Benzobistriazinones and related heterocyclic ring systems as potent, orally bioavailable positive allosteric AMPA receptor modulators

AMPA receptors (AMPARs) are an important therapeutic target in the CNS. A series of substituted benzobistriazinone, benzobispyrimidinone and related derivatives have been prepared with high potency and selectivity for the allosteric binding site of AMPARs. Further improvements have been made to previously reported series of positive AMPAR modulators and these compounds exhibit excellent in vivo activity and improved in vivo metabolic stability with up to 100% oral bioavailability in rat.     

Effects of cord serum insulin, IGF-II, IGFBP-2, IL-6 and cortisol concentrations on human birth weight and length: pilot study

Background

The IGF system is recognised to be important for fetal growth. We previously described increased Insulin-like growth factor binding protein (IGFBP)-2 cord serum concentrations in intra-uterine growth retardation (IUGR) compared with appropriate for gestational age (AGA) newborns, and a positive relationship of IGFBP-2 with Interleukin (IL)-6. The role of cortisol in the fetus at birth is largely unknown, and interactions among peptides are their real effect on birth size is unknown. Furthermore, almost all studies have previously assayed peptides in serum several years after birth, and follow-up data from pregnancy are always lacking. This study aimed at establishing and clarifying the effect of cord serum insulin, IGF-II, IGFBP-2, cortisol and IL-6 concentrations on birth length and weight.

Methods

23 IUGR and 37 AGA subjects were followed up from the beginning of pregnancy, and were of comparable gestational age. Insulin, IGF-II, IGFBP-2, cortisol and IL-6 concentrations were assayed in cord serum at birth, and a multiple regression model was designed and applied to assess which were the significant biochemical determinants of birth size.

Results

Insulin, cortisol, and IL-6, showed similar concentrations in IUGR and AGA as previously described, whereas IGF-II was lower, and IGFBP-2 increased in IUGR compared with AGA. IGF-II serum concentration was found to have a significant positive effect on both birth length (r:^:0.546; p: 0.001) and weight (r:0.679; p: 0.0001). IGFBP-2 had a near significant negative effect on both birth weight (r:−0.342; p: 0.05) and length (r:−0.372; p:0.03).

Conclusion

IGF-II cord serum concentration was shown to have a significant positive effect on both birth length and weight, whereas IGFBP-2 had a significant negative effect. Insulin, cortisol, and IL-6 cord serum concentrations had no significant effect on birth size.     

A mitochondrial species identification assay for Australian blacktip sharks (Carcharhinus tilstoni, C. limbatus and C. amblyrhynchoides) using real-time PCR and high-resolution melt analysis

Tropical Australian shark fisheries target two morphologically indistinguishable blacktip sharks, the Australian blacktip (Carcharhinus tilstoni) and the common blacktip (C. limbatus). Their relative contributions to northern and eastern Australian coastal fisheries are unclear because of species identification difficulties. The two species differ in their number of precaudal vertebrae, which is difficult and time consuming to obtain in the field. But, the two species can be distinguished genetically with diagnostic mutations in their mitochondrial DNA ND4 gene. A third closely related sister species, the graceful shark C. amblyrhynchoides, can also be distinguished by species‐specific mutations in this gene. DNA sequencing is an effective diagnostic tool, but is relatively expensive and time consuming. In contrast, real‐time high‐resolution melt (HRM) PCR assays are rapid and relatively inexpensive. These assays amplify regions of DNA with species‐specific genetic mutations that result in PCR products with unique melt profiles. A real‐time HRM PCR species‐diagnostic assay (RT‐HRM‐PCR) has been developed based on the mtDNA ND4 gene for rapid typing of C. tilstoni, C. limbatus and C. amblyrhynchoides. The assay was developed using ND4 sequences from 66 C. tilstoni, 33. C. limbatus and five C. amblyrhynchoides collected from Indonesia and Australian states and territories; Western Australia, the Northern Territory, Queensland and New South Wales. The assay was shown to be 100% accurate on 160 unknown blacktip shark tissue samples by full mtDNA ND4 sequencing.     

A collection of INDEL markers for map-based cloning in seven Arabidopsis accessions

The availability of a comprehensive set of resources including an entire annotated reference genome, sequenced alternative accessions, and a multitude of marker systems makes Arabidopsis thaliana an ideal platform for genetic mapping. PCR markers based on INsertions/DELetions (INDELs) are currently the most frequently used polymorphisms. For the most commonly used mapping combination, Columbia×Landsberg erecta (Col-0×Ler-0), the Cereon polymorphism database is a valuable resource for the generation of polymorphic markers. However, because the number of markers available in public databases for accessions other than Col-0 and Ler-0 is extremely low, mapping using other accessions is far from straightforward. This issue arose while cloning mutations in the Wassilewskija (Ws-4) background. In this work, approaches are described for marker generation in Ws-4 x Col-0. Complementary strategies were employed to generate 229 INDEL markers. Firstly, existing Col-0/Ler-0 Cereon predicted polymorphisms were mined for transferability to Ws-4. Secondly, Ws-0 ecotype Illumina sequence data were analyzed to identify INDELs that could be used for the development of PCR-based markers for Col-0 and Ws-4. Finally, shotgun sequencing allowed the identification of INDELs directly between Col-0 and Ws-4. The polymorphism of the 229 markers was assessed in seven widely used Arabidopsis accessions, and PCR markers that allow a clear distinction between the diverged Ws-0 and Ws-4 accessions are detailed. The utility of the markers was demonstrated by mapping more than 35 mutations in a Col-0×Ws-4 combination, an example of which is presented here. The potential contribution of next generation sequencing technologies to more traditional map-based cloning is discussed.     

Incident radiation and the allocation of nitrogen within Arctic plant canopies: implications for predicting gross primary productivity

Arctic vegetation is characterized by high spatial variability in plant functional type (PFT) composition and gross primary productivity (P). Despite this variability, the two main drivers of P in sub‐Arctic tundra are leaf area index (L_T) and total foliar nitrogen (N_T). L_T and N_T have been shown to be tightly coupled across PFTs in sub‐Arctic tundra vegetation, which simplifies up‐scaling by allowing quantification of the main drivers of P from remotely sensed L_T. Our objective was to test the L_T–N_T relationship across multiple Arctic latitudes and to assess L_T as a predictor of P for the pan‐Arctic. Including PFT‐specific parameters in models of L_T–N_T coupling provided only incremental improvements in model fit, but significant improvements were gained from including site‐specific parameters. The degree of curvature in the L_T–N_T relationship, controlled by a fitted canopy nitrogen extinction co‐efficient, was negatively related to average levels of diffuse radiation at a site. This is consistent with theoretical predictions of more uniform vertical canopy N distributions under diffuse light conditions. Higher latitude sites had higher average leaf N content by mass (N_M), and we show for the first time that L_T–N_T coupling is achieved across latitudes via canopy‐scale trade‐offs between N_M and leaf mass per unit leaf area (L_M). Site‐specific parameters provided small but significant improvements in models of P based on L_T and moss cover. Our results suggest that differences in L_T–N_T coupling between sites could be used to improve pan‐Arctic models of P and we provide unique evidence that prevailing radiation conditions can significantly affect N allocation over regional scales.     

IL4 and IL5: The role of two multifunctional cytokines and their place in the network of cytokine interactions

The DNAX Research Institute of Molecular and Cellular Biology is supported by Schering-Plough Corporation.     

Five QTL hotspots for yield in short rotation coppice bioenergy poplar: The Poplar Biomass Loci

Background  

Concern over land use for non-food bioenergy crops requires breeding programmes that focus on producing biomass on the minimum amount of land that is economically-viable. To achieve this, the maximum potential yield per hectare is a key target for improvement. For long lived tree species, such as poplar, this requires an understanding of the traits that contribute to biomass production and their genetic control. An important aspect of this for long lived plants is an understanding of genetic interactions at different developmental stages, i.e. how genes or genetic regions impact on yield over time.     

Genomic Analyses of the Microsporidian Nosema ceranae,an Emergent Pathogen of Honey Bees

Recent steep declines in honey bee health have severely impacted the beekeeping industry, presenting new risks for agricultural commodities that depend on insect pollination. Honey bee declines could reflect increased pressures from parasites and pathogens. The incidence of the microsporidian pathogen Nosema ceranae has increased significantly in the past decade. Here we present a draft assembly (7.86 MB) of the N. ceranae genome derived from pyrosequence data, including initial gene models and genomic comparisons with other members of this highly derived fungal lineage. N. ceranae has a strongly AT-biased genome (74% A+T) and a diversity of repetitive elements, complicating the assembly. Of 2,614 predicted protein-coding sequences, we conservatively estimate that 1,366 have homologs in the microsporidian Encephalitozoon cuniculi, the most closely related published genome sequence. We identify genes conserved among microsporidia that lack clear homology outside this group, which are of special interest as potential virulence factors in this group of obligate parasites. A substantial fraction of the diminutive N. ceranae proteome consists of novel and transposable-element proteins. For a majority of well-supported gene models, a conserved sense-strand motif can be found within 15 bases upstream of the start codon; a previously uncharacterized version of this motif is also present in E. cuniculi. These comparisons provide insight into the architecture, regulation, and evolution of microsporidian genomes, and will drive investigations into honey bee–Nosema interactions.     

Measuring healthy behaviours using the stages of change model: an investigation into the physical activity and nutrition behaviours of Australian miners

Background

Obesity is one of the fastest growing modern day epidemics affecting preventable disease and premature deaths. Healthy lifestyle behaviours, such as physical activity and nutritional consumption, have been shown to reduce the likelihood of obesity and obesity related health risks. Originally designed for measurement of unhealthy behaviours, the Stages of Change model, describes ‘precontemplators’ as individuals who engage in the unhealthy behaviour, are unaware that their behaviour is problematic, and are resistant to change. The aim of this study was to refine and assess the measures of the Stages of Change model in order to achieve a concise and reliable classification of precontemplators, in the context of healthy behaviours.

Methods

Eight hundred and ninety-seven employees participated in a health survey measuring current health behaviours and stage of change. This study compared a traditional precontemplation measure to a modified version in the assessment of two healthy behaviours: physical activity and fruit and vegetable consumption.

Results

The modified measure was more accurate and captured fewer individuals currently meeting the guideline for both physical activity and nutrition, compared to the traditional measure of stages of change. However, across all stages of change, the measure incorrectly classified some employees with regards to meeting health guidelines.

Conclusions

When applied to healthy behaviours, the stages of change measure for precontemplation should be further refined to reflect knowledge that the behaviour is unhealthy, and apathy to change. Additionally, measures should define health guidelines to increase reliable classification across all stages of change. The findings can be applied to inform the design and implementation of health promotion strategies targeting obesity related lifestyle behaviours in the general population.