Abstract The principal aim of a population-based cross-sectional survey was to generate information on the lifetime occurrence of prostatitis in Finnish men and their exposure to the disease, and also on the influence of prostatitis-related fears and disturbances on their sexual life. A second aim was to develop and clinically validate a new diagnostic tool for differential diagnosis between the forms of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), especially between patients belonging to categories IIIA and IIIB in the new NIH (National Institutes of Health) clinical classification. Altogether 1832 men out of 2500 aged 20–59 years chosen randomly from the two most northerly provinces of Finland (Oulu and Lapland) participated in the epidemiological study, a response rate of 75%. The overall lifetime prevalence of prostatitis was 14.2%. The risk of having had the disease increased with age, being 1.7 times greater in the men aged 40–49 years than in those aged 20–39 years, and 3.1 times greater in those aged 50–59 years. More than a quarter of the 261 men who had or had had prostatitis symptoms (27%) suffered from them at least once a year, while 16% suffered from chronic prostatitis symptoms throughout the year. 63% of the men with prostatitis had their worst symptoms during the wintertime (November–march). 17% of the men with chronic prostatitis reported a constant fear of undetected prostate cancer. Erectile dysfunction was reported by 43% of the symptomatic men and decreased libido by 24%. Self-assessment of personality showed that the men with prostatitis were more often busy and nervous and had a meticulous attitude to life and problems than were the non-symptomatic men. 197 patients with chronic prostatitis/chronic pelvic pain syndrome participated in three clinical case-control studies during the years 1995–2000, at Oulu University Hospital, the District Hospital of Oulainen and Seinäjoki Central Hospital. The first prostatic tissue pressure measurement (PTPM) study included 34 patients and 9 controls. A novel method was developed to measure intraprostatic tissue pressure with a Stryker® intracompartmental pressure monitor. The PTPM showed a clear increase (p < 0.001) in the patients with symptoms of prostatitis and benign prostatic enlargement (BPE) relative to the controls and the patients with BPE but without pain symptoms. The second PTPM study included 42 patients with chronic prostatitis symptoms without significant BPE and 12 new controls. Significantly higher pressure readings (p < 0.001) were recorded at all three measurement points in the patients than in the controls. 48 new patients and 12 new controls were enrolled for the third PTPM study, the purpose of which was to confirm the results of the previous ones and to compare the prostatic tissue pressures of two clinical groups (IIIA and IIIB). The prostatic tissue pressure was again significantly higher in the patients with chronic prostatitis symptoms than in the controls (p < 0.001). An interesting finding was that prostatitis patients belonging to clinical category IIIA had significantly higher tissue pressures (p < 0.01) than those in category IIIB, probably reflecting more severe inflammation in the prostatic tissue. This new PTPM method provides a more precise and/or exact tool for differential diagnosis between the forms of pelvic pain and CP/CPPS.

The current industry landscape is witnessing a trend towards high-mix production, which requires a reconsideration of the existing production systems. Although high levels of automation have been achieved in the industry, the traditional automated production line, designed for mass production of homogeneous goods, is not well-suited for high-mix production. To address this situation, flexible and adaptable alternatives have been sought to replace the inflexible and rigid traditional production lines. One of the proposed solutions is the combination of digital technology and physical automation, creating a highly connected and intelligent production environment. Such an environment requires the implementation of a cyber-physical production system that integrates Industry 4.0 technologies, such as Automated Guided Vehicles (AGVs) and flexible robots. This system enables a set of robots to perform different tasks instead of being exclusively dedicated to a specific task, making it moreadaptable and flexible. The integration of advanced technologies, such as AGVs and Cyber-Physical Production Systems (CPPS), can significantly enhance workflow optimization, reduce production times, and enable flexible layout adaptations to cater to the specific requirements of different products in the production line. Furthermore, it can facilitate better control of information and enable real-time monitoring of the production process, leading to improved production efficiency and quality. To demonstrate the potential of such a system, a virtual commissioning of a fully innovative production line has been carried out, encompassing all the previously mentioned technologies and elements. The virtual commissioning of the production line serves as a proof-of-concept for the cyber-physical production system and its ability to provide a highly connected, intelligent, and adaptable production environment. / <p>Utbytesstudenter</p>

Orientador: Marlus Chorilli / Resumo: O diabetes mellitus (DM) é uma síndrome metabólica caracterizada por deficiência na produção/secreção pancreática de insulina e/ou resistência à ação do hormônio nos tecidos alvo, resultando em hiperglicemia. Diversas pesquisas têm desencadeado o desenvolvimento de novos sistemas de administração de insulina que possibilitem a utilização de vias alternativas à parenteral, com destaque à administração de insulina por via nasal. Esta via tem-se mostrado promissora, pois pode promover uma rápida absorção do fármaco e aumentar a sua biodisponibilidade. Entretanto, existem mecanismos de depuração mucociliar que limitam a administração de fármacos, além da baixa permeabilidade do epitélio nasal, o qual dificulta a absorção de fármacos com alto massa molar. Uma estratégia para vencer tais barreiras é a utilização de sistemas nanoestruturados (lipossomas), pois são amplamente utilizados para o aperfeiçoamento da potencialização da ação terapêutica de fármacos. Além disso estes lipossomas foram funcionalizados com peptídeos de transdução de membrana (CPPs), tais como os peptídeos TAT e Penetratin (PNT), que atuam como promotores da penetração e absorção do fármaco, com posterior dispersão em hidrogel de hidroxietilcelulose. O objetivo deste trabalho foi desenvolver e caracterizar lipossomas contendo solução de insulina, funcionalizados com CPPs (TAT e PNT) e dispersos em hidrogel, avaliar o potencial pela via nasal, in vivo, para a melhora dos níveis séricos e efeito hipoglicemiante d... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

Les peptides vecteurs ou CPP sont de petits peptides, en général de taille inférieure à 30 acides aminés. Parmi les nombreux CPP décrits dans la littérature, les peptides riches en arginine ont fait l'objet d'une attention particulière. Plusieurs modifications chimiques du squelette peptidique conduisant à une distribution spatiale différente des groupes fonctionnels, ou encore l'introduction de chaînes aliphatiques ont été effectuées pour accroitre la capacité du peptide à traverser la membrane de la cellule. L'objectif de ce travail a été le développement de nouveaux peptides vecteurs cycliques contenant un domaine cationique minimal et pouvant être acylés par une chaîne aliphatique. Quinze nouveaux transporteurs cycliques, dont les peptides vecteurs classiques Pénétratine et R6W3ont été synthétisés. La cyclisation tête-queue par ligation chimique native a été rendue possible par l'introduction d'un résidu cystéine et d'une fonction thioester (ou précurseur) respectivement aux extrémités N et C-terminales des différentes séquences de CPP. Leur aptitude à transporter le peptide bioactif PKCi dans des cellules CHO a été évaluée par quantification de la cargaison internalisée en utilisant la spectrométrie de masse MALDI-TOF. Les résultats indiquent une meilleure internalisation essentiellement par voie d'endocytose dépendante des glycosaminoglycanes, suite à la cyclisation des CPP comparés à leur version linéaire. De toute la série des lipopeptides testés dans ce projet, deux séquences se distinguent par leur capacité remarquable à franchir les membranes cellulaire : les CPP [C12-R4] et [C12-R7]. / Cell-penetrating peptides (CPPs) are short, cationic or amphipatic peptides, usually containing less than 30 amino acids, which are able to deliver various bioactive cargoes inside cells. Among the many CPPs described in the literature, the arginine-rich peptides have attracted particular attention. Several chemical modifications of the peptide backbone leading to different spatial distributions of the CPP functional groups, or the introduction of aliphatic chains have been made to enhance their internalization efficiency. The aim of this work was the synthesis of new cyclic CPPs containing a minimal cationic domain and their functionalisation with an aliphatic chain. We have synthesised a small library of fifteen new cyclic carriers including the classical CPPs Penetratin and R6W3 using native chemical ligation (NCL) in solution. The introduction of an N-terminal Cys residue and of a C-terminal thioester (or precursor) in the initial linear peptide sequence allowed the head-to-tail cyclisation. The efficiency of cargo delivery in CHO cells was measured by MALDI-TOF mass spectrometry. We found that cyclisation of CPPs improved their internalisation efficiency mostly by glycosaminoglycan-dependent endocytosis. Among the whole series of lipopeptides tested in this project, two sequences are distinguished by their remarkable ability to cross cellular membranes: the peptides [C12-R4] and [C12-R7].

Chronic pelvic pain is a condition which receives less attention in men than in women. It is often difficult to diagnose and more difficult to treat. The new classification of prostatitis and its variants has introduced the term ‘chronic pelvic pain syndrome’ which underlines the difficulties in dealing with this disorder which may represent a variety of chronically painful conditions with a large functional component. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Monocytes and macrophages are mediator cells of cartilage and bone erosion in the synovia of rheumatoid arthritis (RA) patients due to secretion of the inflammatory cytokine Interleukin-1β (IL-1β). Calcium, phosphate and fetuin are liberated from the affected bone matrix, and the formation of calciproteinparticles (CPPs) is likely. IL-1β production in monocytes in vitro is stimulated by high concentrations of extracellular calcium. Additionally, the rise of extracellular calcium concentrations leads to increased macropinocytosis in mononuclear phagocytes. Flow cytometry analyses in this study show that peripheral blood monocytes from patients with RA perform more calcium stimulated macropinocytosis of the fluorescent dye calcein than monocytes from healthy donors. Stimulation of monocytes with calcium and preformed CPPs leads to more IL-1β production, quantified using ELISA, by monocytes from RA patients. Experiments with macrophages show similar results. Furthermore calcium-stimulated macropinocytosis and IL-1β secretion are significantly positively correlated. However, there was no connection of in vitro findings and the severity of RA in patients.:Abbildungsverzeichnis IV Tabellenverzeichnis VI Abkürzungsverzeichnis VII 1 Einleitung 1 1.1 Rheumatoide Arthritis 1 1.1.1 Epidemiologie und Klinik der rheumatoiden Arthritis 1 1.1.2 Ätiopathogenese der rheumatoiden Arthritis 1 1.2 Monozyten und Makrophagen 3 1.2.1 Inflammasomaktivierung und Interleukin-1β-Sekretion in Monozyten und Makrophagen 4 1.2.2 Makropinozytose in Monozyten und Makrophagen 6 1.2.3 Beitrag der Monozyten und Makrophagen zur rheumatoiden Arthritis 7 1.3 Kalzium – lokale Dysregulation trotz systemischer Regulation 9 1.3.1 Entstehung von Kalziumproteinpartikeln 10 1.3.2 Kurzportrait des G-Protein-gekoppelten Kalziumrezeptors CaSR 11 2 Fragestellungen 13 3 Forschungsdesign, Material und Methoden 15 3.1 Forschungsdesign 15 3.2 Materialien 15 3.2.1 Laborgeräte 16 3.2.2 Verbrauchsmaterialien 17 3.2.3 Materialien und Chemikalien 17 3.2.4 Medien, Lösungen und Puffer 19 3.2.5 Stimulanzien und Inhibitoren 20 3.2.6 Fluoreszenzfarbstoffe 20 3.2.7 Software 20 3.3 Methoden 21 3.3.1 Separation von PBMCs mittels Ficolldichtegradientenzentrifugation 21 3.3.2 Separation von Monozyten mittels negativer Magnetseparation 22 3.3.3 Differenzierung von Monozyten zu Makrophagen in Zellkulturbeuteln 23 3.3.4 Makropinozytose von Monozyten und Makrophagen in der Durchflusszytometrie 23 3.3.4.1 Makropinozytose von Calcein in Monozyten 24 3.3.4.2 Makropinozytose fluoreszenzgefärbter Kalziumproteinpartikel in Monozyten und Makrophagen 25 3.3.4.3 Inhibition der Makropinozytose in Monozyten 26 3.3.4.4 Auswertung der am Durchflusszytometer generierten Rohdaten mit FlowJo 26 3.3.5 Makropinozytose von Monozyten in der Fluoreszenzmikroskopie 28 3.3.6 Bestimmung der Interleukin-1β-Produktion von Monozyten und Makrophagen mittels ELISA 29 3.3.7 Erhebung des DAS28 33 3.3.8 Bestimmung von Laborparametern 33 3.4 Statistische Auswertung 33 4 Ergebnisse 35 4.1 Charakterisierung der Kohorten 35 4.2 Vorversuche zur Auswahl eines geeigneten Fluoreszenzfarbstoffes für die Detektion der Makropinozytose 37 4.3 Stimulation von Monozyten mit Kalzium zur Makropinozytose und Interleukin-1β-Produktion 39 4.3.1 Kalziumstimulierte Calceinaufnahme von Monozyten 39 4.3.2 Kalziumstimulierte Interleukin-1β-Produktion von Monozyten 44 4.4 Stimulation von Monozyten mit Kalzium zur Makropinozytose und Interleukin-1β-Produktion unter Zugabe von Kalziumproteinpartikeln 47 4.4.1 Kalziumstimulierte Aufnahme fluoreszierender Kalziumproteinpartikel 47 4.4.2 Kalziumstimulierte Interleukin-1β-Produktion in Monozyten unter Zugabe von Kalziumproteinpartikeln 51 4.5 Stimulation von Makrophagen mit Kalzium zur Makropinozytose und Interleukin-1β-Produktion 53 4.5.1 Kalziumstimulierte Makropinozytose von fluoreszierenden Kalziumproteinpartikeln in Makrophagen 53 4.5.2 Kalziumstimulierte Interleukin-1β-Produktion mit und ohne Zugabe von Kalziumproteinpartikeln in Makrophagen 54 4.5.3. Visualisierung von Monozyten und Makrophagen nach 16 Stunden Inkubation 57 4.6 Korrelation zwischen kalziumstimulierter Makropinozytose und Interleukin-1β-Sekretion 59 5 Diskussion 61 5.1 Kalziumstimulierte Makropinozytoseaktivität von Monozyten und Makrophagen 61 5.2 Kalziumstimulierte Interleukin-1β-Sekretion von Monozyten und Makrophagen 64 5.2.1 Auswirkung der Phosphatkonzentration im Zellkulturmedium auf die kalziumstimulierte Interleukin-1β-Sekretion von Monozyten und Makrophagen 65 5.2.2 Kalziumstimulierte Interleukin-1β-Sekretion von Monozyten und Makrophagen von RA-Patienten und Kontrollprobanden 66 5.3 Zusammenhang von kalziumstimulierter Makropinozytose und Interleukin-1β-Sekretion in Monozyten und Makrophagen von RA-Patienten und Kontrollprobanden 70 5.4 Ausblick und offene Fragen 71 6 Zusammenfassung der Arbeit 73 8 Erklärung über die eigenständige Abfassung der Arbeit 88 9 Danksagung 89

<p>Pathogenic bacteria are evolving to drug resistant strains at alarming rates. The threat posed by drug resistant bacterial infections emphasize the need to establish new antimicrobial agents. Of immediate concern regarding the dangers of antibiotic resistance is the existence of intracellular bacteria, which find refuge from bactericidal devices by hiding within mammalian cells. Unfortunately, many therapeutics, such as vancomycin, do not possess membrane penetrating abilities to achieve efficacious eradication of bacteria at the subcellular level, allowing infections to persist. In an effort to target pathogens that thrive within mammalian cells, features of cell penetrating peptides (CPPs) and antimicrobial peptides (AMPs) were combined to develop a dual action antimicrobial CPP, cationic amphiphilic polyproline helices (CAPHs). CAPHs have proven to be an effective antimicrobial agent to combat an array of both Gram negative and Gram positive bacteria. </p> <p> </p> <p>Herein, to improve CAPHs activity, we have demonstrated how the incorporation of strategic modifications has resulted in increased cell uptake, alternative subcellular locations for CAPHs, and advanced antimicrobial potency. By simultaneously extending the helical length of CAPHs while incorporating different hydrophobic groups in place of the original isobutyl moiety that compose CAPHs we have created a <b>FL-P17-5R </b>series of peptides with five carbon aliphatic motifs: <b>Fl-P17-5B</b>, <b>Fl-P17-5C</b> and <b>Fl-P17-5L. </b>Through these modifications the peptides proved to be 2 to 5-fold more efficient in accumulating in macrophage cells than parent peptide Fl-P14LRR and where able to clear intracellular pathogenic bacteria, such as <i>Listeria</i>, from infected macrophages by 26 to 54%. </p> <p> </p> <p>In addition to making the <b>Fl-P17-5R</b> series of CAPHs to potentiate CAPHs activity, modifications to the cationic moiety of CAPHs were explored. By incorporating a new cationic monomer into the CAPHs sequence, a guanylated amino proline (GAP) residue, we produced <b>Fl-P14GAP</b>, a CAPHs peptide with an organized cationic charge display. This modification resulted in a 5-fold increase in cell uptake and a 2 to 16-fold decrease in minimum inhibitory concentration (MIC) values against strains of enteric and ESKAPE pathogens in comparison to Fl-P14LRR. <b>Fl-P14GAP</b> also executed superior clearance of intracellular pathogenic bacteria that resulted in the complete eradication of a drug resistant strain of <i>A. baumannii</i> from infected macrophage cells. Overall, our efforts with the <b>Fl-P17-5R</b> series of CAPHs and <b>Fl-P14GAP</b> have strengthened the therapeutic potential of CAPHs in the hopes of addressing the need for novel antibiotics with the propensity to eradicate intracellular pathogens.</p>

<p>Peptides, specifically cell-penetrating peptides (CPP), have become wonderful research tools due to their enhanced stability, solubility, and ease of synthesis. They have been used for a wide range of biomedical applications, from insecticides to biosensors and drug-delivery scaffolds. The work presented in this dissertation characterizes the biophysical properties of two different CPPs. The first is the cationic amphiphilic polyproline helix (CAPH) peptide, P14LRR. In addition to cell penetration, this CPP has demonstrated broad spectrum antibacterial properties. Fluorescence polarization (FP) and SEC-MALS were conducted to understand the dissociation constant (KD) and oligomerization effects of P14LRR with respect to its putative molecular target in Staphylococcus aureus (S. aureus). A biotinylated derivative of this peptide was also used as a drug-delivery scaffold to transport fluorescently conjugated streptavidin into mammalian cells. A second CPP, DAN13, was also developed as a biosensor for phosphoinositide lipids, specifically PI(4,5)P2. This was effected through careful calibration using stacked supported lipid bilayers (SSLB) in combination with total internal reflection fluorescence (TIRF) microscopy. This was then used to determine the absolute densities and spatial distribution of PIP2 in live KRas mutant cells.</p>

Von Beginn der flexiblen Automatisierung mit numerisch gesteuerten Werkzeugmaschinen und der zugehörigen Programmier-Software bis zum gegenwärtigen Entwicklungsstand (Industrie 4.0) wird die historische Entwicklung von 1960 bis 2020 aus der Position eines aktiven Mitgestalters dargestellt. Interessanterweise vollzogen sich die wesentlichen Entwicklungsetappen für die ersten dreißig Jahre parallel in beiden deutschen Staaten. Aus den Lehren des Zweiten Weltkrieges wurden im Rahmen der UNESCO zum friedlichen Informationsaustausch geeignete wissenschaftliche Organisationen gegründet: • IFIP (Internationale Föderation für Informationsprozesse, speziell Arbeitsgrupp CAM • CIRP (Internationale Akademie der Fertigungstechniker) Mit der Berufung und aktiven Mitarbeit in diesen Organisationen war eine Plattform für die deutsch-deutsche und darüber hinaus internationale Kooperation gegeben. Ein besonderer Schwerpunkt für den geordneten Informationsaustausch im Rahmen der gesamten dynamischen Entwicklung im Gebiet der Produktionsautomatisierung war dabei die im 3-Jahres-Rhythmus durchgeführte Konferenzserie PROLAMAT (Programming Languages for Machine Tools), gestartet 1969 in Rom. Im weiteren Verlauf wurde dieser Begriff viel breiter für das gesamte Gebiet der automatisierten Informationsverarbeitung und Fertigung erweitert. Ein besonderer Höhepunkt war dabei die erfolgreichste PROLAMAT-Konferenz 1988 in Dresden. Parallel dazu erfolgten an der TU Dresden Entwicklungen in Richtung CAD/CAM-Labor und später CIM-TT (CIM-Technologietransferzentrum). Damit war an der TU Dresden 1989/90 ein Entwicklungsstand gegeben, der unmittelbar zu gemeinsamen deutsch-deutschen und internationalen EU-Projekten genutzt werden konnte. Dieses hohe Entwicklungsniveau wurde zur offiziellen Eröffnung des CIM-TT-Zentrums in den Eröffnungsreferaten durch den damaligen Wissenschaftsminister Dr. Riesenhuber und Ministerpräsident Prof. Biedenkopf gewürdigt. Durch die zum gleichen Zeitpunkt verfügte veränderte Nutzung des für das CIM-TT im Aufbau befindliche Gebäude durch die neugegründete Juristische Fakultät wurde der erfolgreich vorbereitete Weg verhindert. Unabhängig davon blieb meine fachliche Orientierung mit den gravierenden Weiterentwicklungen eng verbunden. Dazu trug das Sabbatical-Jahr in Norwegen und den USA 1992 maßgeblich bei. Mit dem Forschungsaufenthalt war die Entscheidungsvorbereitung für die vorgesehene Groß-Investition für das neueste generative Verfahren verbunden. Gleichzeitig mit dem fundierten Nachweis der bestgeeigneten sog. Rapid-Prototyping-Anlage vom deutschen Anbieter EOS München war die TU Dresden auf diesem neuen High-Tech-Gebiet 1992 in einer anerkannten Spitzenposition. Mit meiner Publikation eines der ersten Fachbücher im Gebiet Advanced Prototyping (jetzt Additiv Manufacturing) war darüber hinaus eine gute Basis für weitere innovative Aktivitäten gegeben Dazu gehört die Gründung einer High-Tech-Firma (SFM - Schnelle Fertigung von Modellen) mit bemerkenswerten beispielgebenden Ergebnissen. Hervorgehoben soll die zwanzigjährige aktive Kooperation mit der Universität Stellenbosch (RSA - Republik Südafrika), die unter anderem mit meiner Berufung zum Extraordinary Professor im Jahr 2003 verbunden ist. Mit der Eröffnung eines Technologie-Zentrums nach dem Vorbild des ursprünglichen CIM TT -Zentrums der TU Dresden konnte für Südafrika ein wertvoller Beitrag geleistet werden. Das gesamte Lebenswerk ist gekennzeichnet durch die Entwicklungsschritte von der Mathematisierung über die Algorithmierung bis hin zur Programmierung vielfältiger technologischer Sachverhalte. Die Ergebnisse sind in einer Anzahl von persönlichen Fachbüchern (z.T. übersetzt in das Russische und Ungarische) wie auch Konferenzberichten und mehr als 200 Veröffentlichungen (deutsch und englisch) dokumentiert.