TY - THES A1 - Torzewska, Agata T1 - Method development for quantification of prescription drugs Fluphenazine and Flupentixol in serum. N2 - The LVR clinic in Viersen belongs to a group of nine psychiatric clinics in NRW, Germany. In the pharmacy laboratory, the therapeutic drug monitoring is offered for antiepileptic drugs (AED´s), neuroleptics and antidepressants. Fluphenazine and Flupentixol are used to relieve the symptoms of schizophrenia and other similar mental health problems. The purpose of this work is the development and validation of methods for quantitation of the neuroleptics Fluphenazine and Flupentixol, contained in human serum. Therapeutic drug monitoring (TDM) is very important for a patient-matched therapy. TDM is applied to handle and direct the pharmacotherapy because of the pharmacokinetic differences between different individuals. The developed methods should be afterwards introduced into the daily routine of the laboratory. The analytical device used for this purpose is a Liquid Chromatography with a Mass Spectrometer (LCMS), with a Biphenyl Accucore separation column which is also used for other neuroleptics quantised. For Fluphenazine, an already existing in the laboratory sample precipitation method was used. Also, an already applied for other similar neuroleptics LCMS method, called “Biphenyl03_new” (lasting for 11 minutes, with a high gradient between the elution agents) was taken on, and resulted in a good linearity. In case of Flupentixol, a completely new method had to be established, in order to achieve a baseline separation of cis-Z-Flupentixol and trans-E-Flupentixol. For the sample preparation, a solid phase extraction method was developed. The established methods were then applied for quantitation of patient samples, and the stability of the measurements was proved. Also, the accuracy and precision of obtained results was tested in the Proficiency Test. Limit of quantitation achieved for Fluphenazine was 0,5ng/ml, whereas for Flupentixol the LOQ was 0,25ng/ml. The Limit of detection found was 0,05ng/ml for both substances. The development was successful for both analytes and the established methods were integrated into the daily routine of the laboratory. Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:1383-opus4-12715 ER - TY - THES A1 - Birkenbach, Gesa T1 - Quantitative real-time PCR Detection of Yersinia ruckeri for Application in Aquaculture N2 - To detect pathogen infections in aquaculture early and reduce their impact, a photonic biosensing chip is being developed in the PHOTO-SENS project, to enable fast and easy routine testing in aquaculture facilities. One target bacterial pathogen is Yersinia ruckeri, the causative agent of enteric redmouth disease. Detection is based on short DNA sequences (probes), that are complementary to the DNA of their target pathogen. In this work, a Yersinia ruckeri-specific probe was designed together with a set of qPCR primers. The probe and primers were tested in qPCR experiments, including quantifi- cation of extracted DNA, specificity and sensitivity testing, and detection of bacterial DNA from highly diluted samples to mimic the detection of environmental DNA. Three different standard series were created for the quantification of unknown samples. The influence of fish tissue on detection was tested, and the efficiency of Chelex-DNA ex- tractions were compared to peqGOLD Tissue DNA Mini Kit extractions. During quantification experiments with a first standard series (Y1P), the quantification output of most samples was 12–175 times higher than expected. The lower limit of detection was at 100 copies based on that standard, with an efficiency of 88%. The discrepancy was reduced with a gBlocks standard (limit of detection: 5 copies; effi- ciency: 92%). A CFU standard provided quantification results in reasonable agreement with expected CFU starting quantities (limit of detection: 1 CFU; efficiency: 106%). De- tection of diluted samples after filtration was possible. Chelex DNA extractions yielded higher amounts of detectable DNA compared to a DNA extraction kit. Less bacterial DNA was extracted in bacterial culture samples containing fish tissue than in samples that did not contain fish tissue. Higher-than-expected quantification outputs were attributed to flaws in the creation of a first standard series as well as detectable DNA from dead cells or clusters that were not accounted for in the expected starting quantities. The gBlocks standard was as- sumed to be most reliable for quantification of exact copy numbers, while the CFU standard produced results that could be accounted for in CFU counts. As these chal- lenges concerned the standard series’ and not the probe itself, it was concluded that the probe was suitable for application in the PHOTO-SENS project for detection of Yersinia ruckeri. KW - Quantitative real-time PCR (qPCR) KW - Yersinia ruckeri KW - DNA quantification KW - Hydrolysis probe KW - Aquaculture Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:1383-opus4-16361 ER - TY - THES A1 - Waller, Ole T1 - Evaluating the potential of mRNA vaccines for human rabies N2 - To this date, rabies is still one of the most fatal diseases worldwide and generates at least 59,000 deaths per year. Although current vaccines are safe and effective, RNA technology proposes improved vaccines that are potentially able to facilitate eradication of rabies until 2030. In this review, RNA vaccine candidates are evaluated for their potential to be effective against rabies and to be implemented in immunization programs. To understand the relevance of disease prevention, rabies virus and its function in the body is clarified. RNA technology is explained to comprehend its mode of action in the human immune system and its potential to be a vaccine platform. Comparison to existing vaccines showed advantages in several characteristics that are relevant for vaccine approval. RNA vaccines scored better in efficacy, stability, production, and dosing, but there is still room for improvements in terms of safety and tolerability. Theoretical potential is proven, but practical translation of RNA vaccines into clinical candidates requires more attention, as safety and tolerability are the leading limiting factor for approval of any new drug or vaccine. Producing a safe, stable and effective RNA vaccine does not necessarily ensure less rabies deaths, because rabies control strategies heavily rely on proper funding and availability in these rabies endemic region. Nonetheless, RNA vaccine technology offers potential solutions for the elimination of rabies. Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:1383-opus4-16737 ER - TY - THES A1 - Hawas, Takwa T1 - Chancen und Risiken der CAR-T-Zellen in der onkologischen Immuntherapie N2 - Bei der CAR-T-Zell-Therapie erhalten Patient*innen nach einer Leukapherese autologer T-Lymphozyten ihre eigenen T-Zellen zurück, die vor der Behandlung mittels viraler Vektoren die chimären Antigenrezeptoren, engl. Chimeric Antigen Receptor (CAR), transduziert bekommen. Bereits Stunden nach der Zellinfusion und in den folgenden Wochen expandieren die CAR-T-Zellen und sind in der Lage, das Antigen spezifisch zu binden und die Tumorzellen zu zerstören. CAR-T-Zellen haben sich in Therapien gegen hämatologische Tumore als spektakulär wirksam erwiesen. Doch die vielversprechende Behandlung hat ihre Grenzen und ihren Preis. Drei große Herausforderungen begrenzen derzeit ihren Erfolg im Bereich solider Tumore: Antigenerkennung, Migration und Überleben in der Tumormikroumgebung. CAR-T-Zell-Therapien zählen auch mit Kosten von etwa 300 Tausend Euro immer noch zu den teuersten Verfahren in der Medizin. Eine intensivere Forschung im Bereich der Dysfunktion des Immunsystems in der Tumormikroumgebung stellt ein notwendiger Schritt zur Überwindung von Hürden bei der Behandlung von soliden Tumoren mit CAR-T-Zellen dar. Angemessene finanzielle sowie gesetzliche Anreize und die Einführung optimierter Herstellungsprozesse, multidisziplinärer und prädiktiver Ansätze und zeitgemäßer Tools der Biotechnologie und Bioinformatik in den klinischen Alltag zur prädiktiven Krebsmodellierung sind ebenso wichtige Grundvoraussetzungen für eine wirtschaftliche Ausweitung des Einsatzes von CAR-T-Zellen. Anderenfalls würden Gesundheitssysteme die Kosten sowie die Komplexität nicht mehr stemmen können und diese lebensrettende CAR-T-Zell-Therapien würden außerhalb der Reichweite der Patienten bleiben, die es wirklich brauchen. KW - CART KW - CAR-T-Zellen KW - CAR-T-cells KW - Immuntherapien KW - CAR-T Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:1383-opus4-16513 ER - TY - THES A1 - Abdelazim, Ahmed T1 - A Structured Classification and Evaluation of Current and Future Treatments for Systemic Lupus Erythematosus N2 - Systemic Lupus Erythematosus (SLE) is a heterogeneous autoimmune disease that has a variety of manifestations and is therefore difficult to find a treatment for. Several advances have occurred in the field of SLE drug discovery, especially since the publishing of the first Genome Wide-Associated Studies in 2005. More than one hundred loci were found to be involved in the disease. Subsequently, the treatments were aimed towards tackling the pathways that were found after the uses of modern genetic techniques. Historically, the FDA approved only two drugs for treatment until the approval of Belimumab in 2011. Ten years later, two more drugs were approved in the same year, with few more drugs expected to receive approval in the near future. This study aims to evaluate the efficacy of these future drugs in addition to the ones considered to be milestones in the treatment of SLE, finding Anifrolumab to be the most promising drug currently on the market for treatment of moderate to severe SLE and Obinutuzumab to be the drug to watch. Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:1383-opus4-16960 ER - TY - THES A1 - Chentsova, Olga T1 - MESENCHYMAL STEM CELLS (MSC) IN BREAST CANCER TREATMENT: ADVANTAGES AND CHALLENGES OVER TRADITIONAL THERAPIES N2 - Breast cancer is one of the most prevalent and deadly malignancies in women globally, due to its aggressive behavior and poor response to traditional treatments. Despite the significant improvements in understanding of tumor therapies, efforts to treat patients with advanced cancer are still hampered by the size and heterogeneity of the tumor burden and the development of drug resistance. Therefore, the need for novel therapeutic strategies is essential. Mesenchymal stem cells (MSCs) have been studied for a long time with encouraging results in the treatment of metastatic cancers, including breast cancer. The discovery of MSCs' therapeutic abilities to inhibit the growth of cancer, cause cell death, and trigger immune responses served as a catalyst for the development of bioengineered MSCs. The use of MSCs in cell therapy is the subject of numerous investigations now. This thesis aimed to summarize the current knowledge on traditional breast cancer treatments and compare them to novel approaches based on MSCs usage. To this end, scientific publications retrieved from databases such as PubMed were selected considering specific inclusion/exclusion criteria and were extensively analyzed. Results obtained underline the potential applications of MSCs for breast cancer treatment, such as using MSCs as carriers of oncolytic viruses, suicide genes, and anticancer medicines to malignancies. Furthermore, challenges and limitations of several MSCs based treatments and MSCs control were evaluated and discussed based on ongoing clinical trials. KW - MSC KW - Mesenchymal stem cells KW - Traditional cancer therapies breast cancer KW - BC KW - Treatment Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:1383-opus4-16927 ER -