Lysosomal membrane stability in mussels

In 2012, the ICES Study Group on Integrated Monitoring of Chemicals and their Effects provided a framework for integrated monitoring to the OSLO-Paris Commission. UNEP/MAP and HELCOM expert groups have also developed guidelines on integrated monitoring of chemicals and their effects for the Mediterranean and Baltic Sea. This document provides the technical information for one of the biological effects measurements, the lysosomal membrane stability (LMS), which is a part of the above mentioned integrated monitoring approaches. Lysosomes are cytoplasmic, single membrane organelles whose condition is sensitive to stress whether it be due to environmental conditions or exposure to a wide array of contaminants. Two different methodologies have been developed to assess LMS in mussels: an enzyme cytochemical method using cryostatic sections of digestive gland tissue, and an in vivo cytochemical method (using haemolymph cells). In this document, different aspects of the operational procedures have been standardized and harmonized, with particular reference to the in vivo cytochemical method. New graphical material has been added to clarify criteria of interpretation and new external quality assurance programmes for measurements of lysosomal membrane stability have been proposed. Background (BAC) and environmental (EAC) assessment criteria to assess the LMS data are provided. Additionally, a new scoring procedure to enhance the sensitivity of the LMS measurements using the in vivo assay is provided.Versión del edito

Predicting the mechanism of phospholipidosis.

The mechanism of phospholipidosis is still not well understood. Numerous different mechanisms have been proposed, varying from direct inhibition of the breakdown of phospholipids to the binding of a drug compound to the phospholipid, preventing breakdown. We have used a probabilistic method, the Parzen-Rosenblatt Window approach, to build a model from the ChEMBL dataset which can predict from a compound's structure both its primary pharmaceutical target and other targets with which it forms off-target, usually weaker, interactions. Using a small dataset of 182 phospholipidosis-inducing and non-inducing compounds, we predict their off-target activity against targets which could relate to phospholipidosis as a side-effect of a drug. We link these targets to specific mechanisms of inducing this lysosomal build-up of phospholipids in cells. Thus, we show that the induction of phospholipidosis is likely to occur by separate mechanisms when triggered by different cationic amphiphilic drugs. We find that both inhibition of phospholipase activity and enhanced cholesterol biosynthesis are likely to be important mechanisms. Furthermore, we provide evidence suggesting four specific protein targets. Sphingomyelin phosphodiesterase, phospholipase A2 and lysosomal phospholipase A1 are shown to be likely targets for the induction of phospholipidosis by inhibition of phospholipase activity, while lanosterol synthase is predicted to be associated with phospholipidosis being induced by enhanced cholesterol biosynthesis. This analysis provides the impetus for further experimental tests of these hypotheses.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

A functional description of CymA, an electron-transfer hub supporting anaerobic respiratory flexibility in Shewanella

CymA (tetrahaem cytochrome c) is a member of the NapC/NirT family of quinol dehydrogenases. Essential for the anaerobic respiratory flexibility of shewanellae, CymA transfers electrons from menaquinol to various dedicated systems for the reduction of terminal electron acceptors including fumarate and insoluble minerals of Fe(III). Spectroscopic characterization of CymA from Shewanella oneidensis strain MR-1 identifies three low-spin His/His co-ordinated c-haems and a single high-spin c-haem with His/H2O co-ordination lying adjacent to the quinol-binding site. At pH 7, binding of the menaquinol analogue, 2-heptyl-4-hydroxyquinoline-N-oxide, does not alter the mid-point potentials of the high-spin (approximately −240 mV) and low-spin (approximately −110, −190 and −265 mV) haems that appear biased to transfer electrons from the high- to low-spin centres following quinol oxidation. CymA is reduced with menadiol (Em=−80 mV) in the presence of NADH (Em=−320 mV) and an NADH–menadione (2-methyl-1,4-naphthoquinone) oxidoreductase, but not by menadiol alone. In cytoplasmic membranes reduction of CymA may then require the thermodynamic driving force from NADH, formate or H2 oxidation as the redox poise of the menaquinol pool in isolation is insufficient. Spectroscopic studies suggest that CymA requires a non-haem co-factor for quinol oxidation and that the reduced enzyme forms a 1:1 complex with its redox partner Fcc3 (flavocytochrome c3 fumarate reductase). The implications for CymA supporting the respiratory flexibility of shewanellae are discussed.</jats:p

Neural Predictive Control of Broiler Chicken Growth

Active control of the growth of broiler chickens has potential benefits for farmers in terms of improved production efficiency, as well as for animal welfare in terms of improved leg health. In this work, a differential recurrent neural network (DRNN) was identified from experimental data to represent broiler chicken growth using a recently developed nonlinear system identification algorithm. The DRNN model was then used as the internal model for nonlinear model predicative control (NMPC) to achieve a group of desired growth curves. The experimental results demonstrated that the DRNN model captured the underlying dynamics of the broiler growth process reasonably well. The DRNN based NMPC was able to specify feed intakes in real time so that the broiler weights accurately followed the desired growth curves ranging from $-12$ to +12% of the standard curve. The overall mean relative error between the desired and achieved broiler weight was 1.8% for the period from day 12 to day 51

Black Hole Complementarity vs. Locality

The evaporation of a large mass black hole can be described throughout most of its lifetime by a low-energy effective theory defined on a suitably chosen set of smooth spacelike hypersurfaces. The conventional argument for information loss rests on the assumption that the effective theory is a local quantum field theory. We present evidence that this assumption fails in the context of string theory. The commutator of operators in light-front string theory, corresponding to certain low-energy observers on opposite sides of the event horizon, remains large even when these observers are spacelike separated by a macroscopic distance. This suggests that degrees of freedom inside a black hole should not be viewed as independent from those outside the event horizon. These nonlocal effects are only significant under extreme kinematic circumstances, such as in the high-redshift geometry of a black hole. Commutators of space-like separated operators corresponding to ordinary low-energy observers in Minkowski space are strongly suppressed in string theory.Comment: 32 pages, harvmac, 3 figure

Acute Inflammatory Profiles Differ with Sex and Age After Spinal Cord Injury

Background Sex and age are emerging as influential variables that affect spinal cord injury (SCI) recovery. Despite a changing demographic towards older age at the time of SCI, the effects of sex or age on inflammation remain to be elucidated. This study determined the sex- and age-dependency of the innate immune response acutely after SCI. Methods Male and female mice of ages 4- and 14-month-old received T9 contusion SCI and the proportion of microglia, monocyte-derived macrophages (MDM), and neutrophils surrounding the lesion were determined at 3- and 7-day post-injury (DPI) using flow cytometry. Cell counts of microglia and MDMs were obtained using immunohistochemistry to verify flow cytometry results at 3-DPI. Microglia and MDMs were separately isolated using fluorescence-activated cell sorting (FACS) at 3-day post-injury (DPI) to assess RNA expression of 27 genes associated with activation, redox, and debris metabolism/clearance. Results Flow cytometry revealed that being female and older at the time of injury significantly increased MDMs relative to other phagocytes, specifically increasing the ratio of MDMs to microglia at 3-DPI. Cell counts using immunohistochemistry revealed that male mice have more total microglia within SCI lesions that can account for a lower MDM/microglia ratio. With NanoString analyses of 27 genes, only 1 was differentially expressed between sexes in MDMs; specifically, complement protein C1qa was increased in males. No genes were affected by age in MDMs. Only 2 genes were differentially regulated in microglia between sexes after controlling for false discovery rate, specifically CYBB (NOX2) as a reactive oxygen species (ROS)-associated marker as well as MRC1 (CD206), a gene associated with reparative phenotypes. Both genes were increased in female microglia. No microglial genes were differentially regulated between ages. Differences between microglia and MDMs were found in 26 of 27 genes analyzed, all expressed higher in MDMs with three exceptions. Specifically, C1qa, cPLA2, and CD86 were expressed higher in microglia. Conclusions These findings indicate that inflammatory responses to SCI are sex-dependent at both the level of cellular recruitment and gene expression

Multi-nuclear, high-pressure, operando FlowNMR spectroscopic study of Rh/PPh3 – catalysed hydroformylation of 1-hexene

The hydroformylation of 1-hexene with 12 bar of 1 : 1 H2/CO in the presence of the catalytic system [Rh(acac)(CO)2]/PPh3 was successfully studied by real-time multinuclear high-resolution FlowNMR spectroscopy at 50 °C. Quantitative reaction progress curves that yield rates as well as chemo- and regioselectivities have been obtained with varying P/Rh loadings. Dissolved H2 can be monitored in solution to ensure true operando conditions without gas limitation. 31P{1H} and selective excitation 1H pulse sequences have been periodically interleaved with 1H FlowNMR measurements to detect Rh–phosphine intermediates during the catalysis. Stopped-flow experiments in combination with diffusion measurements and 2D heteronuclear correlation experiments showed the known tris-phosphine complex [RhH(CO)(PPh3)3] to generate rapidly exchanging isomers of the bis-phosphine complex [Rh(CO)2(PPh3)2] under CO pressure that directly enter the catalytic cycle. A new mono-phosphine acyl complex has been identified as an in-cycle reaction intermediate