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.     

The Determinants of Debt and (Private) Equity Financing: The Case of Young, Innovative SMEs from Germany

Financial theory creates a puzzle. Some authors argue that high-risk entrepreneurs choose debt contracts instead of equity contracts since risky but high returns are of relatively more value for a loan-financed firm. Conversely, authors who focus explicitly on start-up finance predict that entrepreneurs are the more likely to seek equity-like venture capital contracts, the more risky their projects are. Our paper is an initial step towards resolving this puzzle empirically. We present microeconometric evidence on the determinants of debt and equity financing in young and innovative SMEs. We pay special attention to the role of risk for the choice of the method of financing. Since risk is not directly observable we use different indicators for financial and project risk. It turns out that our data generally confirms the hypothesis that the probability that a young high-tech firm receives equity financing is an increasing function of the financial risk. With regard to the intrinsic project risk, our results are less conclusive, as some of our indicators of a risky project are found to have a negative effect on the likelihood to be financed by private equity.     

Diurnal and spatial variation of xylem dielectric constant in Norway Spruce (Picea abies [L.] Karst.) as related to microclimate, xylem sap flow, and xylem chemistry

Spatial and temporal variations in vegetation dielectric properties strongly influence the microwave backscatter characteristics of forested landscapes. This paper examines the relationship between xylem tissue dielectric constant, xylem sap flux density, and xylem sap chemical composition as measured in the stems of two Norway Spruce (Picea abies [L.] Karst.) trees in the Fichtelgebirge region of Northern Bavaria, Germany. Dielectric constant and xylem sap flux were monitored continuously from June through October 1995, at several heights along the tree trunks. At the end of the measurement series, each tree was harvested, and its xylem sap extracted and analyzed to determine the concentrations of amino acids and cations. Results show that the sap flux density was correlated with vapor pressure deficit (VPD) at all heights in the stem. In contrast, the xylem tissue dielectric constant is influenced by VPD but can exhibit a significant temporal lag relative to changes in VPD. This lag varies with position along the tree trunk. The temporal variability of the dielectric constant is compared with both trees at several positions along the tree trunks. Results of xylem sap chemical analysis are presented. We show that spatial and temporal variability in the xylem tissue dielectric constant is influenced not only by water content, but by variations in xylem sap chemistry as well. This has important implications for microwave remote sensing of forested landscapes, as useful information may be acquired regarding stand physiology and water relations and where variations in dielectric properties within individual trees and across geographic areas can be significant error sources for forest inventory mapping.     

Structure-permeation relations of met-enkephalin peptide analogues on absorption and secretion mechanisms in Caco-2 monolayers

Due to the low effective permeabilities of peptides at many absorption sites, their structure-permeation relations are of high interest. In this work structure-permeation relations of Met-enkephalin analogues are presented using confluent Caco-2 cells as an in vitro permeation model. Four model peptides (Met-enkephalin, [D-Ala2]Met-enkephalin, [D-Ala2]Met-enkephalinamide, and metkephamid) were tested in terms of permeability, lipophilicity, charge, and molecular size. Permeability coefficients (P(eff)) across Caco-2 cells were low, 3.3 x 10(-8) to 9.5 x 10(-8) cm s-1, and were similar to typical paracellular markers. No correlation of permeability and the log(apparent octanol/buffer partition coefficient) was observed. A 40-fold increase of the permeability of metkephamid in the presence of 10 mM EDTA suggested a significant contribution of paracellular transport. Independent support for this conclusion was obtained by visualizing the pathway of the fluorescein isocyanate isomer I 1-metkephamid by confocal laser scanning microscopy (CLSM). The fluorophore-labeled peptide was observed in the intercallular space only. Metkephamid permeabilities were found to be direction-specific. Permeabilities from basolateral to apical (b-to-a) were significantly higher (ca. 4-fold) than in the opposite (a-to-b) direction. The addition of verapamil equalized the permeabilities in the a-to-b and b-to-a directions, suggesting the involvement of a P-glycoprotein-mediated secretion mechanism. Similar observations were obtained with [D-Ala2]Met-enkephalinamide, but not with Met-enkephalin and [D-Ala2]Met-enkephalin. In contrast to the other analogues, metkephamid and [D-Ala2]Met-enkephalinamide are positively charged at neutral pH, as demonstrated by their isoelectric points (pl = 8.6 for [D-Ala2]Met-enkephalinamide and metkephamid and 5.3 for [D-Ala2]Met-enkephalin and Met-enkephalin). The data is in agreement with the literature showing that most compounds secreted by the P-glycoprotein transporter carry a positive charge.     

A vectorial vibrating reed magnetometer with high sensitivity

A vibrating reed magnetometer with high sensitivity is constructed. Using optical signal pickup, modified excitation coils, and an acoustical reference excitation system, the high sensitivity of former constructions is combined with the detection of the magnetization component perpendicular to the bias field H. The calibration procedure providing increased reliability is simplified and allows hysteresis measurements with the magnetic easy axis of the sample oriented at an arbitrary angle with respect to H. Measurement results are given in order to demonstrate the performance of the apparatus     

Bipolar electric fatigue behaviour as a function of field strength in antiferroelectric (Pb,Ba,La)(Zr,Sn,Ti)O3 ceramics

The bipolar electric fatigue behaviour of antiferroelectric ceramics with the composition of Pb_0.88Ba_0.10La_0.02(Zr_0.55Sn_0.35Ti_0.10)O₃ was investigated under various cycling fields. The material exhibits a degradation in the maximum field induced strain, a diffuse AFE-FE phase transition and an enhancement in the diffusion character of the FE-AFE phase transition due to electric cycling. Those variations increase with cycle number, indicating a logarithmic fatigue up to 10⁸ cycles. There is no indication for the variations to be recovered, and the symmetry of the negative and positive parts of the strain hysteresis loops still remains. Higher cycling field results in a stronger deterioration of the maximum field induced strain and a larger extent of diffusion in AFE-FE and FE-AFE phase transitions. The normalized maximum strain shows nearly the same scale of degradation when the materials are cycled at various electric fields. After a heat treatment at 500°C for 1 h, the variations in the AFE-FE and FE-AFE phase transition due to cycling disappear, whereas the maximum strain resumes almost to its original value. Electrochemical variations are considered to contribute to the main fatigue mechanism for the material under investigation.