Biodegradability of 14C-labeled antibiotics in a modified laboratory scale sewage treatment plant at environmentally relevant concentrations

For all new pharmaceuticals, an environmental risk assessment (ERA) has to be performed according to guidelines developed by the European Medicines Evaluation Agency. An important factor of this procedure is the assessment of the predicted environmental concentration in the aquatic environment, which is significantly influenced by the biodegradability of pharmaceuticals in sewage treatment plants. Established standardized methods for determining biodegradation under laboratory conditions apply to substance concentrations, which are much higherthan those expected in reality. Against this background, the laboratory scale sewage treatment plant (LSSTP), as described by the Organisation for Economic Cooperation and Development (OECD) Guideline No. 303A, was modified to construct a lossless system, which allows laboratory testing at realistic concentrations. To verify the experimental setup, the antibiotics benzylpenicillin, ceftriaxone, and trimethoprim were tested at low concentrations (microg/L) using 14C-labeled compounds. The results show that approximately 25% of benzylpenicillin was mineralized, whereas ceftriaxone and trimethoprim were not mineralized at all. Due to the high total recoveries of added radioactivity (> or =95%) and the fact that the findings comply with available literature data, the lossless operation of the test system could be proved. Consequently, the modified LSSTP is a suitable tool to determine more realistic biodegradation data required for the exposure assessment within the scope of an ERA for pharmaceuticals.     

Calibration for cylindrical specimens in grazing-incidence interferometry via integration of difference measurements

Cylindrical specimens may be tested advantageously by using grazing-incidence interferometry. A multiple positions test in combination with rotational averaging has recently been used to separate the surface deviations of the specimen from the interferometric aberrations. To reduce the measuring time and to check whether the results are reliable, a second procedure is now investigated, which uses the principle of the multiple positions test to determine quantities proportional to the difference quotients of the surface deviations. After numerical integration, the results can be compared with those obtained previously by rotational averaging. The measurement principle is described, and calibration results are presented.     

Chromatic confocal spectral interferometry

Chromatic confocal spectral interferometry (CCSI) is a novel scheme for topography measurements that combines the techniques of spectral interferometry and chromatic confocal microscopy. This hybrid method allows for white-light interferometric detection with a high NA in a single-shot manner. To the best of our knowledge, CCSI is the first interferometric method that utilizes a confocally filtered and chromatically dispersed focus for detection and simultaneously allows for retrieval of the depth position of reflecting or scattering objects utilizing the phase (modulation frequency) of the interferometric signals acquired. With the chromatically dispersed focus, the depth range of the sensor is decoupled from the NA of the microscope objective.     

From brittle to ductile fracture of bone

Toughness is crucial to the structural function of bone. Usually, the toughness of a material is not just determined by its composition, but by the ability of its microstructure to dissipate deformation energy without propagation of the crack. Polymers are often able to dissipate energy by viscoplastic flow or the formation of non-connected microcracks. In ceramics, well-known toughening mechanisms are based on crack ligament bridging and crack deflection. Interestingly, all these phenomena were identified in bone, which is a composite of a fibrous polymer (collagen) and ceramic nanoparticles (carbonated hydroxyapatite). Here, we use controlled crack-extension experiments to explain the influence of fibre orientation on steering the various toughening mechanisms. We find that the fracture energy changes by two orders of magnitude depending on the collagen orientation, and the angle between collagen and crack propagation direction is decisive in switching between different toughening mechanisms.     

Study of the hydrophobic cavity of beta-cryptogein through laser-polarized xenon NMR spectroscopy

The interaction of xenon with beta-cryptogein, a basic 10 kDa protein belonging to the elicitin family, has been studied by using dissolved thermal and laser-polarized gas in liquid-state NMR. 13C and 1H chemical-shift-mapping experiments were unfruitful, the proton lines only experienced a slight narrowing but no significant frequency variation when the xenon concentration was increased. Nevertheless magnetization transfer from hyperpolarized xenon to protons of the protein demonstrates an undoubted interaction and enables localization of the noble-gas-binding site. Due to the proton-proton cross-relaxation efficiency, however, this experiment is subjected to important spin-diffusion. An automatic procedure that takes spin-diffusion into account when assigning the protons that interact with xenon is then used. The binding site, as defined by 30 Xe--H interactions, is situated in the inner core of the protein. The protons that interact with xenon border the channel by which sterols are known to enter into the cavity. These results support the idea that xenon is a good probe for hydrophobic protein regions.     

Modelling the structure of latexin-carboxypeptidase A complex based on chemical cross-linking and molecular docking

We have determined the three-dimensional structure of the protein complex between latexin and carboxypeptidase A using a combination of chemical cross-linking, mass spectrometry and molecular docking. The locations of three intermolecular cross-links were identified using mass spectrometry and these constraints were used in combination with a speed-optimised docking algorithm allowing us to evaluate more than 3 x 10(11) possible conformations. While cross-links represent only limited structural constraints, the combination of only three experimental cross-links with very basic molecular docking was sufficient to determine the complex structure. The crystal structure of the complex between latexin and carboxypeptidase A4 determined recently allowed us to assess the success of this structure determination approach. Our structure was shown to be within 4 A r.m.s. deviation of Calpha atoms of the crystal structure. The study demonstrates that cross-linking in combination with mass spectrometry can lead to efficient and accurate structural modelling of protein complexes.     

Reactions of chlorogenic acid and quercetin with a soy protein isolate--influence on the in vivo food protein quality in rats

Plant phenolic compounds are known to interact with proteins producing changes in the food (e. g., biological value (BV), color, taste). Therefore, the in vivo relevance, especially, of covalent phenol-protein reactions on protein quality was studied in a rat bioassay. The rats were fed protein derivatives at a 10% protein level. Soy proteins were derivatized with chlorogenic acid and quercetin (derivatization levels: 0.056 and 0.28 mmol phenolic compound/gram protein). Analysis of nitrogen in diets, urine, and fecal samples as well as the distribution of amino acids were determined. Depending on the degree of derivatization, the rats fed with soy-protein derivatives showed an increased excretion of fecal and urinary nitrogen. As a result, true nitrogen digestibility, BV, and net protein utilization were adversely affected. Protein digestibility corrected amino acid score was decreased for lysine, tryptophan, and sulfur containing amino acids.     

Estimation of measurement uncertainties using virtual fringe projection technique

One of the main tasks of the quality test is the inspection of all relevant geometric parts related to the predefined tolerance range, whereas the uncertainty of measurement has to be less than the tolerance range. The reachable uncertainty of measurement can be determined using method A of the ISO Guide to the Expression of Uncertainty in Measurement (GUM), which is expensive and time consuming and has to be carried out for each individual metrologic case. Furthermore, it is possible to check the suitability of the measurement system for the planned inspection using virtual measurement techniques and therewith to reduce the time and money spent. This means that the uncertainty of measurement is estimated using method B of the GUM. In this paper, a virtual fringe projection system is used for the estimation of the uncertainty of measurement, which is compared with the uncertainty of measurement determined with a real measurement system using method A of the GUM. With the presented method, it is possible to calculate an optimal measurement position within the measurement volume, based on a minimum uncertainty of measurement. Thereby, the influence of the operator related to the uncertainty can be significantly reduced.     

Behavior of scaled-up sodium alanate hydrogen storage tanks during sorption

Sodium alanate is being experimentally tested in scaled-up quantities. For this purpose, several tanks have been designed and constructed. The tank functionality during absorption and desorption of hydrogen was demonstrated in a scale of 8 kg of alanate, with a peak technical absorption time below 10 min. The absorption and desorption data show good reproducibility. Neutron radiography was used in another tank to show the powder’s physical behavior during sorption, showing conservation of the macroscopic structure during cycling.