Zur Vergiftungsproblematik in der Region Thüringen von 1820 bis 1900 - Fallbeschreibungen, Noxen, Intoxikationsursachen, Gesetze und Verordnungen, Nachweise

Die Dissertation basiert auf einer umfassenden Recherche in den Thüringer Staatsarchiven. Für den Zeitraum von 1820 bis 1900 konnten für die Thüringer Region 89 valide Intoxikationsfälle gefunden werden. Die Intoxikationen waren meist akzidenteller Natur, betrafen aber auch Fälle aus dem Medizinal-Bereich und der Gewerbetoxikologie. Ferner konnten Giftmorde und suizidale Vergiftung nachgewiesen werden. Das Spektrum an Giften umfasste anorganische, organische und pflanzliche Noxen. Arsenik wurde am häufigsten eingesetzt, wobei Vergiftungen mit dieser Noxe meist tödlich verliefen. Die typische Intoxikation kann als akute Vergiftung mit peroraler Aufnahme charakterisiert werden. Ergänzend wurden bestehende Gesetze zum Umgang und Handel mit Giften sowie der Giftnachweis untersucht.von Michael Pateng

Antisense-Regulation der Genexpression von Streptococcus pyogenes in Virulenz und Therapie

Streptococcus pyogenes ist ein strikt humanpathogenes Bakterium, das weltweit eine hohe Belastung der menschlichen Gesundheit und der Gesundheitssysteme darstellt. In dieser Arbeit lag der Fokus auf der Untersuchung der Antisense-Regulation der Genexpression. Zum einen wurden kleine regulatorische RNAs identifiziert und ihre Bedeutung für die Genregulation in S. pyogenes an zwei Beispielen untersucht. Zum anderen wurde ein Antisense-basierter Therapieansatz mit Carrierpeptid-gekoppelten Peptid-Nukleinsäuren für S. pyogenes etabliert

Oxacillin sensitization of methicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus pseudintermedius by antisense peptide nucleic acids in vitro

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.BACKGROUND: Antibiotic resistance genes can be targeted by antisense agents, which can reduce their expression and thus restore cellular susceptibility to existing antibiotics. Antisense inhibitors can be gene and pathogen specific, or designed to inhibit a group of bacteria having conserved sequences within resistance genes. Here, we aimed to develop antisense peptide nucleic acids (PNAs) that could be used to effectively restore susceptibility to β-lactams in methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP). RESULTS: Antisense PNAs specific for conserved regions of the mobilisable gene mecA, and the growth essential gene, ftsZ, were designed. Clinical MRSA and MRSP strains of high oxacillin resistance were treated with PNAs and assayed for reduction in colony forming units on oxacillin plates, reduction in target gene mRNA levels, and cell size. Anti-mecA PNA at 7.5 and 2.5 μM reduced mecA mRNA in MRSA and MRSP (p < 0.05). At these PNA concentrations, 66 % of MRSA and 92 % of MRSP cells were killed by oxacillin (p < 0.01). Anti-ftsZ PNA at 7.5 and 2.5 μM reduced ftsZ mRNA in MRSA and MRSP, respectively (p ≤ 0.05). At these PNA concentrations, 86 % of MRSA cells and 95 % of MRSP cells were killed by oxacillin (p < 0.05). Anti-ftsZ PNAs resulted in swelling of bacterial cells. Scrambled PNA controls did not affect MRSA but sensitized MRSP moderately to oxacillin without affecting mRNA levels. CONCLUSIONS: The antisense PNAs effects observed provide in vitro proof of concept that this approach can be used to reverse β-lactam resistance in staphylococci. Further studies are warranted as clinical treatment alternatives are needed.Peer reviewedFinal Published versio

Promoter analysis by saturation mutagenesis

Gene expression and regulation are mediated by DNA sequences, in most instances, directly upstream to the coding sequences by recruiting transcription factors, regulators, and a RNA polymerase in a spatially defined fashion. Few nucleotides within a promoter make contact with the bound proteins. The minimal set of nucleotides that can recruit a protein factor is called a cis-acting element. This article addresses a powerful mutagenesis strategy that can be employed to define cis-acting elements at a molecular level. Technical details including primer design, saturation mutagenesis, construction of promoter libraries, phenotypic analysis, data analysis, and interpretation are discussed

Analysis of differential DNA damage in the mitochondrial genome employing a semi-long run real-time PCR approach

The maintenance of the mitochondrial genomic integrity is a prerequisite for proper mitochondrial function. Due to the high concentration of reactive oxygen species (ROS) generated by the oxidative phosphorylation pathway, the mitochondrial genome is highly exposed to oxidative stress leading to mitochondrial DNA injury. Accordingly, mitochondrial DNA damage was shown to be associated with ageing as well as with numerous human diseases including neurodegenerative disorders and cancer. To date, several methods have been described to detect damaged mitochondrial DNA, but those techniques are semi-quantitative and often require high amounts of genomic input DNA. We developed a rapid and quantitative method to evaluate the relative levels of damage in mitochondrial DNA by using the real time-PCR amplification of mitochondrial DNA fragments of different lengths. We investigated mitochondrial DNA damage in SH-SY5Y human neuroblastoma cells exposed to hydrogen peroxide or stressed by over-expression of the tyrosinase gene. In the past, there has been speculation about a variable vulnerability to oxidative stress along the mitochondrial genome. Our results indicate the existence of at least one mitochondrial DNA hot spot, namely the D-Loop, being more prone to ROS-derived damage

Acetylation increases access of remodelling complexes to their nucleosome targets to enhance initiation of V(D)J recombination

Targeted chromatin remodelling is essential for many nuclear processes, including the regulation of V(D)J recombination. ATP-dependent nucleosome remodelling complexes are important players in this process whose activity must be tightly regulated. We show here that histone acetylation regulates nucleosome remodelling complex activity to boost RAG cutting during the initiation of V(D)J recombination. RAG cutting requires nucleosome mobilization from recombination signal sequences. Histone acetylation does not stimulate nucleosome mobilization per se by CHRAC, ACF or their catalytic subunit, ISWI. Instead, we find the more open structure of acetylated chromatin regulates the ability of nucleosome remodelling complexes to access their nucleosome templates. We also find that bromodomain/acetylated histone tail interactions can contribute to this targeting at limited concentrations of remodelling complex. We therefore propose that the changes in higher order chromatin structure associated with histone acetylation contribute to the correct targeting of nucleosome remodelling complexes and this is a novel way in which histone acetylation can modulate remodelling complex activity

Regulation of Translation in Haloarchaea: 5′- and 3′-UTRs Are Essential and Have to Functionally Interact In Vivo

Recently a first genome-wide analysis of translational regulation using prokaryotic species had been performed which revealed that regulation of translational efficiency plays an important role in haloarchaea. In fact, the fractions of genes under differential growth phase-dependent translational control in the two species Halobacterium salinarum and Haloferax volcanii were as high as in eukaryotes. However, nothing is known about the mechanisms of translational regulation in archaea. Therefore, two genes exhibiting opposing directions of regulation were selected to unravel the importance of untranslated regions (UTRs) for differential translational control in vivo

Evaluating FRP/Concrete Bond Using Infrared Thermography

Damaged concrete structures are often reinforced with fiber-reinforced polymer (FRP) composites. Although FRP composites are known to greatly increase the strength of these structures, enhancement is dependent upon the bond between the concrete and the FRP composite. The current method of evaluating that bond is known as the pull-off test in which a circle is drilled through the FRP and concrete, a steel disk is then affixed to the FRP composite and pulled off. The layer that fails is the weakest layer. If the bond is correct the test will fail in the concrete. This test is very destructive and only tests the one circular section; infrared thermal imaging can test the entire area and is not harmful to the test specimen. This research focused on using infrared thermal imaging in conjunction with the pull-off test to better evaluate the bond and prove the legitimacy of thermal imaging. Twenty-four small concrete blocks were built, FRP was applied, images were taken and processed, and 104 holes were drilled and are ready to be tested in the fall. Results will come shortly