The effects of washing a collagen sample prior to TEM examination

Transmission electron microscopy (TEM) is an important analysis technique to visualize (bio)macromolecules and their assemblies, including collagen fibers. Many protocols for TEM sample preparation of collagen involve one or more washing steps to remove excess salts from the dispersion that could hamper analysis when dried on a TEM grid. Such protocols are not standardized and washing times as well as washing solvents vary from procedure to procedure, with each research group typically having their own protocol. Here, we investigate the influence of washing with water, ethanol, but also methanol and 2-propanol, for both mineralized and unmineralized collagen samples via a protocol based on centrifugation. Washing with water maintains the hydrated collagen structure and the characteristic banding pattern can be clearly observed. Conversely, washing with ethanol results in dehydration of the fibrils, often leading to aggregation of the fibers and a less obvious banding pattern, already within 1 min of ethanol exposure. As we show, this process is fully reversible. Similar observations were made for methanol and propanol. Based on these results, a standardized washing protocol for collagenous samples is proposed.     

Evolution of MicroRNA Biogenesis Genes in the Sterlet (Acipenser ruthenus) and Other Polyploid Vertebrates

MicroRNAs play a crucial role in eukaryotic gene regulation. For a long time, only little was known about microRNA-based gene regulatory mechanisms in polyploid animal genomes due to difficulties of polyploid genome assembly. However, in recent years, several polyploid genomes of fish, amphibian, and even invertebrate species have been sequenced and assembled. Here we investigated several key microRNA-associated genes in the recently sequenced sterlet (Acipenser ruthenus) genome, whose lineage has undergone a whole genome duplication around 180 MYA. We show that two paralogs of drosha, dgcr8, xpo1, and xpo5 as well as most ago genes have been retained after the acipenserid-specific whole genome duplication, while ago1 and ago3 genes have lost one paralog. While most diploid vertebrates possess only a single copy of dicer1, we strikingly found four paralogs of this gene in the sterlet genome, derived from a tandem segmental duplication that occurred prior to the last whole genome duplication. ago1,3,4 and exportins1,5 look to be prone to additional segment duplications producing up to four-five paralog copies in ray-finned fishes. We demonstrate for the first time exon microsatellite amplification in the acipenserid drosha2 gene, resulting in a highly variable protein product, which may indicate sub- or neofunctionalization. Paralogous copies of most microRNA metabolism genes exhibit different expression profiles in various tissues and remain functional despite the rediploidization process. Subfunctionalization of microRNA processing gene paralogs may be beneficial for different pathways of microRNA metabolism. Genetic variability of microRNA processing genes may represent a substrate for natural selection, and, by increasing genetic plasticity, could facilitate adaptations to changing environments.     

Ärztliche Schweigepflicht bei Kindeswohlgefährdung

Definition of the problem Medical confidentiality, which is one of the main pillars of medical ethics, has taken a decisive role in the latest discussions about the improvement of child protection laws. Inexplicit and plurivalent arrangements and procedures of how to legally handle child abuse cases when persons with legal custody refuse to take further support are seen as an obstacle for effective child protection. Arguments Non-specific legal regulations lead to task uncertainty and hesitation on the side of health care professionals, which can result in the delay or even prevention of necessary actions. Therefore, new laws on the federal level have been passed in order to establish certainty with regard to professional secrecy and in order to foster networking among the actors in the field of child protection. Conclusion The paper analyses and discusses these legal attempts at strengthening child protection. Although some of the federal laws have the potential of clarification, the great number of laws and their explication restricts their impact.     

Bolus ingestion of white and green tea increases the concentration of several flavan‐3‐ols in plasma,but does not affect markers of oxidative stress in healthy non‐smokers

White tea (WT) is rich in flavan‐3‐ols as green tea (GT) and might provide health protective effects due to the strong antioxidant properties of flavan‐3‐ols. Since intervention studies with WT are lacking, we evaluated the effects of WT consumption on antioxidant status, antioxidant capacity and biomarkers of oxidative stress compared to water and GT. After an overnight fast, 70 healthy non‐smokers were randomized to consume 600 mL of WT, GT or water (control). Plasma (epi‐)catechin and epi(gallo)catechingallate, antioxidant capacity (Folin assay, trolox equivalent antioxidant capacity test), 8‐iso‐prostaglandin F_2α, ascorbic acid and uric acid were determined before and several times within 8 h after consumption. DNA strand breaks were measured in vivo and ex vivo (H₂O₂ stimulation) in leukocytes. Plasma flavan‐3‐ols significantly increased after WT and GT ingestion. Trolox equivalent antioxidant capacity was lower after 5 h in controls versus WT (p=0.031) and GT (p=0.005). Folin‐Ciocalteu reducing capacity, ascorbic and uric acid as well as markers of oxidative stress (8‐iso‐prostaglandin‐F_2α, DNA strand breaks) were not affected by the beverages. A short‐term increase of catechins does not change plasma antioxidant capacity in healthy subjects. Conclusions with respect to health protective effects of WT and GT on the basis of these biomarkers can, thus, not be drawn.     

Intrinsic Seebeck Coefficient of Quantum Dots

The Seebeck coefficient S is an important performance characteristic of thermoelectric materials. In this paper we establish the fact that quantum dots and single-electron tunneling devices with narrow, well-spaced energy levels and sharp transmission resonances have a Seebeck coefficient independent of material parameters. By employing a delta function for the transmission resonances we arrive at an intrinsic expression for S in terms of the fundamental electronic charge e. We further confirm the validity of our result in the case of a transmission resonance with finite width.     

Oligophenylenevinylenes in Spatially Confined Nanochannels: Monitoring Intermolecular Interactions by UV/Vis and Raman Spectroscopy

Perhydrotriphenylene‐based channel‐forming inclusion compounds (ICs) and thin films made of polyphenylenevinylene (PPV)‐type oligomers with terminal alkoxy groups are investigated and compared in a combined experimental and theoretical approach. Interchromophore interactions and host‐guest interactions are elucidated by UV/Vis and Raman spectroscopy. The impact of the local environment of the chromophore on the optical and photophysical properties is discussed in light of quantum‐chemical calculations. In stark contrast to thin films where preferential side‐by‐side orientation leads to quenching of photoluminescence (PL) via non‐emissive traps, the ICs are found to be attractive materials for opto‐electronic applications: they offer high chromophore concentrations, but at the same time behave as quasi‐isolated entities of tightly packed, well‐oriented objects with high PL quantum yields and the possibility of color tuning.     

Über die Anwendung von Ionensensitiven Feldeffekttransistoren zur Untersuchung von Korrosionsvorgängen

On the application of ion sensitive field-effect transistors for the investigation of corrosion phenomena Ion sensitive field-effect transistors (ISFET) are chemical sensors based on semiconductor technology. They exhibit small dimensions, a plane measuring surface and have short response times. Thus, they are very suitable for the determination of local changes of the pH value at electrochemical corrosion studies. The described in this paper experimental set-up allows in situ measurements of the pH value by means of ISFET sensors in amounts of liquids below 1 mm³; the active surface area of the material under test may be below 1 mm². In model tests preferably changes of the pH value of dist. water due to the corrosion of steel St 38 have been investigated. After an initial short-time rise caused by oxygen reduction a decay of the pH value to pH = 3 ?4 was obseved. Furthermore, ISFET sensors have been used as a potential probe for electrochemical polarization measurements in low conductive media. It was shown, that the dynamic behaviour of potentiostatic control circuits may be influenced favourably by the low output resistance of the ISFET.