Solid-phase synthesis of terminal oligonucleotide-phosphoramidate conjugates

A novel phosphoramidite, N,N-diisopropylamino-2-cyanoethyl-9-anthracenemethyl phosphoramidite 1, was prepared and coupled with the terminal 5'-hydroxyl of support-bound T10 and the putative phosphite triester intermediate was subsequently reacted with iodine in the presence of either water or a series of primary and secondary amines. The reactivity of 1 compared to a previously reported benzyl phosphoramidite 2 was also investigated: oxidation of the product of coupling 2 with CPG-T10-5'OH under aqueous conditions resulted in greater than 30% of the benzyl moiety being retained. In contrast, essentially complete loss of the 9-anthracenemethyl group was observed using 1 under the same conditions. Oligonucleotides modified with a terminal phosphate monoester, lipophilic, fluorescent or cationic groups were thus prepared.

Pollinators of an endemic and endangered species, Mammillaria gaumeri (Cactaceae), in its natural habitat (coastal dune) and in a botanical garden.

Mammillaria gaumeri (Britton & Ross) Orcutt (Cactaceae), an endemic plant of the Yucatan Peninsula, is included by the Mexican government in the list of species that require special protection. Its natural habitat is now restricted to fragmented areas and protection programs involve botanical gardens in growing individuals rescued from disturbed areas. Little information is available on the reproductive characteristics of this species and nothing is known of its pollinators. We investigated the visitors of M. gaumeri flowers, collecting and observing bee species in its natural habitat (i.e., coastal dune) and in a botanical garden, where coastal dune vegetation had been created. Observations were made on plants whose density was artificially increased by grouping flowering individuals. At each site, we: 1) collected insects visiting the flowers; 2) recorded number of visits; and 3) video-recorded bee movements on the flowers. As expected, the number of bee species and visitation frequency were higher at the botanical garden than at the coastal dune. After landing on a flower, bees either inspected the anthers or dived among them. These behaviors, carried out by all observed species, seemed related to the state of the anthers (full or empty of pollen) and stigma lobes (opened or closed). Specifically, visits lasted longer when anthers were full of pollen and stigma lobes were opened. The same bee species recorded on the dune were also recorded at the botanical garden, suggesting that the artificial dune at the botanical garden offered suitable conditions for the natural pollinators of this endangered cactus.

Satellite flies (Leucophora personata, Diptera : Anthomyiidae) and other dipteran parasites of the communal bee Andrena agilissima (Hymenoptera : Andrenidae) on the island of Elba, Italy

Solitary and presocial aculueate Hymenoptera are parasitized by a range of dipteran species in the families Axithomyiidae, Bombyliidae, Conopidae, Phoridae, and Sarcophagidae that are likely to impact on their hosts. We undertook a study over several years of a univoltine and communal bee, Andrena agilissima, and its main dipteran parasites, in particular the satellite fly Leucophora personata (Diptera: Anthomyiidae). Behavioural and ecological data were collected from one nesting aggregation of the host bee on the island of Elba, Italy, from 1993 to 2003, and from a foraging site of the bee, ca 5 km from the nesting aggregation. Other Diptera associated with A. agilissmia at the field site were the bee fly Bombylius fimbriatus (Bombyliidae), the conopid fly Zodion cinereum (Conopidae), and the scuttle fly Megaselia andrenae (Phoridae). The phenology of the Diptera broadly overlapped with that of their host across the season of activity (end of April and all of May). Diurnal activity patterns differed slightly; L. personata in particular was active at the host's nesting site before A. agilissima. Female satellite flies also showed a range of behaviours in gaining entry to a host nest. We summarize published data on this and other Leucophora species that parasitize Andrena host bees. Host bees returning to their nests occasionally undertook zig-zag flight manoeuvres if followed by a satellite fly that were generally successful in evading the fly. Satellite flies that entered a nest, presumably to oviposit, were less likely to remain therein if another host bee entered the same nest, suggesting that one advantage to communal nesting for this host is a reduction in brood cell parasitism by L. personata. We provide the first clear evidence for parasitism by a Zodion of any Andrena host. Both L. personata and M. andrenae concentrated their parasitic activities in the zone of the host nesting aggregation with highest nest densities. Three of the Diptera, L. personata, B. fimbriatus, and Z. cinereum, seemed to have extremely low rates of parasitism whilst that of M. andrenae appeared low. Though they have refined parasitic behaviour that allows them to gain entry into host nests (L. personata, B. fimbriatus, and M. andrenae) or to parasitize adults (Z. cinercum), these parasites seem not to impact upon the dynamics of the host A. agilissima at the nesting aggregation, and the host possesses traits to reduce parasitism.

Antibiofilm activities of 1-alkyl-3-methylimidazolium chloride ionic liquids

Microbial biofilms are ubiquitous in nature and represent the predominant mode of growth of microorganisms. A general characteristic of biofilm communities is that they tend to exhibit significant tolerance to antimicrobial challenge compared with planktonic bacteria of the same species The antibiofilm activity of a series of 1-alkyl-3-methylimidazolium chloride ionic liquids has been evaluated against a panel of clinically significant microbial pathogens, including MRSA. A comparison of antimicrobial activity against planktonic bacteria and established biofilms is presented. In general, these ionic liquids possess potent, broad spectrum antibiofilm activity.

Antimicrobial and antibiofilm activities of 1-alkylquinolinium bromide ionic liquids

Quinoline derivatives are known to possess a range of bioactive and medicinal activities, which have been exploited in the design of antibacterial, antifungal and antimalarial compounds. In this study, we report on the microbiological toxicity of a series of 1-alkylquinolinium bromides against a range of clinically relevant microorganisms, in both planktonic and sessile (biofilm) cultures. A comparison of antimicrobial activity against planktonic bacteria and established biofilms is presented. In general, 1-alkylquinolinium ionic liquids possess excellent, broad spectrum antimicrobial activity against microorganisms grown in both the planktonic and sessile, or biofilm, mode of growth. Importantly, these compounds are potent against Gram positive and Gram negative bacteria, as well as fungi, with a clear dependency on length of the alkyl substituent for activity, with compounds containing twelve and fourteen carbons in the alkyl group exhibiting highest antimicrobial and antibiofilm activity. © 2010 The Royal Society of Chemistry.

Cytotoxicity of 1-alkylquinolinium bromide ionic liquids in murine fibroblast NIH 3T3 cells

Minimal toxicity data are available for 1-alkylquinolinium bromide ionic liquids. Here, their toxicity to NIH 3T3 murine fibroblast cells, of relevance to their potential antimicrobial application, is presented. Toxicity data, presented by time-point analysis with a particular focus on the immediate toxicity upon short term cellular exposure, indicate a link between the length of the alkyl chain substituent and resultant biological toxicity. 1-Tetradecylquinolinium bromide was found to exhibit cellular toxicity comparable to benzalkonium chloride over all time points tested. By comparison, 1-octylquinolinium bromide initially exerted significantly lower cytotoxicity at one hour; however, toxicity was found to have a cumulative effect over time-course analysis up to three days. This illustrates that alkyl chain components may govern not only overall toxicity, but also the rate of toxicity. Fluorescence microscopy was utilised to examine destabilisation of the plasma membrane by 1 tetradecylquinolinium bromide and benzalkonium chloride after one hour, with membrane destabilisation not observed for 1-octylquinolinium bromide, or the base constituent quinoline.