Effects of lipid kinase expression and cellular stimuli on phosphatidylinositol 5-phosphate levels in mammalian cell lines

AbstractPhosphatidylinositol 5-phosphate (PtdIns5P) is a relatively recently discovered inositol lipid whose metabolism and functions are not yet clearly understood. We have transfected cells with a number of enzymes that are potentially implicated in the synthesis or metabolism of PtdIns5P, or subjected cells to a variety of stimuli, and then measured cellular PtdIns5P levels by a specific mass assay. Stable or transient overexpression of Type IIα PtdInsP kinase, or transient overexpression of Type Iα or IIβ PtdInsP kinases caused no significant change in cellular PtdIns5P levels. Similarly, subjecting cells to oxidative stress or EGF stimulation had no significant effect on PtdIns5P, but stimulation of HeLa cells with a phosphoinositide-specific PLC-coupled agonist, histamine, caused a 40% decrease within 1min. Our data question the degree to which inositide kinases regulate PtdIns5P levels in cells, and we discuss the possibility that a significant part of both the synthesis and removal of this lipid may be regulated by phosphatases and possibly phospholipases

In Vivo Biotransformation of 3,3′,4,4′-Tetrachlorobiphenyl by Whole Plants−Poplars and Switchgrass

Polychlorinated biphenyls (PCBs) are widely distributed persistent organic pollutants. In vitro research has shown that plant cell cultures might transform lower chlorinated congeners to hydroxylated PCBs, but there are few studies on in vivo metabolism of PCBs by intact whole plants. In this research, poplar plants (Populus deltoides × nigra, DN34) and switchgrass (Panicum vigratum, Alamo) were hydroponically exposed to 3,3′,4,4′-tetrachlorobiphenyl (CB77). Metabolism in plants occurred rapidly, and metabolites were detected after only a 24 h exposure. Rearrangement of chlorine atoms and dechlorination of CB77 by plants was unexpectedly observed. In addition, poplars were able to hydroxylate CB77 and the metabolite 6-hydroxy-3,3′,4,4′-tetrachlorobiphenyl (6-OH-CB77) was identified and quantified. Hybrid poplar was able to hydroxylate CB77, but switchgrass was not, suggesting that enzymatic transformations are plant specific. Sulfur-containing metabolites (from the action of sulfotransferases) were investigated in this study, but they were not detected in either poplar or switchgrass

Levels and enantiomeric signatures of methyl sulfonyl PCB and DDE metabolites in livers of harbor porpoises (Phocoena phocoena) from the southern North Sea

The concentration of 26 methyl sulfonyl metabolites of polychlorinated biphenyls (MeSO2-PCBs) and of p,p'-DDE (MeSO2-DDE) were determined in 19 liver samples from harbor porpoises (Phocoena phocoena) stranded between 1997 and 2000 on the Belgian and French North Sea Coasts. The total concentration of MeSO2-PCBs ranged from 39 to 4221 ng/g lipid weight (lw) and were generally higher in adults (age >2 yr, range 969-4221 ng/g lw) than in juveniles (age 0.73 or EF < 0.23) for the measured chiral MeSO2-PCB congeners was found in all samples. This result may suggest that one atropisomer may be preferentially formed in harbor porpoises or that the atropisomers are retained in a highly selective manner.Peer reviewe

Organophosphate esters (OPEs)in Chinese foodstuffs: Dietary intake estimation via a market basket method, and suspect screening using high-resolution mass spectrometry

Despite of the ubiquity of organophosphate esters (OPEs)in various environmental matrices, information regarding the dietary intakes of OPEs is currently limited. To better understand dietary exposure and intake, the present study investigated 11 OPE flame retardants (FRs)in 105 co

Closing the gap between science and management of cold-water refuges in rivers and streams

Human activities and climate change threaten coldwater organisms in freshwater eco-systems by causing rivers and streams to warm, increasing the intensity and frequency of warm temperature events, and reducing thermal heterogeneity. Cold-water refuges are discrete patches of relatively cool water that are used by coldwater organisms for thermal relief and short-term survival. Globally, cohesive management approaches are needed that consider interlinked physical, biological, and social factors of cold-water refuges. We review current understanding of cold-water refuges, identify gaps between science and management, and evaluate policies aimed at protecting thermally sensitive species. Existing policies include designating cold-water habitats, restricting fishing during warm periods, and implementing threshold temperature standards or guidelines. However, these policies are rare and uncoordinated across spatial scales and often do not consider input from Indigenous peoples. We propose that cold-water refuges be managed as dis-tinct operational landscape units, which provide a social and ecological context that is relevant at the watershed scale. These operational landscape units provide the founda-tion for an integrated framework that links science and management by (1) mapping and characterizing cold-water refuges to prioritize management and conservation actions, (2) leveraging existing and new policies, (3) improving coordination across jurisdictions, and (4) implementing adaptive management practices across scales. Our findings show that while there are many opportunities for scientific advancement, the current state of the sciences is sufficient to inform policy and management. Our proposed framework pro-vides a path forward for managing and protecting cold-water refuges using existing and new policies to protect coldwater organisms in the face of global change. behavioral thermoregulation, climate change adaptation, lotic ecosystem management, refugia, salmonids, temperature, thermal heterogeneity, thermal refugespublishedVersio

Finding needles in haystacks: linking scientific names, reference specimens and molecular data for Fungi

DNA phylogenetic comparisons have shown that morphology-based species recognition often underestimates fungal diversity. Therefore, the need for accurate DNA sequence data, tied to both correct taxonomic names and clearly annotated specimen data, has never been greater. Furthermore, the growing number of molecular ecology and microbiome projects using high-throughput sequencing require fast and effective methods for en masse species assignments. In this article, we focus on selecting and re-annotating a set of marker reference sequences that represent each currently accepted order of Fungi. The particular focus is on sequences from the internal transcribed spacer region in the nuclear ribosomal cistron, derived from type specimens and/or ex-type cultures. Re-annotated and verified sequences were deposited in a curated public database at the National Center for Biotechnology Information (NCBI), namely the RefSeq Targeted Loci (RTL) database, and will be visible during routine sequence similarity searches with NR_prefixed accession numbers. A set of standards and protocols is proposed to improve the data quality of new sequences, and we suggest how type and other reference sequences can be used to improve identification of Fungi