Advanced Mutasynthesis Studies on the Natural α‐Pyrone Antibiotic Myxopyronin from Myxococcus fulvus

Myxopyronin is a natural α‐pyrone antibiotic from the soil bacterium Myxococcus fulvus Mx f50. Myxopyronin inhibits bacterial RNA polymerase (RNAP) by binding to a part of the enzyme not targeted by the clinically used rifamycins. This mode of action makes myxopyronins promising molecules for the development of novel broad‐spectrum antibacterials. We describe the derivatization of myxopyronins by an advanced mutasynthesis approach as a first step towards this goal. Site‐directed mutagenesis of the biosynthetic machinery was used to block myxopyronin biosynthesis at different stages. The resulting mutants were fed with diverse precursors that mimic the biosynthetic intermediates to restore production. Mutasynthon incorporation and production of novel myxopyronin derivatives were analyzed by HPLC‐MS/MS. This work sets the stage for accessing numerous myxopyronin derivatives, thus significantly expanding the chemical space of f α‐pyrone antibiotics.     

Functionalized gold nanoparticles: a detailed in vivo multimodal microscopic brain distribution study

In the present study, the in vivo distribution of polyelectrolyte multilayer coated gold nanoparticles is shown, starting from the living animal down to cellular level. The coating was designed with functional moieties to serve as a potential nano drug for prion disease. With near infrared time-domain imaging we followed the biodistribution in mice up to 7 days after intravenous injection of the nanoparticles. The peak concentration in the head of mice was detected between 19 and 24 h. The precise particle distribution in the brain was studied ex vivo by X-ray microtomography, confocal laser and fluorescence microscopy. We found that the particles mainly accumulate in the hippocampus, thalamus, hypothalamus, and the cerebral cortex.     

Fiber scaffolds of polysialic acid via electrospinning for peripheral nerve regeneration

Fiber scaffolds of bioactive polysialic acid have been prepared via electrospinning for peripheral nerve regeneration. The diameter, morphology and alignment of fibers in scaffolds were adjusted by variation of electrospinning parameters, which are decisive for the cell-scaffold interaction. Due to the high water solubility of polysialic acid (poly-α-2,8-N-acetylneuraminic acid) a photoactive derivative (poly-α-2,8-N-pentenoylneuraminic acid) was used to obtain stable fiber scaffolds in water by photochemical crosslinking. At the optimized fiber scaffolds good cell viability and directed cell proliferation along the fibers was achieved by cell tests with immortalized Schwann cells.     

Verhalten von Mauerwerksbauten unter Erdbebeneinwirkung: Auswertung der Schäden des Albstadt‐Erdbebens vom 3. September 1978

Numerische Untersuchungen zum Verhalten von Mauerwerksbauten unter Erdbebeneinwirkung führen oft zu pessimistischen Schadensprognosen, welche im Gegensatz zu den Beobachtungen stehen. Um diesen Widerspruch aufzulösen, werden erfahrungsbasierte Verletzbarkeitsfunktionen für typische Mauerwerksgebäude auf der Grundlage der durch das Erdbeben vom 3. September 1978 bei Albstadt (mit einer Lokalmagnitude M_L von 5,7 und Epizentralintensität I₀ = VII—VIII) verursachten Schäden entwickelt. Anhand des Bauwerksbestandes von 1978 erfolgt die Einordnung der beobachteten Schäden in Schadensgrade auf Basis der European Macroseismic Scale EMS‐98 [1]. Die für den Bauwerksbestand repräsentativen Bauweisen werden herausgearbeitet; für die vorherrschenden Gebäude aus unbewehrtem Mauerwerk ist eine weitere Unterscheidung nach Baualter, Geschosszahl und phänomenologischen Gesichtspunkten möglich. Vulnerability of masonry structures under seismic action: Damage analysis of the September 3, 1978 Albstadt earthquake. Numerical studies of the earthquake behavior of masonry buildings in Central Europe based on national building codes acc. to the Eurocode 8 lead to pessimistic damage prognoses, which are in contradiction to the observed behavior. In order to eliminate this discrepancy realistic experience‐based vulnerability and displacement functions for typical masonry constructions are developed. Because of the rather limited number of earthquake damage observations, the Magnitude M_L 5.7 Albstadt earthquake from September 3, 1978 (intensity VII—VIII) in South Germany also based on its excellent documentation is reconstructed with the building stock existing at that time. The prevailing building types and for these the characteristic damage cases are investigated in close cooperation with the local authorities. The presented unreinforced masonry structures are divided by year of construction, number of storeys and phenomenological aspects.     

A novel family of solid basic catalysts obtained by nitridation of crystalline microporous aluminosilicates and aluminophosphates

Novel basic catalysts are obtained by ammonia treatment of crystalline, microporous aluminosilicates (zeolites) and aluminophosphates at temperatures above 800°C. The resulting materials are active catalysts in the Knoevenagel condensation of benzaldehyde with malononitrile, presumably due to the presence of nitrogen-containing species bound to the crystalline framework. While nitridation of zeolite NaY at temperatures around or below 800°C does not result in an increase of the catalytic activity, ammonia treatment at 850–875°C produces a significantly more active material. Further typical experimental results are presented which suggest that the activity gain seems to depend not only on the temperature of ammonia treatment but also on the structure and the chemical composition of the parent material. The novel microporous catalysts with their reasonable base strength offer the principle possibility to perform base catalyzed reactions in a shape selective manner.     

Automated DNA preparation from maize tissues and food samples suitable for real-time PCR detection of native genes

There is an increasing need for high-throughput analyses of plants and food samples for the presence of specific DNA sequences, e.g. transgenic contaminations. We developed and optimized conditions for the automated isolation of DNA from several maize tissues and various edibles containing maize using the MagNA Pure LC system (Roche Applied Science). Our results show that the system provided is capable of isolating DNA from any tested source. Quantification of an endogenous gene by LightCycler real-time PCR revealed that the DNA is suitable in quality and quantity for multiple PCR analyses.     

Out of Context: NMDA Receptor Antagonism in the Avian 'Prefrontal Cortex' Impairs Context Processing in a Conditional Discrimination Task.

Processing of context information is implicated in prefrontal functions as response selection or attention. N-methyl-D-aspartate (NMDA) receptors in the mammalian prefrontal cortex (PFC) and in the nidopallium caudolaterale (NCL) of birds, the avian functional equivalent of the PFC, are involved in learning, which also requires processing of context. The authors investigated the role of NMDA receptors in the pigeon (Columba livia) NCL for context processing and response selection in a simultaneous-matching-to-sample task with 2 trial types, requiring either processing of context information, delivered by a conditional stimulus (context dependent), or only recall of a stimulus-response association (fixed response). The competitive NMDA antagonist DL-2-amino-5-phosphonovaleric acid impaired performance only in context-dependent trials. Therefore, NMDA receptors in the avian PFC participate in response selection requiring context processing rather than in response selection per se. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

\upalpha 5\upbeta 1-integrin and MT1-MMP promote tumor cell migration in 2D but not in 3D fibronectin microenvironments

Cell migration is a crucial event for physiological processes, such as embryonic development and wound healing, as well as for pathological processes, such as cancer dissemination and metastasis formation. Cancer cell migration is a result of the concerted action of matrix metalloproteinases (MMPs), expressed by cancer cells to degrade the surrounding matrix, and integrins, the transmembrane receptors responsible for cell binding to matrix proteins. While it is known that cell-microenvironment interactions are essential for migration, the role of the physical state of such interactions remains still unclear. In this study we investigated human fibrosarcoma cell migration in two-dimensional (2D) and three-dimensional (3D) fibronectin (FN) microenvironments. By using antibody blocking approach and cell-binding site mutation, we determined that $\upalpha _{5}\upbeta _{1}$ -integrin is the main mediator of fibrosarcoma cell migration in 2D FN, whereas in 3D fibrillar FN, the binding of $\upalpha _{5}\upbeta _{1}$ - and $\upalpha _\mathrm{v}\upbeta _{3}$ -integrins is not necessary for cell movement in the fibrillar network. Furthermore, while the general inhibition of MMPs with GM6001 has no effect on cell migration in both 2D and 3D FN matrices, we observed opposing effect after targeted silencing of a membrane-bound MMP, namely MT1-MMP. In 2D fibronectin, silencing of MT1-MMP results in decreased migration speed and loss of directionality, whereas in 3D FN matrices, cell migration speed is increased and integrin-mediated signaling for actin dynamics is promoted. Our results suggest that the fibrillar nature of the matrix governs the migratory behavior of fibrosarcoma cells. Therefore, to hinder migration and dissemination of diseased cells, matrix molecules should be directly targeted, rather than specific subtypes of receptors at the cell membrane.     

Encapsulation of naked dies for bulk silicon etching with TMAH

Because of the increasing number of metal levels within a semiconductor device and the ongoing transition to new package types failure analysis from the back side of a die becomes necessary. A useful chemical for bulk silicon removal is tetra-methyl-ammonium-hydroxide (TMAH). Because of the aggressive etch conditions, the edges of naked dies have to be protected against destruction by encapsulation into an appropriate molding compound. Different encapsulation materials were evaluated to identify a useful molding compound and an encapsulation method for bulk silicon removal of naked dies using TMAH. Problems as well as results are described in this paper.