischer Zytokine (IL-1, IL-6, IL-8, IL-18, CCL2, TNFα) herunterreguliert, und es kam zu einer Verminderung der Expression von Enzymen (5-LOX, LTC4-Synthase, COX-2), die an der Synthese pro-inflammatorischer Eikosanoide beteiligt sind, und von Korezeptoren (CD4, CXCR4, CCR5) des humanen Immundefizienz-Virus-1 (HIV-1). Die Expression von Proteinen, denen man anti-inflammatorische Eigenschaften zuschreibt (TIMP, ANXA1, IL-10, HMOX1) wurden dagegen hochreguliert. Neben der Regulation der 15-LOX-1, die unter den Genprodukten in Monozyten nach IL-4-Stimulierung das am stärksten hoch regulierte war, war die 54-fache Erhöhung der MAO-A Expression besonders bemerkenswert, da es für eine monozytäre Expression dieses Enzyms bislang keinen experimentellen Hinweis gab. Unsere Daten belegen jedoch, dass die MAO-A in IL4-behandelten Monozyten auf hohem Niveau exprimiert wird und dass das Enzym eine Rolle bei der Entzündungsheilung spielen könnte, da sie für den Abbau einiger pro- inflammatorischer biogener Amine verantwortlich ist. Weiterführende Experimente zum Mechanismus der Expressionsregulation der MAO-A und der HIV- Korezeptoren deuteten auf eine wichtige Rolle des zellulären Hydroperoxidtonus (zelluläre Konzentration von Hydroperoxiden) hin. So konnte die IL-4-induzierte Hochregulation der MAO-A Expression und die IL-4-abhängige Senkung der zellulären Konzentration von HIV-1-Korezeptoren auch durch eine Erhöhung des zellulären Hydroperoxidtonus ausgelöst werden. Damit könnte es sich zumindest bei einem Teil der IL-4-regulierbaren Gene um redox-abhängige Expressionsregulationen handeln. Da die enzymatische Aktivität der 15-LOX-1 zu einer Erhöhung des zellulären Oxidationsstatus führen kann, scheint die IL-4-induzierte Hochregulation der Expression dieses Lipid-peroxidierenden Enzyms über die IL-4-induzierte Regulationskaskade besonders bedeutsam zu sein.Monocytes play a pivotal role in the pathogenesis of the inflammation. In the acute phase of inflammation they obtain predominantly pro-inflammatory effects, which become visible in the expression of second messengers (cytokines, eicosanoids). During the inflammatory resolution phase the phenotype of the monocytes occurring in the inflammation area changes in such a way that they now synthesize predominantly anti-inflammatory mediators. Thus the acute inflammation reaction is terminated actively and the original structure of the inflammatory changed fabric is restored. In order to examine, whether Th2-cytokines (IL-4 and IL-13) can induce such a phenotype-switch, peripheral monocytes were cultivated in presence of IL-4 or IL-13 for 72 hours in vitro afterwards cellular expression profiles (more than 16,000 genes) were obtained by means of microarray analysis. The eight most strongly up-regulated (up to 290-fold up-regulation) genes were: 15-LOX-1, FN1, CD23, CD1C, MAO A, F13A1, MS4A4A and CCL22. In contrast, IFI44L, SN, CLGN, ORM1, C1S, INDO, ISG20 and KCNJ15 were the most strongly down-regulated (up to 100-fold down- regulation) genes. Under our experimental conditions classical pro- inflammatory cytokines (IL-1, IL-6, IL-8, IL-18, CCL2, TNFα) become down- regulated. And in addition we observed a reduction of the expression of enzymes (5-LOX, LTC4-Synthase, COX-2), which are involved in the synthesis of pro-inflammatory eicosanoids and of co-receptors (CD4, CXCR4, CCR5) for the human immunodeficiency virus-1 (HIV-1). In contrast, the expression of proteins with anti-inflammatory characteristics (TIMP, ANXA1, IL-10, HMOX1) are up-regulated. In addition to the up-regulation of the 15-LOX-1, which was the most strongly regulated gene in monocytes after IL-4 stimulation, we observed a 54-fold increase of MAO-A expression. This was particularly remarcable, as there was no experimental reference for a monocyte expression of this enzyme so far. However our data prove that the MAO-A in IL4-treated monocytes is expressed ad high level and that the enzyme could play a crucial role during the inflammatory resolution phase, as it is responsible for the degradation of some pro-inflammatory biogenic amines. Further experiments on the mechanisms of the expression regulation of the MAO-A and the HIV co- receptors indicated an important role of the cellular hydroperoxide tonus. So the IL-4-induced up-regulation of the MAO-A expression and the IL-4-dependent decrease of the cellular concentration of HIV-1 co-receptors could be induced by an increase of the cellular hydroperoxide tonus. This suggest, that at least a part of the IL-4-regulated genes is redox-dependent regulated. Since the enzymatic activity of the 15-LOX-1 can lead to an increase of the cellular oxidation status, the IL 4-induced up-regulation of the expression of this lipid-peroxiding enzyme by means of the IL-4-induced regulation cascade seems to be particularly important

Potential Effects on Human Safety and Health from Infrasound and Audible Frequencies Generated by Vibrations of Diesel Engines Using Biofuel Blends at the Workplaces of Sustainable Engineering Systems

Employees involved in various occupational environments that include vibration machines and any kind of vehicles are adversely subjected to multiple source noise. Thus, the corresponding noise frequencies (and mainly the infrasound ones) present high interest, especially from the viewpoint of sustainability, due to the potential effects on human safety and health (H_S&H) in sustainable engineering projects. Moreover, the occupational safety and health (OSH) visualization (a fact of unveiling the social dimension of sustainability) of occupational workplaces (by evaluating the infrasound and audible noise frequencies generated by diesel engines) could help a safety officer to lessen crucial risk factors in the OSH field and also to protect, more efficiently, the employees by taking the most essential safety measures. This study (i) suggests a technique to determine the infrasound and audible sound frequencies produced due to vibrations of diesel engines, by using biofuels (i.e., sustainable utilization of resources), in order to evaluate potential effects on human safety and health at the workplaces of sustainable engineering projects, and (ii) it ultimately aims to contribute to the improvement of the three “sustainability pillars” (economy, social, and environmental). Therefore, it provides experimental results of the frequency of the noise (regarding the infrasound and audible spectrum) that a diesel motor generates by vibration, in the frame of using different engine rpms (850, 1150, and 2000) and a variety of biofuel mixtures (B20-D80, B40-D60, B60-D40, and B80-D20). The article shows that the fuel blend meaningfully affects the generated noise, and more particularly, the usage of biofuel blends coming from mixing diesel oil with biodiesel (a fact of the emerging environmental dimension of sustainability) can produce various noise frequencies, which are determined in the infrasound and audible spectra (~10–23 Hz). The suggested technique, by ameliorating the OSH situation, doubtless will help enterprises to achieve the finest allocation of limited financial resources (a fact corresponding to the economic dimension of sustainability), allowing financial managers to have more available budget for implementing other risk-reduction projects

Treatment of chronic venous disorder: A comprehensive review

Chronic venous disorder (CVD) is highly prevalent vascular disorder affecting up to 45% of the general population, with clinical manifestations ranging from teleangiectasias to venous leg ulcers (VLUs). We examined the currently available data in order to provide an updated, comprehensive review on treatment options of CVD. We searched MEDLINE, Cochrane, Scopus, EMBASE, ClinicalTrials, and OpenGrey databases for relevant articles in English published until November 2020. Compression treatment is the mainstay of conservative treatment. Pharmacological treatment can provide significant symptomatic relief and hence it should be considered as part of conservative treatment. Transcutaneous Lacer treatment (TCL) is a safe and effective alternative option to sclerotherapy for treatment of C1 stage. High ligation and stripping (HL/S), ultrasound-guided foam sclerotherapy (UGFS), endovenous thermal ablation (EVTA) systems and non thermal non tumescent ablation (NTNT) systems are safe and efficacious first-line options for treatment of saphenous insufficiency. Interventional treatment of co-existing incompetent perforator veins (IPVs) is not supported by contemporary evidence. Regarding deep venous insufficiency (DVI), treatment of symptomatic femoroiliocaval occlusive venous disease refractory to conservative treatment with percutaneous transluminal venoplasty stenting has produced encouraging results

Εξατομικευμένη δημιουργία ενός platformer παιχνιδιού μέσω PCGML αλγορίθμων

Summarization: In recent years, the game development industry has shown an enormous interest in the field of Procedural Content Generation (PCG). Especially the autonomous generation of levels for video games has become a popular subject of study. Generative Adversarial Networks (GANs) are a machine learning method that has been used over the last decade and proved to be capable of generating media content and even game content. For this thesis, a new video game that combines the characteristics of a Platformer and a Dungeon Crawler has been developed in Unity from which a handcrafted dataset has been created. A GAN was successfully trained on this dataset and was therefore able to generate new level rooms. Since the core of this work was to generate rooms whose difficulty scale is adjusted based on the player’s performance, the input latent vector to the generative model was found by using an Evolutionary Strategy. Specifically, the Covariance Matrix Adaptation Evolutionary Strategy (CMA-ES) was applied which searched through the latent space of the GAN by utilizing the static characteristics of each room in form of a fitness function. The output room of this process was then given a difficulty score by evaluation and placed in the correct spot of the 3x3 grid which constitutes the whole level. The experiments of this thesis are resulted from having a variety of player models and actual players play the game and show how capable the trained GAN model is in generating novel example outputs that fit the player’s performance.Περίληψη: Τα τελευταία χρόνια, η βιομηχανία ανάπτυξης παιχνιδιών έχει δείξει τεράστιο ενδιαφέρον στον τομέα της διαδικασίας αυτόματης δημιουργίας περιεχομένου (PCG). Ειδικά η αυτόνομη παραγωγή επιπέδων για βιντεοπαιχνίδια έχει γίνει ένα δημοφιλές αντικείμενο μελέτης. Τα Γενετικά Ανταγωνιστικά Δίκτυα (GANs) είναι μια μέθοδος μηχανικής μάθησης που έχει χρησιμοποιηθεί την τελευταία δεκαετία και αποδείχθηκε ικανή να δημιουργεί περιεχόμενο πολυμέσων και περιεχόμενα παιχνιδιών. Για αυτήν τη εργασία αυτήν, ένα νέο βιντεοπαιχνίδι που συνδυάζει τα χαρακτηριστικά ενός Platformer και ενός Dungeon Crawler αναπτύχθηκε στην Unity από το οποίο δημιουργήθηκε ένα σύνολο δεδομένων. Ένα GAN εκπαιδεύτηκε με επιτυχία στο συγκεκριμένο σύνολο δεδομένων και κατά συνέπεια μπόρεσε να δημιουργήσει νέα επίπεδα του παιχνιδιού. Δεδομένου ότι ο πυρήνας αυτής της εργασίας ήταν να δημιουργηθούν επίπεδα των οποίων η κλίμακα δυσκολίας προσαρμόζεται με βάση την απόδοση του παίκτη, το διάνυσμα εισόδου στο μοντέλο δημιουργίας βρέθηκε χρησιμοποιώντας μια Εξελικτική Στρατηγική. Συγκεκριμένα, εφαρμόστηκε το Covariance Matrix Adaptation Evolutionary Strategy (CMA-ES) που αναζητούσε τον λανθάνοντα χώρο του GAN χρησιμοποιώντας τα στατικά χαρακτηριστικά κάθε επιπέδου με τη μορφή μιας συνάρτησης καταλληλότητας. Στη συνέχεια, στην έξοδο αυτής της διαδικασίας δόθηκε βαθμολογία δυσκολίας με αξιολόγηση και τοποθετήθηκε στο σωστό σημείο του πλέγματος 3x3 που αποτελεί ολόκληρο το επίπεδο παιχνιδιού. Τα πειράματα αυτής της εργασίας προέρχονται από μια ποικιλία μοντέλων παικτών και πραγματικών παικτών οι οποίοι έπαιξαν το παιχνίδι και έδειξαν πόσο ικανό είναι το εκπαιδευμένο μοντέλο GAN στη δημιουργία νέων επιπέδων που ταιριάζουν στην απόδοση του παίκτη