Bioactivity of some natural products against the cowpea storage weevil Callosobruchus Maculatus L.

This work was carried out to evaluate the insecticidal activity of extracts from 14 plant species of Ghanaian origin against Callosobruchus maculatus adults under laboratory conditions. Phytochemical screenings of the 14 plant species were performed using standard laboratory methods to determine the types of secondary natural products present in the plant species. Aqueous, petroleum ether and ethanolic extracts of the 14 plant species were screened for their bioactivity against Artemia selina in a brine shrimp lethality test. The pesticidal activities of the plant products against Callosobruchus maculatus were investigated in laboratory experiments. Contact toxicity (filter paper impregnation method), grain treatment with plant products for insect population and mortality assays, and also effect of the plant products on grain damage were obtained and compared with Actellic Super EC (synthetic insecticide) during a three-month observation period. Cymbopogan winterianus oil caused the highest mortality, least damage to grains and the highest antireproductive effect. All grain-protective properties of the natural products were concentration and time dependent. Extracts of Cassia alata showed good potential in protecting cowpea and gave better protection than the powder, whilst Griffornia simplicifolia did not significantly protect the grains. Phytochemical investigations of the plants revealed the presence of terpenes, saponins, tannins, glycosides, alkaloids, anthraquinones and flavonoids. The presence of some of these compounds may be responsible for the pesticidal action of the plants.Keywords: Phytochemical screening, Extracts, Insecticidal, Brine shrimp lethality test, Cowpea

Identification of semiochemicals from cowpea, Vigna unguiculata, for low-input management of the Legume Pod Borer, Maruca vitrata

Cowpea, Vigna unguiculata L. Walp. (Fabaceae), is one of the most important food legumes grown on the African continent, as it provides an affordable source of dietary protein. Yields of cowpea are significantly reduced through damage by legume pod-borer, Maruca vitrata (Lepidoptera: Crambidae), caterpillars to flowers, tender leaves and pods. Semiochemical-based strategies are considered as environmentally benign and affordable for pest management, particularly on smallholder farms. In this study, we investigated the importance of cowpea flower volatiles as host location cues for egg-laying M. vitrata, and herbivore-induced plant volatiles (HIPVs) as M. vitrata repellents and natural enemy (Apanteles taragamae and Phanerotoma syleptae parasitoid) attractants. In oviposition choice assays, M. vitrata laid more eggs on flowering cowpea plants than non-flowering plants. Coupled gas chromatography-electrophysiology (GC-EAG) analysis using the antennae of female M. vitrata and an extract of flower volatiles collected by dynamic headspace collection revealed the presence of five EAG-active components that were identified by coupled GC-mass spectrometry (GC-MS) analysis as benzaldehyde, benzyl alcohol, acetophenone, a vinylbenzaldehyde isomer and (E)-cinnamaldehyde. A synthetic blend of the identified compounds, prepared using 3-vinylbenzaldehyde, induced M. vitrata to lay as many eggs on non-flowering cowpea as on flowering plants. The moths also preferred laying eggs on intact plants compared to M. vitrata-infested plants. As the emission of EAG-active floral compounds was determined to be lower in the headspace of infested cowpea flowers, the role of HIPVs emitted by M. vitrata-damaged leaves was also investigated. Of the compounds induced by larval damage, (E)-DMNT, indole, n-hexyl acetate, 1-octen-3-ol and linalool were shown by GC-EAG to possess electrophysiological activity. A synthetic blend of the EAG-active compounds, using racemic 1-octen-3-ol and linalool, significantly reduced egg numbers on flowering cowpea. Larval and egg parasitoids, i.e. A. taragamae and Ph. syleptae, respectively, of M. vitrata both preferred the Y-tube olfactometer arm treated with synthetic (E)-DMNT, whereas preference for racemic linalool and (E)-nerolidol was dose-dependent in A. taragamae. Our results provide the platform for the development of future semiochemical-based pest management strategies against M. vitrata on smallholder farms in West Africa

(2R,5S)-theaspirane identified as the kairomone for the banana weevil, Cosmopolites sordidus, from attractive senesced leaves of the host banana, Musa spp.

The principal active component produced by highly attractive senesced host banana leaves, Musa spp., for the banana weevil, Cosmopolites sordidus, is shown by coupled gas chromatography‐electroantennography (GC‐EAG), coupled GC‐mass spectrometry (GC‐MS), chemical synthesis and coupled enantioselective (chiral) GC‐EAG to be (2R,5S)‐theaspirane. In laboratory behaviour tests, the synthetic compound is as attractive as natural host leaf material and presents a new opportunity for pest control

Cardiac myosin binding protein-C variants in paediatric-onset hypertrophic cardiomyopathy: natural history and clinical outcomes

Background: Variants in the cardiac myosin-binding protein C gene (MYBPC3) are a common cause of hypertrophic cardiomyopathy (HCM) in adults and have been associated with late-onset disease, but there are limited data on their role in paediatric-onset HCM. The objective of this study was to describe natural history and clinical outcomes in a large cohort of children with HCM and pathogenic/likely pathogenic (P/LP) MYBPC3 variants. / Methods and results: Longitudinal data from 62 consecutive patients diagnosed with HCM under 18 years of age and carrying at least one P/LP MYBPC3 variant were collected from a single specialist referral centre. The primary patient outcome was a major adverse cardiac event (MACE). Median age at diagnosis was 10 (IQR: 2–14) years, with 12 patients (19.4%) diagnosed in infancy. Forty-seven (75%) were boy and 31 (50%) were probands. Median length of follow-up was 3.1 (IQR: 1.6–6.9) years. Nine patients (14.5%) experienced an MACE during follow-up and five (8%) died. Twenty patients (32.3%) had evidence of ventricular arrhythmia, including 6 patients (9.7%) presenting with out-of-hospital cardiac arrest. Five-year freedom from MACE for those with a single or two MYBPC3 variants was 95.2% (95% CI: 78.6% to 98.5%) and 68.4% (95% CI: 40.6% to 88.9%), respectively (HR 4.65, 95% CI: 1.16 to 18.66, p=0.03). / Conclusions: MYBPC3 variants can cause childhood-onset disease, which is frequently associated with life-threatening ventricular arrhythmia. Clinical outcomes in this cohort vary substantially from aetiologically and genetically mixed paediatric HCM cohorts described previously, highlighting the importance of identifying specific genetic subtypes for clinical management of childhood HCM

Macromolecular Traits in the African Rice Oryza glaberrima and in Glaberrima/Sativa Crosses, and Their Relevance to Processing

Molecular properties of proteins and starch were investigated in 2 accessions of Oryza glaberrima and Oryza sativa, and in one NERICA cross between the 2 species, to assess traits that could be relevant to transformation into specific foods. Protein nature and organization in O. glaberrima were different from those in O. sativa and in NERICA. Despite the similar cysteine content in all samples, thiol accessibility in O. glaberrima proteins was higher than in NERICA or in O. sativa. Inter-protein disulphide bonds were important for the formation of protein aggregates in O. glaberrima, whereas non-covalent protein-protein interactions were relevant in NERICA and O. sativa. DSC and NMR studies indicated only minor differences in the structure of starch in these species, as also made evident by their microstructural features. Nevertheless, starch gelatinization in O. glaberrima was very different from what was observed in O. sativa and NERICA. The content of soluble species in gelatinized starch from the various species in the presence/absence of treatments with specific enzymes indicated that release of small starch breakdown products was lowest in O. glaberrima, in particular from the amylopectin component. These findings may explain the low glycemic index of O. glaberrima, and provide a rationale for extending the use of O. glaberrima in the production of specific rice-based products, thus improving the economic value and the market appeal of African crops

Map-A-Mole: greenspace area influences the presence and abundance of the European mole Talpa europaea in urban habitats

The European mole Talpa europaea is common across much of Britain. It has a unique fossorial lifestyle, and evidence of its presence is readily identified through the presence of characteristic molehills. Although molehills are often a common sight in urban greenspaces, moles are remarkably understudied, with very few studies to date exploring the urban ecology of moles. Here, we investigate if factors such as greenspace (largely urban parks and playing fields) area, intensity of management, distance to nearest patch, amount of time the patch had been isolated from other green patches, and the amount of urbanization (constructed surfaces) surrounding the patch, influence the distribution and abundance of urban moles. Mole signs (hills and surface runs) were counted in all discrete urban greenspaces (excluding domestic gardens and one private golf course) within an 89.5 km2 area in the UK town of Reading. We found that 17 out of 59 surveyed sites contained moles, with their presence being recorded in greenspaces with a minimum patch area of approximately 0.1 km2 (10 ha). Where present, the abundance of mole territories in the greenspaces was associated with both the area of greenspace and degree of urbanization within 150 m of the patch boundary. While the former was not surprising, the latter outcome may be a consequence of sites with an increased risk of flooding being home to fewer moles, and the surrounding area is also less likely to be built upon. This case study highlights how choices made in designing urban green infrastructure will determine which species survive in urban areas long into the future

Analysis of anti-RNA polymerase III antibody positive systemic sclerosis suggests altered GPATCH2L and CTNND2 expression in scleroderma renal crisis

OBJECTIVE: Scleroderma renal crisis (SRC) is a life-threatening complication of systemic sclerosis (SSc) strongly associated with anti RNA polymerase III antibody (ARA) autoantibodies. We explore genetic susceptibility and altered protein expression in renal biopsy specimens in ARA positive SRC. METHODS: ARA-positive patients (n=99) with at least 5 years' follow-up (49% with a history of SRC) were selected from a well-characterised SSc cohort (n=2254). Cases were genotyped using the Illumina Human Omni-express chip. Based on initial regression analysis, nine SNPs were chosen for validation in a separate cohort of 256 ARA+ patients (40 with SRC). Immunostaining of tissue sections from SRC or control kidney was used to quantify expression of candidate proteins based upon genetic analysis of the discovery cohort. RESULTS: Analysis of 641,489 SNPs suggested association of POU2F1 (rs2093658; 1.98x10-5), CTNND2 (rs1859082; p=7.14 x 10-5), HECW2 (rs16849716; p=1.2 x 10-4) and GPATCH2L (rs935332; p=4.92 x 10-5) with SRC. Furthermore, the validation cohort showed an association between rs935332 within the GPATCH2L region, with SRC (p=0.025). Immunostaining of renal biopsy sections showed increased tubular expression of GPATCH2L (p=0.026), and glomerular expression of CTNND2 (p=0.026) in SRC samples (n=8) compared with normal human kidney controls (n=8), despite absence of any genetic replication for the associated SNP. CONCLUSION: Increased expression of two candidate proteins GPATCH2L and CTNND2 in SRC compared with control kidney suggests a potential role in pathogenesis of SRC. For GPATCH2L this may reflect genetic susceptibility in ARA positive SSc based upon 2 independent cohorts

Expanding the horizons of microRNA bioinformatics

MicroRNA regulation of key biological and developmental pathways is a rapidly expanding area of research, accompanied by vast amounts of experimental data. This data, however, is not widely available in bioinformatic resources, making it difficult for researchers to find and analyse microRNA-related experimental data and define further research projects. We are addressing this problem by providing two new bioinformatics datasets that contain experimentally verified functional information for mammalian microRNAs involved in cardiovascular-relevant, and other, processes. To date, our resource provides over 3,900 Gene Ontology annotations associated with almost 500 miRNAs from human, mouse and rat and over 2,200 experimentally validated miRNA:target interactions. We illustrate how this resource can be used to create miRNA-focused interaction networks with a biological context using the known biological role of miRNAs and the mRNAs they regulate, enabling discovery of associations between gene products, biological pathways and, ultimately, diseases. This data will be crucial in advancing the field of microRNA bioinformatics and will establish consistent datasets for reproducible functional analysis of microRNAs across all biological research areas

Network Inference Algorithms Elucidate Nrf2 Regulation of Mouse Lung Oxidative Stress

A variety of cardiovascular, neurological, and neoplastic conditions have been associated with oxidative stress, i.e., conditions under which levels of reactive oxygen species (ROS) are elevated over significant periods. Nuclear factor erythroid 2-related factor (Nrf2) regulates the transcription of several gene products involved in the protective response to oxidative stress. The transcriptional regulatory and signaling relationships linking gene products involved in the response to oxidative stress are, currently, only partially resolved. Microarray data constitute RNA abundance measures representing gene expression patterns. In some cases, these patterns can identify the molecular interactions of gene products. They can be, in effect, proxies for protein–protein and protein–DNA interactions. Traditional techniques used for clustering coregulated genes on high-throughput gene arrays are rarely capable of distinguishing between direct transcriptional regulatory interactions and indirect ones. In this study, newly developed information-theoretic algorithms that employ the concept of mutual information were used: the Algorithm for the Reconstruction of Accurate Cellular Networks (ARACNE), and Context Likelihood of Relatedness (CLR). These algorithms captured dependencies in the gene expression profiles of the mouse lung, allowing the regulatory effect of Nrf2 in response to oxidative stress to be determined more precisely. In addition, a characterization of promoter sequences of Nrf2 regulatory targets was conducted using a Support Vector Machine classification algorithm to corroborate ARACNE and CLR predictions. Inferred networks were analyzed, compared, and integrated using the Collective Analysis of Biological Interaction Networks (CABIN) plug-in of Cytoscape. Using the two network inference algorithms and one machine learning algorithm, a number of both previously known and novel targets of Nrf2 transcriptional activation were identified. Genes predicted as novel Nrf2 targets include Atf1, Srxn1, Prnp, Sod2, Als2, Nfkbib, and Ppp1r15b. Furthermore, microarray and quantitative RT-PCR experiments following cigarette-smoke-induced oxidative stress in Nrf2+/+ and Nrf2−/− mouse lung affirmed many of the predictions made. Several new potential feed-forward regulatory loops involving Nrf2, Nqo1, Srxn1, Prdx1, Als2, Atf1, Sod1, and Park7 were predicted. This work shows the promise of network inference algorithms operating on high-throughput gene expression data in identifying transcriptional regulatory and other signaling relationships implicated in mammalian disease