Adaption of a fragment analysis technique to an automated high-throughput multicapillary electrophoresis device for the precise qualitative and quantitative characterization of microbial communities

The analysis of microbial communities is of increasing importance in life sciences and bioengineering. Traditional techniques of investigations like culture or cloning methods suffer from many disadvantages. They are unable to give a complete qualitative and quantitative view of the total amount of microorganisms themselves, their interactions among each other and with their environment. Obviously, the determination of static or dynamic balances among microorganisms is of fast growing interest. The generation of species specific and fluorescently labeled 16S ribosomal DNA (rDNA) fragments by the terminal restriction fragment length polymorphism (T-RFLP) technique is a suitable tool to overcome the problems other methods have. For the separation of these fragments polyacrylamide gel sequencers are preferred as compared to capillary sequencers using linear polymers until now because of their higher electrophoretic resolution and therefore sizing accuracy. But modern capillary sequencers, especially multicapillary sequencers, offer an advanced grade of automation and an increased throughput necessary for the investigation of complex communities in long-time studies. Therefore, we adapted a T-RFLP technique to an automated high-throughput multicapillary electrophoresis device (ABI 3100 Genetic Analysis) with regard to a precise qualitative and quantitative characterization of microbial communities.     

Isotachophoretic zone formation of serum albumin in different free fluid electrophoresis instruments

The isotachophoretic behavior of a model protein, serum albumin, was examined (i) by computer simulation, (ii) by capillary isotachophoresis in HPE 100 and Tachophor 2127, (iii) by continuous flow isotachophoresis in Elphor VaP 22 and the BIO-STREAM Separator and (iv) by recycling isotachophoresis in an apparatus of our own design. Variations in monitored zone shapes can be explained by differences in engineering aspects and fluid stabilization principles of the instruments.     

Naturalness and Theoretical Constraints on the Higgs Boson Mass

Arbitrary regularization dependent parameters in Quantum Field Theory are usually fixed on symmetry or phenomenology grounds. We verify that the quadratically divergent behavior responsible for the lack of naturalness in the Standard Model (SM) is intrinsically arbitrary and regularization dependent. While quadratic divergences are welcome for instance in effective models of low energy QCD, they pose a problem in the SM treated as an effective theory in the Higgs sector. Being the very existence of quadratic divergences a matter of debate, a plausible scenario is to search for a symmetry requirement that could fix the arbitrary coefficient of the leading quadratic behavior to the Higgs boson mass to zero. We show that this is possible employing consistency of scale symmetry breaking by quantum corrections. Besides eliminating a fine-tuning problem and restoring validity of perturbation theory, this requirement allows to construct bounds for the Higgs boson mass in terms of $\delta m^{2}/m^{2}_{H}$ (where m _H is the renormalized Higgs mass and δm ² is the 1-loop Higgs mass correction). Whereas $\delta m^{2}/m^{2}_{H}<1$ (perturbative regime) in this scenario allows the Higgs boson mass around the current accepted value, the inclusion of the quadratic divergence demands $\delta m^{2}/m^{2}_{H}$ arbitrarily large to reach that experimental value.     

Real-time increase in depth of field of an uncooled thermal camera using several phase-mask technologies

Imaging systems that combine a phase mask in the pupil and digital postprocessing may have better performance than conventional ones. We have built such a system to enhance the depth of field of an uncooled thermal camera. The phase masks are binary, their structures are optimized thanks to an image quality criterion, and they have been realized with three different technologies that give equivalent results. The deconvolution postprocessing is performed in real time with a graphics processing unit. A significant increase of the depth of field of a factor 3 has been obtained.     

Grain Boundary Grooving as an Indicator of Grain Boundary Phase Transformations

The atomic force microscopy (AFM) was used to study the grain boundary (GB) groove profiles far away from the melting temperature T _m. It is shown that AFM allows one to measure the temperature dependence of the GB energy in a rather broad temperature interval (from 0.85 T _m to T _m). The GB energy and GB segregation of Bi were measured at 1123 K in the interval of the Bi bulk concentration x ^v _Bi from 5 to 140 ppm Bi. The transition from monolayer to multilayer adsorption is observed for the 19a GB at 1123 K and x ^v _Bi = 60 at. ppm Bi. At the same point (1123 K and x ^v _Bi = 60 at. ppm Bi) a discontinuity of the first derivative of the GB energy is observed. These features were explained using the model of GB prewetting phase transformation developed previously.     

Intra- and intergroup vocal behavior in resident killer whales, Orcinus orca

Vocal communication within and between groups of individuals has been described extensively in birds and terrestrial mammals, however, little is known about how cetaceans utilize their sounds in their natural environment. Resident killer whales, Orcinus orca, live in highly stable matrilines and exhibit group-specific vocal dialects. Single call types cannot exclusively be associated with particular behaviors and calls are thought to function in group identification and intragroup communication. In the present study call usage of three closely related matrilines of the Northern resident community was compared in various intra- and intergroup contexts. In two out of the three matrilines significant changes in vocal behavior depending both on the presence and identity of accompanying whales were found. Most evidently, family-specific call subtypes, as well as aberrant and variable calls, were emitted at higher rates, whereas "low arousal" call types were used less in the presence of matrilines from different pods, subclans, or clans. Ways in which the observed changes may function both in intra- and intergroup communication.     

Investigation of NH–π (Ph) interaction in the 1:1[α:α-N-amino]mers using 1H NMR and FTIR absorption spectroscopy

¹H NMR and FTIR experiments performed on the 1:1[α:α-N-amino]mers series let us to point out a rare NH–π interaction between the carbamidic NH of the i residue and the phenyl group of a phenylalanine residue located at i+2.     

Effectiveness of natural and synthetic blocking reagents and their application for detecting food allergens in enzyme-linked immunosorbent assays

Blocking is an important step before an enzyme-linked immunosorbent assay (ELISA) can be performed. It reduces non-specific binding to the microtiter plate to a minimum. For detecting food allergens by means of ELISA, the problem with protein blocking solutions is obvious. The blocker might interfere with the antibodies of the assay and leads to false positive results. Therefore, other blocking solutions are greatly needed. There are some alternatives like synthetic blockers or carbohydrates. Comparisons of these different blocking agents, namely proteins, carbohydrates, and synthetic blockers, were made at different reaction conditions. The incubation periods and temperatures were varied, as well as the pH. The best combinations were evaluated and compared, in respect of their blocking efficiency. The two best non-proteinaceous blockers, i.e. polyvinylalcohol and Ficoll, were subsequently applied to ELISA tests for the determination of α-casein and peanut. The study showed that Ficoll and PVA did as well as BSA in buffer solution. Therefore, they can be considered as alternative blocking reagents for ELISA, especially for the detection of food allergens.     

Conformational properties of azapeptides

The conformation of several model compounds for azapeptides was systematically examined on the basis of ab initio MO theory at various approximation levels. The calculations show the azapeptide conformation essentially determined by the hydrazine and urea constituents along the sequence. This leads to a characteristic conformer pattern which excludes the possibility of β sheet conformations, but indicates a high potency for the formation of helix and β turn structures. The N atom may change between planar and pyramid structures. The peculiar conformation properties make azaamino acids an attractive tool for secondary structure design in peptides and proteins.     

Synthesis of 2,3-anti-3,4-anti and 2,3-anti-3,4-syn propionate motifs: a diastereoselective tandem sequence of Mukaiyama and free-radical-based hydrogen transfer reactions

[reaction: see text] Reported herein is a strategy employing a Mukaiyama reaction in tandem with a hydrogen transfer reaction for the elaboration of 2,3-anti-3,4-anti and 2,3-anti-3,4-syn propionate motifs. The mode of complexation is controlled through monodentate or chelate pathways for the Mukaiyama reaction to give access to either syn or anti aldol products, precursors of the free-radical reduction reaction. Boron Lewis acid is used to control the free-radical reaction through the exocyclic pathway.