Chemically modified dsRNA induces RNAi effects in insects in vitro and in vivo: a potential new tool for improving RNA-based plant protection

Global agriculture loses over $100 billion of produce annually to crop pests such as insects. Many of these crop pests either are not currently controlled by artificial means or have developed resistance against chemical pesticides. Long dsRNAs are capable of inducing RNAi in insects and are emerging as novel, highly selective alternatives for sustainable insect management strategies. However, there are significant challenges associated with RNAi efficacy in insects. In this study, we synthesized a range of chemically modified long dsRNAs in an approach to improve nuclease resistance and RNAi efficacy in insects. Our results showed that dsRNAs containing phosphorothioate modifications demonstrated increased resistance to southern green stink bug saliva nucleases. Phosphorothioate-modified and 2′-fluoro-modified dsRNA also demonstrated increased resistance to degradation by soil nucleases and increased RNAi efficacy in Drosophila melanogaster cell cultures. In live insects, we found chemically modified long dsRNAs successfully resulted in mortality in both stink bug and corn rootworm. These results provide further mechanistic insight into the dependence of RNAi efficacy on nucleotide modifications in the sense or antisense strand of the dsRNA in insects and demonstrate for the first time that RNAi can successfully be triggered by chemically modified long dsRNAs in insect cells or live insects

Selective Inhibitors of Protozoan Protein N-myristoyltransferases as Starting Points for Tropical Disease Medicinal Chemistry Programs

Inhibition of N-myristoyltransferase has been validated pre-clinically as a target for the treatment of fungal and trypanosome infections, using species-specific inhibitors. In order to identify inhibitors of protozoan NMTs, we chose to screen a diverse subset of the Pfizer corporate collection against Plasmodium falciparum and Leishmania donovani NMTs. Primary screening hits against either enzyme were tested for selectivity over both human NMT isoforms (Hs1 and Hs2) and for broad-spectrum anti-protozoan activity against the NMT from Trypanosoma brucei. Analysis of the screening results has shown that structure-activity relationships (SAR) for Leishmania NMT are divergent from all other NMTs tested, a finding not predicted by sequence similarity calculations, resulting in the identification of four novel series of Leishmania-selective NMT inhibitors. We found a strong overlap between the SARs for Plasmodium NMT and both human NMTs, suggesting that achieving an appropriate selectivity profile will be more challenging. However, we did discover two novel series with selectivity for Plasmodium NMT over the other NMT orthologues in this study, and an additional two structurally distinct series with selectivity over Leishmania NMT. We believe that release of results from this study into the public domain will accelerate the discovery of NMT inhibitors to treat malaria and leishmaniasis. Our screening initiative is another example of how a tripartite partnership involving pharmaceutical industries, academic institutions and governmental/non-governmental organisations such as Medical Research Council and Wellcome Trust can stimulate research for neglected diseases

A seasonally varying biotope at Signy Island, Antarctic: implications for meiofaunal structure

Under the typical high-latitude conditions of temperature, productivity and settlement of chlorophyll and phaeophytin to the sediment, many benthic organisms in the Antarctic show strong seasonal variation. Although meiofauna comprise an important component of the southpolar benthic ecosystem, our knowledge of them is limited. The metazoan meiofauna and the surrounding sediments were studied fortnightly for 18 mo in a shallow bay at Signy Island (Factory Cove, South Orkneys, Antarctica) to test whether and how the temporal variability of the environment influenced meiobenthos dynamics. By examination of the distribution of the abundance and biomass of the total community, the density of higher taxonomic groups, and of individual dominant nematode genera and feeding categories, we assessed changes in faunal structure. Short-term variations were often effective, and several correlations were observed between temperature and food availability (chlorophyll and its derivatives and bulk organic matter, C and N, in both sediment and water column). However, complex temporal patterns characterised the otherwise fairly predictable seasonal variations of the Antarctic ecosystem. The results suggest that variations in meiobenthic population density and structure were primarily regulated by the input and availability of organic matter and less so by water temperature, which was constantly low. The virtual lack of a Œwinter stop¹ also leads to the conclusion that food was not limiting in the Antarctic coastal sediment

The metazoan meiofauna in its biogeochemical environment: the case of an Antarctic coastal sediment

The metazoan meiobenthos was investigated in an Antarctic coastal sediment (Factory Cove, Signy Island, Antarctica). The fine sands contained much higher abundances compared to major sublittoral sediments worldwide. Classified second after Narrangansett Bay (North Atlantic) they reached numbers of 13 × 106 ind m-2. The meiofauna was highly abundant in the surface layers, but densities decreased sharply below 2 cm. Vertical profiles mirrored steep gradients of microbiota, chloropigments and organic matter and were coincident with chemical stratification. Spatial patchiness manifested especially in the surface layer. Nematodes dominated (up to 90%), and Aponema, Chromctdorita, Diplolaimella, Daptonema, Microlaimus and Neochromadora constituted almost the entire community. Overall, the nematode fauna showed a strong similarity with fine sand communities elsewhere. The dominant trophic strategies were epistrarum and non-selective deposit feeding, but the applied classification for feeding guild structure of the nematodes of Factory Cove is discussed. High standing stock, low diversity and shallow depth distribution may have occurred because of the high nutritive (chlorophyll exceeded lOOOmgm-2 and constituted almost 50% of the organic pool) and reductive character of the benthic environment. These observations must have originated from the substantial input of fresh organic matter from phytoplankton and microphytobenthic production, typical for an Antarctic coastal ecosystem during the austral summer