EFFECT OF LIGHT CYCLE ON DIATOM FATTY ACID COMPOSITION AND QUANTITATIVE MORPHOLOGY1

Quantitative cytological and fatty acid composition was determined for the diatom Cyclotella meneghiniana Kütz, Data from four separate experiments were examined to elucidate changes that may occur with respect to daily photoperiod. Overall, fatty acid composition is similar to that reported for other diatoms with the exception that the C16 fatty acids constitute approximately 70% of all fatty acids. The major fatty acids are C14:0, 16:1, 16:0, 18:0, and 20:5. Fatty acids that are present in minor amounts are iso-14:0, iso-15:0, 15:0, 17:0, 18:4, 18:2, 18:1, 19:0, 20:0, 22:0, and 23:0. Cytological composition is similar to that previously reported with the chloroplast and vacuole being the largest compartments within the cell. Changes in both cytological and fatty acid composition were studied with respect to the light / dark cycle. Chloroplast and lipid relative volume are greatest during the early part of the dark period. Nuclear relative volume is lowest in the dark and increases throughout the light period. Total unsaturated fatty acids, including the C20:5 fatty acid, are lowest in the early part of the light period and highest in the dark. The sum of the C16 fatty acids remains constant at 70% of total fatty acids in the cells throughout the light/dark cycle, although percent composition of these two fatty acids shifts. The data suggest that cyclical changes occur in both quantitative morphology and fatty acids composition with respect to daily photoperiod. The cells, although not rigidly synchronized, most likely divide in the latter part of the dark period or in the first hours of the light period. Lipids increase dramatically in the dark. The ecological implications of lipid storage are discussed in relation to lipophilic toxicants.     

Update of MmtDB: a Metazoa mitochondrial DNA variants database.

The present paper describes the improvements in MmtDB, a specialised database designed to collect Metazoa mitochondrial DNA variants. Priority in the data collection has been given to Metazoa for which a large amount of variants is available, e.g., for humans. Starting from the sequences available in the Nucleotide Sequence Databases, the redundant sequences have been removed and new sequences from other sources have been added. Value-added information is associated to each variant sequence, e.g., analysed region, experimental method, tissue and cell lines, population data, sex, age, family code and information about the variation events (nucleotide position, involved gene, restriction site gain or loss). Cross-references are introduced to the EMBL Data Library, as well as an internal cross-referencing among MmtDB entries according to tissual, heteroplasmic, familiar and aplotypical correlation. Furthermore MmtDB has a new section, AMmtDB: Aligned Metazoan mitochondrial biosequences. MmtDB can be accessed through the World Wide Web at URL http://WWW.ba.cnr.it/[symbol: see text]areamt08/MmtDBWWW.htm     

Hybridization Capture Using RAD Probes (hyRAD), a New Tool for Performing Genomic Analyses on Collection Specimens

In the recent years, many protocols aimed at reproducibly sequencing reduced-genome subsets in non-model organisms have been published. Among them, RAD-sequencing is one of the most widely used. It relies on digesting DNA with specific restriction enzymes and performing size selection on the resulting fragments. Despite its acknowledged utility, this method is of limited use with degraded DNA samples, such as those isolated from museum specimens, as these samples are less likely to harbor fragments long enough to comprise two restriction sites making possible ligation of the adapter sequences (in the case of double-digest RAD) or performing size selection of the resulting fragments (in the case of single-digest RAD). Here, we address these limitations by presenting a novel method called hybridization RAD (hyRAD). In this approach, biotinylated RAD fragments, covering a random fraction of the genome, are used as baits for capturing homologous fragments from genomic shotgun sequencing libraries. This simple and cost-effective approach allows sequencing of orthologous loci even from highly degraded DNA samples, opening new avenues of research in the field of museum genomics. Not relying on the restriction site presence, it improves among-sample loci coverage. In a trial study, hyRAD allowed us to obtain a large set of orthologous loci from fresh and museum samples from a non-model butterfly species, with a high proportion of single nucleotide polymorphisms present in all eight analyzed specimens, including 58-year-old museum samples. The utility of the method was further validated using 49 museum and fresh samples of a Palearctic grasshopper species for which the spatial genetic structure was previously assessed using mtDNA amplicons. The application of the method is eventually discussed in a wider context. As it does not rely on the restriction site presence, it is therefore not sensitive to among-sample loci polymorphisms in the restriction sites that usually causes loci dropout. This should enable the application of hyRAD to analyses at broader evolutionary scales.     

Time trends in symptoms of mental illness in children and adolescents in Canada

Background:

Existing research and media reports convey conflicting impressions of trends in the prevalence of mental illness. We sought to investigate trends in the prevalence of symptoms of mental illness in a large population-based cohort of Canadian children and adolescents.

Methods:

We obtained population-based data from the National Longitudinal Survey of Children and Youth. Every 2 years, participants completed self-reported measures of mental illness indicators, including conduct disorder, hyperactivity, indirect aggression, suicidal behaviour, and depression and anxiety. We analyzed trends in mean scores over time using linear regression.

Results:

We evaluated 11 725 participants aged 10–11 years from cycles 1 (1994/95) through 6 (2004/05), 10 574 aged 12–13 years from cycles 2 (1996/97) through 7 (2006/07), and 9835 aged 14–15 years from cycles 3 (1998/99) through 8 (2008/09). The distribution of scores on depression and anxiety, conduct and indirect aggression scales remained stable or showed small decreases over time for participants of all ages. The mean hyperactivity score increased over time in participants aged 10–11 years (change per 2-year cycle: 0.16, 95% CI 0.02 to 0.12) and those aged 12–13 years (0.13, 95% CI 0.09 to 0.18). Over time, fewer participants aged 12–13 years (0.40% per cycle, 95% CI −0.78 to −0.07) and aged 14–15 years (0.56% per cycle, 95% CI −0.91 to −0.23) reported attempting suicide in the previous 12 months.

Interpretation:

With the exception of hyperactivity, the prevalence of symptoms of mental illness in Canadian children and adolescents has remained relatively stable from 1994/95 to 2008/09. Conflicting reports of escalating rates of mental illness in Canada may be explained by differing methodologies between studies, an increase in treatment-seeking behaviour, or changes in diagnostic criteria or practices.Popular media tends to perpetuate the idea that the prevalence of mental disorders is increasing. However, research supporting this position has been inconsistent. Several studies have shown increases in recent years,^1,2 including among adolescents.^3–7 These studies have included parent reports from population samples^3,4 and physician diagnoses from electronic medical records.⁶ In contrast, other studies have reported that the rates of mental illness have decreased or remained stable over time.^8–11 To appropriately plan services and policies, a proper understanding of trends in adolescent mental health is crucial. This is particularly relevant in Canada, where policy regarding mental health is currently being reconsidered under such frameworks as the Mental Health Commission of Canada’s 2012 mental health strategy, “Changing Directions, Changing Lives.”¹²Many methodologic challenges make it difficult to draw conclusions regarding trends in mental illness. The conflicting reports of changing rates of mental illness in the population may be partially explained by changes in diagnostic criteria, differences in assessment methods or variations in official reporting practices.⁴ For example, one study found that while diagnosed depression decreased, the incidence of depressive symptoms noted by health professionals increased by threefold from 1996 to 2006.¹³ These changes may not represent a real change in the rates of depression or depressive symptoms, but rather a change in clinical practice.Canada’s National Longitudinal Survey of Children and Youth repeatedly included a series of questions from 1994/95 to 2008/09 relevant to many aspects of mental health in children and adolescents, including assessments of conduct disorder, hyperactivity, aggression, depression and anxiety, and suicidal thinking and behaviour. Our primary objective was to investigate trends in the prevalence of symptoms in these areas in 3 specific age groups: 10–11, 12–13 and 14–15 years.     

alpha-Synuclein protects SH-SY5Y cells from dopamine toxicity

Dopaminergic human neuroblastoma SH-SY5Y cells were stably transformed to increase expression of alpha-synuclein, a Parkinson's disease-related protein. Transformed cells were more resistant to oxidative insults, showing a cytoprotective role of alpha-synuclein. The expression of redox chaperonins (DJ-1, HSP70, and 14-3-3) was evaluated by Western blotting. Expression of alpha-synuclein reduced HSP70 levels even in the presence of dopamine, with a twofold increase of DJ-1 in the absence of oxidants. DJ-1 is significantly reduced by dopamine, and even more by dopamine and Cu(II). Increased alpha-synuclein expression did not affect 14-3-3, although dopamine increased its level by 60% in wild-type cells. alpha-Synuclein not only upregulated DJ-1, but also shifted all DJ-1 forms to a single spot at pI=5.7 not observed in wild-type cells. Dopamine gradually restored the distribution of DJ-1 forms to a situation similar to wild-type cells, with the form at pI=6.1 progressively enriched under oxidative conditions.     

Characterization of three members of the Arabidopsis carotenoid cleavage dioxygenase family demonstrates the divergent roles of this multifunctional enzyme family

Arabidopsis thaliana has nine genes that constitute a family of putative carotenoid cleavage dioxygenases (CCDs). While five members of the family are believed to be involved in synthesis of the phytohormone abscisic acid, the functions of the other four enzymes are less clear. Recently two of the enzymes, CCD7/MAX3 and CCD8/MAX4, have been implicated in synthesis of a novel apocarotenoid hormone that controls lateral shoot growth. Here, we report on the molecular and genetic interactions between CCD1, CCD7/MAX3 and CCD8/MAX4. CCD1 distinguishes itself from other reported CCDs as being the only member not targeted to the plastid. Unlike ccd7/max3 and ccd8/max4, both characterized as having highly branched phenotypes, ccd1 loss-of-function mutants are indistinguishable from wild-type plants. Thus, even though CCD1 has similar enzymatic activity to CCD7/MAX3, it does not have a role in synthesis of the lateral shoot growth inhibitor. Rather, it may have a role in synthesis of apocarotenoid flavor and aroma volatiles, especially in maturing seeds where loss of function leads to significantly higher carotenoid levels.     

A Simple Method for Assessing Free Brain/Free Plasma Ratios Using an In Vitro Model of the Blood Brain Barrier

Historically, the focus has been to use in vitro BBB models to optimize rate of drug delivery to the CNS, whereas total in vivo brain/plasma ratios have been used for optimizing extent. However, these two parameters do not necessarily show good correlations with receptor occupancy data or other pharmacological readouts. In line with the free drug hypothesis, the use of unbound brain concentrations (Cu,br) has been shown to provide the best correlations with pharmacological data. However, typically the determination of this parameter requires microdialysis, a technique not ideally suited for screening in early drug development. Alternative, and less resource-demanding methodologies to determine Cu,br employ either equilibrium dialysis of brain homogenates or incubations of brain slices in buffer to determine fraction unbound brain (fu,br), which is subsequently multiplied by the total brain concentration to yield Cu,br. To determine Cu,br/Cu,pl ratios this way, still requires both in vitro and in vivo experiments that are quite time consuming. The main objective of this study was to explore the possibility to directly generate Cu,br/Cu,pl ratios in a single in vitro model of the BBB, using a co-culture of brain capillary endothelial and glial cells in an attempt to mimick the in vivo situation, thereby greatly simplifying existing experimental procedures. Comparison to microdialysis brain concentration profiles demonstrates the possibility to estimate brain exposure over time in the BBB model. A stronger correlation was found between in vitro Cu,br/Cu,pl ratios and in vivo Cu,br/Cu,pl obtained using fu,br from brain slice than with fu,br from brain homogenate for a set of 30 drugs. Overall, Cu,br/Cu,pl ratios were successfully predicted in vitro for 88% of the 92 studied compounds. This result supports the possibility to use this methodology for identifying compounds with a desirable in vivo response in the CNS early on in the drug discovery process.     

Characters of compost teas from different sources and their suppressive effect on fungal phytopathogens

Compost teas (CT) are fermented watery extracts of composted materials that are used to control plant diseases and on crop fertilization. In this work, aerated (ACT) and non-aerated compost teas (NCT) were obtained from four different composts: spent mushroom substrate compost, grape marc compost, greenhouse horticultural crop residues compost, and vermicompost. Physico-chemical and microbiological analysis were carried out to determine their properties. In vitro assays were performed to assess their suppressive effect on the mycelial growth of eight fungal phytopathogens. In vivo trials aimed to assess their effect on gummy stem blight (Didymella bryonae) and powdery mildew (Podosphaera fusca) in melon plants. Results showed that ACT and NCT filtrates inhibited the in vitro growth of all tested pathogens while autoclaved CT did not completely lose their inhibitory effect, and CT sterilized by microfiltration had no effect on the pathogen growth. The severity of powdery mildew was highly reduced by ACT and NCT from all sources, though in gummy stem blight assay only a delay in disease development was observed. In general, all compost teas showed a high level of microbial populations and nutrients. Results suggest that the efficacy of ACT and NCT firstly depend on the microbiota present in them. We consider compost teas from the four tested sources as a viable way to manage plant diseases and crop fertilization, throughout its integration in pest management programs and fertirrigation systems under different dilution rates.