Construction of an active humidity regulation setup for NMR/MRI-Observation and simulation of the controlled evaporation of sessile water droplets.

Controlling and improving processes like for example the production of organic semiconductors via printing depends on understanding the interplay of wetting and evaporation of complex fluids. Therefore, examination of the time dependent composition of complex fluid droplets during wetting or evaporation is of interest. The evaporation rate of sessile droplets containing largely water depends on the vapor pressures of the individual components and on the humidity (or partial pressure) of the surrounding gas phase. Hence, for a complete picture of an evaporation process and the comparability of the results of different measurements, it is essential to measure and control the humidity and temperature in the measurement compartment. Accordingly, climate chambers are available in different scales to fit a variety of techniques like contact angle goniometry to obtain results in a controlled atmosphere. We recently reported the application of MRI (Magnetic Resonance Imaging) and spatially resolved NMR (Nuclear Magnetic Resonance) spectroscopy for the examination of the evaporation of sessile droplets on surfaces in 10 mm NMR tubes. These are considered to be closed compartments. Here, we present an apparatus to a) measure and b) control the relative humidity within the sample compartment of the NMR setup by introducing preconditioned gas into the NMR tube. We monitored the evaporation of water droplets using RARE images and compared the volume decay with a) a simple diffusive evaporation model and b) with detailed FEM (finite element numerical model) simulations using COMSOL for validation. We find three evaporation regimes depending on the flow rate as well as on the distance of the gas outlet and the evaporating droplet. In one of the sample configurations tested the evaporation takes place in such a way that it can be described with the help of the simple diffusive model without convection. Thus, the presented approach opens comparative measurements with other methods as well as the observation of droplet evaporation in very dry or very humid environments with and without the influence of convection. Finally, using PRESS spectra, it is shown that the evaporation rate of water from a water/DMSO droplet can be controlled. This shows how the setup presented here can be used to study the evaporation of droplets of more complex mixtures.

Pseudogenes in ribonuclease evolution: a source of new biomacromolecular function?

Bovine seminal ribonuclease (RNase) diverged from pancreatic RNase after a gene duplication ca. 35 million years ago. Members of the seminal RNase gene family evidently remained as unexpressed pseudogene for much of its evolutionary history. Between 5 and 10 million years ago, however, after the divergence of kudu but before the divergence of ox, evidence suggests that the pseudogene was repaired and expressed. Intriguingly, detailed analysis of the sequences suggests that the repair may have involved gene conversion, transfer of information from the pancreatic gene to the RNase pseudogene. Further, the ratio of non-silent to silent substitutions suggests that the pancreatic RNases are divergently evolving under functional constraints, the seminal RNase pseudogenes are diverging under no functional constraints, while the genes expressed in the seminal plasma are evolving extremely rapidly in their amino acid sequences, as if to fulfil a new physiological role.

Developing new synthetic catalysts. How nature does it.

Paleomolecular biochemistry is a new field of science that seeks to understand how life emerged and developed in interaction with its geophysical surroundings. It is an experimental science, involving reconstruction of extinct biomolecules in the laboratory, studying their properties in the laboratory, and inferring details of their behavior and function in the context of geological data. An outline is provided of some tools of this field, together with its application to the study of two specific systems, ribonuclease and alcohol dehydrogenase.

Immunosuppressive activity of angiogenin in comparison with bovine seminal ribonuclease and pancreatic ribonuclease.

Angiogenin, a member of the pancreatic-like ribonuclease family with a special biological action (RISBAses), is a basic protein that induces blood vessel formation. Another member of these special ribonucleases, bovine seminal ribonuclease (BS RNase), displays biological properties, including aspermatogenic, embryotoxic, antitumor and immunosuppressive activities. The effects of two angiogenin preparations tested on the biological activities mentioned above are reported and compared with those of BS RNase and RNase A. In contrast to RNase A, which was ineffective in all biological activities tested, angiogenin suppressed significantly the proliferation of human lymphocytes stimulated by phytohemagglutinin or concanavalin A or by allogenic human lymphocytes (mixed lymphocyte culture). However, angiogenin did not affect the growth of human tumor cell lines, development of cow and mouse embryos and spermatogenicity in mice. On the basis of these results, angiogenin is the first monomeric ribonuclease described so far that displays immunosuppressive activity similar to that of the dimeric BS RNase. The immunosuppressive activity of angiogenin might synergize with the effect on neovascularization of tumor tissues and thus contribute to the development of tumor.

Site-specific cleavage of chromosomes in vitro through Cre-lox recombination.

Site-specific recombination systems are useful tools for chromosome engineering in vivo and site-specific DNA cleavage methods have applications in genome analysis and gene isolation. Here, we report a new method to fragment chromosomes in vitro using the Cre-lox site-specific recombination system. Two lox sites were targeted into the 5.7 Mb chromosomes I of Schizosaccharomyces pombe. In vitro recombination between chromosomal lox sites and exogenously provided lox oligonucleotides 'cleaved' the chromosome at the defined lox sequences. Site-specific cleavage of lox sites in the tobacco genome was also demonstrated. This recombination-based cleavage method provides a novel approach for structural and functional analyses of eukaryotic chromosomes as it allows direct isolation of chromosome regions that correspond to phenotypes revealed through Cre-lox mediated chromosome rearrangements in vivo. Moreover, recombination with end-labeled lox oligonucleotides would permit the specific end-labeling of chromosome segments to facilitate the long range mapping of chromosomes.

Cloning and expression of goldfish opsin sequences.

Five opsin cDNA clones were isolated from a goldfish retina cDNA library and sequenced. On the basis of homology to previously characterized visual pigments, one clone was identified as goldfish rod opsin and a second as a goldfish red cone opsin. Two rhodopsin-like clones were found to be similar to the chicken green opsin, a pigment which shares properties with both rod and cone pigments. A fifth clone was equally homologous to human blue cone opsin and human rod opsin. In order to characterize the spectral properties of the encoded pigments, the five clones were expressed in tissue culture cells and the apoproteins reconstituted with 11-cis-retinal. The wavelength of maximal absorption for goldfish rhodopsin is 492 nm and for the fifth pigment, identified as the goldfish blue pigment, 441 nm. Pigments encoded by the two rhodopsin-like clones absorb at 505 and 511 nm and are likely to correspond to the goldfish green pigment previously characterized by microspectrophotometry. The putative red cone opsin cDNA may encode a pigment that is a polymorphic variant of goldfish red since it absorbs maximally at 525 nm.