Uncertainties in the determination of pyrrolizidine alkaloid levels in naturally contaminated honeys and comparison of results obtained by different analytical approaches

The contamination of honey with hepatotoxic pyrrolizidine alkaloids (PAs) is a well-known hazard for food safety. While management strategies and controls of the honey industry aim to reduce the PA levels, uncertainties remain with regard to the safety of regionally produced and marketed honey. In addition, a previous study showed large differences of results obtained after various periods of storage and apparent differences between the analytical results of different laboratories. Therefore, this study aimed at examining these uncertainties by monitoring the impact of storage on the PA and PA N-oxide (PANO) content of two freshly harvested honeys and on possible demixing effects caused by pollen settling. Additionally, three analytical approaches – target analysis with matrix-matched calibration or standard addition and a sum parameter method – were applied for a comparative analysis of 20 honeys harvested in summer 2016. All samples originated from Schleswig-Holstein in Northern Germany where the PA plant Jacobaea vulgaris is currently observed on a massive scale. The results of the time series analyses showed that PANO levels markedly decreased within a few weeks and practically reached the LOD 16 weeks after harvest. Tertiary PAs, by contrast, remained stable and did not increase as a consequence of PANO decrease. The experiments on a putative demixing, which may result in a heterogeneous distribution of PAs/PANOs, revealed that there was no such effect during storage of up to 12 weeks. A comparison of the PA/PANO levels obtained by different analytical approaches showed that in some cases the sum parameter method yielded much higher levels than the target approaches, whereas in other cases, the target analysis with standard addition found higher levels than the other two methods. In summary, the results of this study highlight uncertainties regarding the validity and comparability of analytical results and consequently regarding health risk assessment.     

Pull-Down of Metalloproteins in Their Native States by Using Desthiobiotin-Based Probes

One-third of all proteins are estimated to require metals for structural stability and/or catalytic activity. Desthiobiotin probes containing metal binding groups can be used to capture metalloproteins with exposed active-site metals under mild conditions so as to prevent changes in metallation state. The proof-of-concept was demonstrated with carbonic anhydrase (CA), an open active site, Zn²⁺-containing protein. CA was targeted by using sulfonamide derivatives. Linkers of various lengths and structures were screened to determine the optimal structure for capture of the native protein. The optimized probes could selectively pull down CA from red blood cell lysate and other protein mixtures. Pull-down of differently metallated CAs was also investigated.     

Fabrication of a heterodyne receiver OEIC with optimized integration process using three MOVPE growth steps only

An optimized integration process is reported requiring only three epitaxial growth steps for the fabrication of an OEIC encompassing a single-mode laser, a waveguide element, and fast photodiodes. The process was applied to the fabrication of a heterodyne receiver including a semi-insulating buried heterostructure (SIBH)-DFB-laser with an output power of 14 mW and a side-mode suppression ratio greater than 40 dB, a 3-dB coupler, and high-speed balanced photodiodes with a bandwidth of 9 GHz. The laser can also be used as a transmitter laser exhibiting a modulation bandwidth of 11 GHz.     

Solubilization of fish muscle proteins with buffers containing sodium dodecyl sulfate

Summary The extraction of fish muscle protein using SDS containing solubilization buffers was studied varying the time and the temperature of solubilization, as well as pH and SDS concentration of the buffer. At pH < 6 the myofbrillar proteins were incompletely solubilized; temperatures of 80-100 °C resulted in protein degradation observable in the SDS-PAGE.Samples of fish muscle containing high amounts of formaldehyde (50 mmoles FA/kg wet weight) could only be solubilised at 100 °C; on the other hand it was possible to solubilize cooked and/or canned products under mild conditions (2% SDS, 1% 2-ME, pH 8.9, shaking for 2 h at 60 °C).

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Solubilisierung von Fischmuskelproteinen durch Natriumdodecylsulfathaltige Puffer

Zusammenfassung Der Einfluß von Solubilisierungszeit und -temperatur, des pH-Wertes und der SDS-Konzentration des Solubilisierungspuffers auf die Extraktion von Fischmuskelproteinen wurde überprüft. Bei pH-Werten < 6 wurden die myofbrillären Proteine nur unvollständig gelöst; Solubilisierungstemperaturen von 80 bis 100 °C führten zu einem in der SDS-PAGE sichtbaren Proteinabbau. Fischmuskelproben mit hohem Formaldehydgehalt (50 mmole FA/kg Feuchtgewicht) ließen sich nur bei 100 °C vollständig solubilisieren; demgegenüber gelang die weitgehende Solubilisierung gegarter und/oder sterilisierter Produkte unter milden Bedingungen (2% SDS, 1% 2-ME, pH 8,9; 2stündiges Schütteln bei 60 °C).

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Abbreviations FA formaldehyde - IEF isoelectric focusing - 2-ME 2-mercaptoethanol - PA polyacrylamide - PAGE polyacrylamide gel electrophoresis - pI isoelectric point - SDS sodium dodecyl sulfate - TMAO trimethylamine oxide - Tris Tris (hydroxymethyl)methylamine     

Determination of the formaldehyde content in fishery products

The influence of external factors, such as storage temperature and time on the content of free formaldehyde (FA) in fishery products is described. On the basis of the examination of several methods for the determination of free and bound FA, the following procedures are recommended: (1) for measuring free FA, the samples are extracted using 6% perchloric acid at room temperature and the FA content of the extracts is measured by the formation of 3,5-diacetyl-1,4-dihydrolutidine; (2) bound, acid-labile FA is released by steam distillation using 1% sulphuric acid, giving a pH value of about 1. The FA content of the distillates may be determined either by chromotropic acid assay or by the method described for the extracts.     

Identifying the criterion spontaneously minimized during the take-off phase of a sub-maximal long jump through optimal synthesis

Optimal synthesis of human movement or the prediction of the kinematics of a new movement require not only that the multi-body system be modeled but also that a performance criterion is specified. For sub-maximal movements the selection of a suitable performance criterion, able to generate realistic dynamic behavior is difficult. A two-dimensional simulation model of the take-off phase of a sub-maximal long jump was developed to study the effect of criterion choice on the realism of simulated movements. A parametric optimization technique was employed to obtain solutions to the constrained equations of motion. Seven different criteria were evaluated, by comparing simulated movements with an actual performance, to identify the criterion which most closely approximated that spontaneously minimized by the athlete. Synthesis of the take-off phase of a sub-maximal long jump was found to be sensitive to the chosen criterion, with a criterion based on minimizing joint intersegmental forces found to perform well.     

Nondestructive characterization of the mechanical strength of diffusion bonds. II. Application of a quasi-static spring model

It has been shown that the acoustic response of imperfect interfaces may be described by a quasi-static spring model. In the present paper, experimental data on the geometry of the contacts between two diffusion-bonded blocks have been used to determine the spring stiffness of such interfaces which have been correlated with experimentally determined ultrasonic reflection coefficients. The correlation between the theoretical reflection coefficient and the spring stiffness was found to be in excellent agreement with experimentally-observed values, if the disbonds are of infinitesimally small thickness. For disbonds of finite thickness, the agreement is less satisfactory. Reasons for the discrepancy in the latter case are unknown at the present time.     

Color object segmentation and tracking using flexible statistical model and level-set

Multimedia Tools and Applications - This study presents an unsupervised novel algorithm for color image segmentation, object detection and tracking based on unsupervised learning step followed with...     

Effect of substrate surface roughness on the adherence of NiCrAlY thermal spray coatings

The adherence of plasma sprayed NiCrAlY bond coats can be improved by an appropriate substrate surface finish. The interface fracture energy for crack propagation along the coating/substrate interface has been measured for different surface roughness by means of a specially designed four-point bending test. An increase of the interface fracture energy of about 15% was observed for a three times higher surface roughness. In addition, four-point bending tests with the coating on the side face of bending specimens were performed to analyze the fracture and spalling behavior of the coatings both under large tensile and compressive substrate deformations.