Active Transport of Hepatotoxic Pyrrolizidine Alkaloids in HepaRG Cells

1,2-unsaturated pyrrolizidine alkaloids (PAs) are secondary plant metabolites occurring as food contaminants that can cause severe liver damage upon metabolic activation in hepatocytes. However, it is yet unknown how these contaminants enter the cells. The role of hepatic transporters is only at the beginning of being recognized as a key determinant of PA toxicity. Therefore, this study concentrated on assessing the general mode of action of PA transport in the human hepatoma cell line HepaRG using seven structurally different PAs. Furthermore, several hepatic uptake and efflux transporters were targeted with pharmacological inhibitors to identify their role in the uptake of the PAs retrorsine and senecionine and in the disposition of their N-oxides (PANO). For this purpose, PA and PANO content was measured in the supernatant using LC-MS/MS. Also, PA-mediated cytotoxicity was analyzed after transport inhibition. It was found that PAs are taken up into HepaRG cells in a predominantly active and structure-dependent manner. This pattern correlates with other experimental endpoints such as cytotoxicity. Pharmacological inhibition of the influx transporters Na⁺/taurocholate co-transporting polypeptide (SLC10A1) and organic cation transporter 1 (SLC22A1) led to a reduced uptake of retrorsine and senecionine into HepaRG cells, emphasizing the relevance of these transporters for PA toxicokinetics.     

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.     

Adventitial Tertiary Lymphoid Organs as Potential Source of MicroRNA Biomarkers for Abdominal Aortic Aneurysm

Abdominal aortic aneurysm (AAA) is an inflammatory disease associated with marked changes in the cellular composition of the aortic wall. This study aims to identify microRNA (miRNA) expression in aneurysmal inflammatory cells isolated by laser microdissection from human tissue samples. The distribution of inflammatory cells (neutrophils, B and T lymphocytes, mast cells) was evaluated in human AAA biopsies. We observed in half of the samples that adventitial tertiary lymphoid organs (ATLOs) with a thickness from 0.5 to 2 mm were located exclusively in the adventitia. Out of the 850 miRNA that were screened by microarray in isolated ATLOs (n = 2), 164 miRNAs were detected in ATLOs. The three miRNAs (miR-15a-3p, miR-30a-5p and miR-489-3p) with the highest expression levels were chosen and their expression quantified by RT-PCR in isolated ATLOs (n = 4), M1 (n = 2) and M2 macrophages (n = 2) and entire aneurysmal biopsies (n = 3). Except for the miR-30a-5p, a similar modulation was found in ATLOs and the two subtypes of macrophages. The modulated miRNAs were then evaluated in the plasma of AAA patients for their potential as AAA biomarkers. Our data emphasize the potential of miR-15a-3p and miR-30a-5p as biomarkers of AAA but also as triggers of ATLO evolution. Further investigations will be required to evaluate their targets in order to better understand AAA pathophysiology.     

Engineering egress data considering pedestrians with reduced mobility

To quantify the evacuation process, evacuation practitioners use engineering egress data describing the occupant movement characteristics. These data are typically based to young and fit populations. However, the movement abilities of occupants who might be involved in evacuations are becoming more variable—with the building populations of today typically including increasing numbers of individuals: with impairments or who are otherwise elderly or generally less mobile. Thus, there will be an increasing proportion of building occupants with reduced ability to egress. For safe evacuation, there is therefore a need to provide valid engineering egress data considering pedestrians with disabilities. Gwynne and Boyce recently compiled a series of data sets related to the evacuation process to support practitioner activities in the chapter Engineering Data in the SFPE Handbook of Fire Protection Engineering. This paper supplements these data sets by providing information on and presenting data obtained from additional research related to the premovement and horizontal movement of participants with physical‐, cognitive‐, or age‐related disabilities. The aim is to provide an overview of currently available data sets related to, and key factors affecting the egress performance of, mixed ability populations which could be used to guide fire safety engineering decisions in the context of building design.     

Reexamination of the Ergothioneine Biosynthetic Methyltransferase EgtD from Mycobacterium tuberculosis as a Protein Kinase Substrate

Ergothioneine has emerged as a crucial cytoprotectant in the pathogenic lifestyle of Mycobacterium tuberculosis. Production of this antioxidant from primary metabolites may be regulated by phosphorylation of Thr213 in the active site of the methyltransferase EgtD. The structure of mycobacterial EgtD suggests that this post-translational modification would require a large-scale change in conformation to make the active-site residue accessible to a protein kinase. In this report, we show that, under in vitro conditions, EgtD is not a substrate of protein kinase PknD.     

Bibliometric analysis of the evolution of biochar research trends and scientific production

Clean Technologies and Environmental Policy - Biochar has caught great attention over the last decade, yielding a large number of publications in a broad range of disciplines. This scientometric...     

Performance of PKI-based security mechanisms in mobile ad hoc networks

Security for ad hoc network environments has received a lot of attention as of today. Previous work has mainly been focussing on secure routing, fairness issues, and malicious node detection. However, the issue of introducing and conserving trust relationships has received considerably less attention. In this article, we present a scalable method for the use of public key certificates and their revocation in mobile ad hoc networks (MANETs). With the LKN-ad hoc security framework (LKN-ASF) a certificate management protocol has been introduced, bringing PKI technology to MANETs. In addition a performance analysis of two different revocation approaches for MANETs will be presented.     

Multibody simulation tool for the calculation of lashing loads on RoRo ships

Approximately 80% of the international transport of goods is carried on by means of ships. A large portion of the transport capacity is represented by Roll-on-Roll-off (RoRo) ships. Especially in Europe this is a relevant potential for the RoRo segment. Consequently, the design and construction of RoRo ships plays an increasing role for German shipyards and their suppliers. In order to make the loading and unloading procedure of trailer economically more competitive, ship owners would like to improve the lashing of trailers on the ship. On the basis of a multibody system formalism, a software tool has been developed which allows for an optimization of the loading of trailers on RoRo ships. Commemorative Contribution.     

Does {charm} need depth? Similarity and levels-of-processing effects in cued recall.

Eich (1985) recently presented a distributed memory model in which the pattern of results used to support the levels-of-processing view of Craik and Lockhart (1972) was modeled by different degrees of similarity between the encoding context and the to-be-recalled item. We report two experiments in which both phonemic and semantic similarity were varied between pairs of words and incidental acquisition (rhyme vs. category judgments) was varied across the same pairs of items. In both experiments the manipulation of the acquisition task produced a difference in cued-recall performance for positive and negative rhyme and category judgments. Recall was better following a category encoding decision than following a rhyme decision. This difference was independent of the effects of similarity, which demonstrated that Eich's (1985) assumptions regarding the effects of similarity are not sufficient to account for the differences resulting from the manner in which subjects encode information. An alternative method of modeling the levels-of-processing effect within the framework of distributed memory models is proposed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)