Imaging Mitochondrial Calcium Fluxes with Fluorescent Probes and Single- or Two-Photon Confocal Microscopy

The concentration of calcium ions in the mitochondria has been shown to affect its function, modulating respiratory activity at low levels and causing lethal damage at high concentrations. The rhodamine series of dyes can be used to measure mitochondrial calcium concentration, but the reliability of measurements depends upon correct partitioning of the dye within to the mitochondria. Methods are described to aid verification and quantification of the mitochondrial calcium concentration using single- or two-photon confocal microscopy. The method of linear unmixing to separate fluorescent signals based on either differing excitation or emission spectra is outlined and for the purposes of illustration is applied to the separation of rhod-2 signals originating from the dye within the mitochondria and nucleoli

Growth disturbance of extracts from several crops straw (residue) on Ageratina adenophora and biological-control implications in hazardous weed invasion for eco-restoration

Laboratory biological simulation experiment was conducted to investigate growth disturbance of high, moderate, low concentration of aqueous extracts (i.e. the original extracts with a solid liquid ratio of 1:40 g mL-1 and its 5 times diluents and 25 times diluents) from several crops straw (residue) on Ageratina adenophora, a worldwide notorious invasive weed. The results showed: (a) aqueous extracts from several crops straw (residue) brought about different impacts on the single index for germination and growth of A. adenophora, e.g., high concentration of aqueous extracts from Brassica oleracea waste leaves showed a strong inhibition against the germination rate (GR) and germination index (GI) of A. adenophora, while high concentration of aqueous extracts from Vicia cracca straw showed a strong inhibition against radicle length (RL) and hypocotyl length (HL) of A. adenophora; (b) high concentration of aqueous extracts from B. oleracea waste leaves and high, moderate and low concentration of aqueous extracts from Oryza sativa straw and Triticum aestivum straw showed rather strong synthetic effects (inhibition) on GR and GI of A. adenophora, which could be chosen for the control over the seeds germination of A. adenophora; (c) high and moderate concentrations of aqueous extracts from V. cracca straw, high concentration of aqueous extracts from B. campestris waste leaves, and moderate and low concentrations of aqueous extracts from O. sativa straw and T. aestivum straw showed rather strong synthetic effects (inhibition) on RL and HL of A. adenophora, which could be selected as ideal materials for the control over the seedlings growth of A. adenophora; and (d) high concentrations of aqueous extracts from V. cracca straw, B. oleracea waste leaves and B. campestris waste leaves, and high, moderate and low concentrations of aqueous extracts from O. sativa straw and T. aestivum straw showed rather strong synthetic effects (inhibition) on GR, GI, RL and HL of A. adenophora, which could be selected as ideal materials for the control over the seeds germination and seedlings growth of A. adenophora. Thus, this study would provide a theoretic guidance and technical support for the resources utilization of crops straw (residue) and the prevention and control over invasive weeds as well. (C) 2013 Elsevier B.V. All rights reserved.Laboratory biological simulation experiment was conducted to investigate growth disturbance of high, moderate, low concentration of aqueous extracts (i.e. the original extracts with a solid liquid ratio of 1:40 g mL-1 and its 5 times diluents and 25 times diluents) from several crops straw (residue) on Ageratina adenophora, a worldwide notorious invasive weed. The results showed: (a) aqueous extracts from several crops straw (residue) brought about different impacts on the single index for germination and growth of A. adenophora, e.g., high concentration of aqueous extracts from Brassica oleracea waste leaves showed a strong inhibition against the germination rate (GR) and germination index (GI) of A. adenophora, while high concentration of aqueous extracts from Vicia cracca straw showed a strong inhibition against radicle length (RL) and hypocotyl length (HL) of A. adenophora; (b) high concentration of aqueous extracts from B. oleracea waste leaves and high, moderate and low concentration of aqueous extracts from Oryza sativa straw and Triticum aestivum straw showed rather strong synthetic effects (inhibition) on GR and GI of A. adenophora, which could be chosen for the control over the seeds germination of A. adenophora; (c) high and moderate concentrations of aqueous extracts from V. cracca straw, high concentration of aqueous extracts from B. campestris waste leaves, and moderate and low concentrations of aqueous extracts from O. sativa straw and T. aestivum straw showed rather strong synthetic effects (inhibition) on RL and HL of A. adenophora, which could be selected as ideal materials for the control over the seedlings growth of A. adenophora; and (d) high concentrations of aqueous extracts from V. cracca straw, B. oleracea waste leaves and B. campestris waste leaves, and high, moderate and low concentrations of aqueous extracts from O. sativa straw and T. aestivum straw showed rather strong synthetic effects (inhibition) on GR, GI, RL and HL of A. adenophora, which could be selected as ideal materials for the control over the seeds germination and seedlings growth of A. adenophora. Thus, this study would provide a theoretic guidance and technical support for the resources utilization of crops straw (residue) and the prevention and control over invasive weeds as well. (C) 2013 Elsevier B.V. All rights reserved

Environmental drivers of distribution and reef development of the Mediterranean coral Cladocora caespitosa

Cladocora caespitosa is the only Mediterranean scleractinian similar to tropical reef-building corals. While this species is part of the recent fossil history of the Mediterranean Sea, it is currently considered endangered due to its decline during the last decades. Environmental factors affecting the distribution and persistence of extensive bank reefs of this endemic species across its whole geographic range are poorly understood. In this study, we examined the environmental response of C. caespitosa and its main types of assemblages using ecological niche modeling and ordination analysis. We also predicted other suitable areas for the occurrence of the species and assessed the conservation effectiveness of Mediterranean marine protected areas (MPAs) for this coral. We found that phosphate concentration and wave height were factors affecting both the occurrence of this versatile species and the distribution of its extensive bioconstructions in the Mediterranean Sea. A set of factors (diffuse attenuation coefficient, calcite and nitrate concentrations, mean wave height, sea surface temperature, and shape of the coast) likely act as environmental barriers preventing the species from expansion to the Atlantic Ocean and the Black Sea. Uncertainties in our large-scale statistical results and departures from previous physiological and ecological studies are also discussed under an integrative perspective. This study reveals that Mediterranean MPAs encompass eight of the ten banks and 16 of the 21 beds of C. caespitosa. Preservation of water clarity by avoiding phosphate discharges may improve the protection of this emblematic species.Spanish Ministry of Economy and Competitiveness [CTM2014-57949-R]info:eu-repo/semantics/publishedVersio

Biochemical warfare on the reef : the role of glutathione transferases in consumer tolerance of dietary prostaglandins

© 2010 The Authors. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS ONE 5 (2010): e8537, doi:10.1371/journal.pone.0008537.Despite the profound variation among marine consumers in tolerance for allelochemically-rich foods, few studies have examined the biochemical adaptations underlying diet choice. Here we examine the role of glutathione S-transferases (GSTs) in the detoxification of dietary allelochemicals in the digestive gland of the predatory gastropod Cyphoma gibbosum, a generalist consumer of gorgonian corals. Controlled laboratory feeding experiments were used to investigate the influence of gorgonian diet on Cyphoma GST activity and isoform expression. Gorgonian extracts and semi-purified fractions were also screened to identify inhibitors and possible substrates of Cyphoma GSTs. In addition, we investigated the inhibitory properties of prostaglandins (PGs) structurally similar to antipredatory PGs found in high concentrations in the Caribbean gorgonian Plexaura homomalla. Cyphoma GST subunit composition was invariant and activity was constitutively high regardless of gorgonian diet. Bioassay-guided fractionation of gorgonian extracts revealed that moderately hydrophobic fractions from all eight gorgonian species examined contained putative GST substrates/inhibitors. LC-MS and NMR spectral analysis of the most inhibitory fraction from P. homomalla subsequently identified prostaglandin A2 (PGA2) as the dominant component. A similar screening of commercially available prostaglandins in series A, E, and F revealed that those prostaglandins most abundant in gorgonian tissues (e.g., PGA2) were also the most potent inhibitors. In vivo estimates of PGA2 concentration in digestive gland tissues calculated from snail grazing rates revealed that Cyphoma GSTs would be saturated with respect to PGA2 and operating at or near physiological capacity. The high, constitutive activity of Cyphoma GSTs is likely necessitated by the ubiquitous presence of GST substrates and/or inhibitors in this consumer's gorgonian diet. This generalist's GSTs may operate as ‘all-purpose’ detoxification enzymes, capable of conjugating or sequestering a broad range of lipophilic gorgonian compounds, thereby allowing this predator to exploit a range of chemically-defended prey, resulting in a competitive dietary advantage for this species.Financial support for this work was provided by the Ocean Life Institute Tropical Research Initiative Grant (WHOI) to KEW and MEH; the Robert H. Cole Endowed Ocean Ventures Fund (WHOI) to KEW; the National Undersea Research Center - Program Development Proposal (CMRC-03PRMN0103A) to KEW; Walter A. and Hope Noyes Smith, and a National Science Foundation Graduate Research Fellowship to KEW

Developing a hypothetical multi-dimensional learning progression for the nature of matter

We describe efforts toward the development of a hypothetical learning progression (HLP) for the growth of grade 7–14 students' models of the structure, behavior and properties of matter, as it relates to nanoscale science and engineering (NSE). This multi-dimensional HLP, based on empirical research and standards documents, describes how students need to incorporate and connect ideas within and across their models of atomic structure, the electrical forces that govern interactions at the nano-, molecular, and atomic scales, and information in the Periodic Table to explain a broad range of phenomena. We developed a progression from empirical data that characterizes how students currently develop their knowledge as part of the development and refinement of the HLP. We find that most students are currently at low levels in the progression, and do not perceive the connections across strands in the progression that are important for conceptual understanding. We suggest potential instructional strategies that may help students build organized and integrated knowledge structures to consolidate their understanding, ready them for new ideas in science, and help them construct understanding of emerging disciplines such as NSE, as well as traditional science disciplines. © 2009 Wiley Periodicals, Inc. J Res Sci Teach 47:687–715, 2010Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/77521/1/20324_ftp.pd

Neighbourhood ethnic density effects on behavioural and cognitive problems among young racial/ethnic minority children in the US and England: a cross-national comparison

Studies on adult racial/ethnic minority populations show that the increased concentration of racial/ethnic minorities in a neighbourhood—a so-called ethnic density effect—is associated with improved health of racial/ethnic minority residents when adjusting for area deprivation. However, this literature has focused mainly on adult populations, individual racial/ethnic groups, and single countries, with no studies focusing on children of different racial/ethnic groups or comparing across nations. This study aims to compare neighbourhood ethnic density effects on young children’s cognitive and behavioural outcomes in the US and in England. We used data from two nationally representative birth cohort studies, the US Early Childhood Longitudinal Study-Birth Cohort and the UK Millennium Cohort Study, to estimate the association between own ethnic density and behavioural and cognitive development at 5 years of age. Findings show substantial heterogeneity in ethnic density effects on child outcomes within and between the two countries, suggesting that ethnic density effects may reflect the wider social and economic context. We argue that researchers should take area deprivation into account when estimating ethnic density effects and when developing policy initiatives targeted at strengthening and improving the health and development of racial and ethnic minority children

Soft Coral Sarcophyton (Cnidaria: Anthozoa: Octocorallia) Species Diversity and Chemotypes

Research on the soft coral genus Sarcophyton extends over a wide range of fields, including marine natural products and the isolation of a number of cembranoid diterpenes. However, it is still unknown how soft corals produce this diverse array of metabolites, and the relationship between soft coral diversity and cembranoid diterpene production is not clear. In order to understand this relationship, we examined Sarcophyton specimens from Okinawa, Japan, by utilizing three methods: morphological examination of sclerites, chemotype identification, and phylogenetic examination of both Sarcophyton (utilizing mitochondrial protein-coding genes MutS homolog: msh1) and their endosymbiotic Symbiodinium spp. (utilizing nuclear internal transcribed spacer of ribosomal DNA: ITS- rDNA). Chemotypes, molecular phylogenetic clades, and sclerites of Sarcophyton trocheliophorum specimens formed a clear and distinct group, but the relationships between chemotypes, molecular phylogenetic clade types and sclerites of the most common species, Sarcophyton glaucum, was not clear. S. glaucum was divided into four clades. A characteristic chemotype was observed within one phylogenetic clade of S. glaucum. Identities of symbiotic algae Symbiodinium spp. had no apparent relation to chemotypes of Sarcophyton spp. This study demonstrates that the complex results observed for S. glaucum are due to the incomplete and complex taxonomy of this species group. Our novel method of identification should help contribute to classification and taxonomic reassessment of this diverse soft coral genus

Psychological distress during pregnancy in a multi-ethnic community: findings from the born in Bradford cohort study

Purpose: Antenatal anxiety and depression are predictive of future mental distress, which has negative effects on children. Ethnic minority women are more likely to have a lower socio-economic status (SES) but it is unclear whether SES is an independent risk factor for mental health in pregnancy. We described the association between maternal mental distress and socio-demographic factors in a multi-ethnic cohort located in an economically deprived city in the UK. Methods: We defined eight distinct ethno-language groups (total N = 8,454) and classified a threshold of distress as the 75th centile of within-group GHQ-28 scores, which we used as the outcome for univariate and multivariate logistic regression for each ethnic group and for the sample overall. Results: Financial concerns were strongly and independently associated with worse mental health for six out of the eight ethnic groups, and for the cohort overall. In some groups, factors such as working status, education and family structure were associated with worse mental health, but for others these factors were of little importance. Conclusions: The diversity between and within ethnic groups in this sample underlines the need to take into consideration individual social, migration and economic circumstances and their potential effect on mental health in ethnically diverse areas

The ε3 and ε4 Alleles of Human APOE Differentially Affect Tau Phosphorylation in Hyperinsulinemic and Pioglitazone Treated Mice

Impaired insulin signalling is increasingly thought to contribute to Alzheimer's disease (AD). The ε4 isoform of the APOE gene is the greatest genetic risk factor for sporadic, late onset AD, and is also associated with risk for type 2 diabetes mellitus (T2DM). Neuropathological studies reported the highest number of AD lesions in brain tissue of ε4 diabetic patients. However other studies assessing AD pathology amongst the diabetic population have produced conflicting reports and have failed to show an increase in AD-related pathology in diabetic brain. The thiazolidinediones (TZDs), peroxisome proliferator-activated receptor gamma agonists, are peripheral insulin sensitisers used to treat T2DM. The TZD, pioglitazone, improved memory and cognitive functions in mild to moderate AD patients. Since it is not yet clear how apoE isoforms influence the development of T2DM and its progression to AD, we investigated amyloid beta and tau pathology in APOE knockout mice, carrying human APOEε3 or ε4 transgenes after diet-induced insulin resistance with and without pioglitazone treatment.Male APOE knockout, APOEε3-transgenic and APOEε4-transgenic mice, together with background strain C57BL6 mice were kept on a high fat diet (HFD) or low fat diet (LFD) for 32 weeks, or were all fed HFD for 32 weeks and during the final 3 weeks animals were treated with pioglitazone or vehicle.All HFD animals developed hyperglycaemia with elevated plasma insulin. Tau phosphorylation was reduced at 3 epitopes (Ser396, Ser202/Thr205 and Thr231) in all HFD, compared to LFD, animals independent of APOE genotype. The introduction of pioglitazone to HFD animals led to a significant reduction in tau phosphorylation at the Ser202/Thr205 epitope in APOEε3 animals only. We found no changes in APP processing however the levels of soluble amyloid beta 40 was reduced in APOE knockout animals treated with pioglitazone