Microsatellite loci for the white-dotted mosquito (Culex restuans), a principal vector of West Nile virus in North America

We characterized the first microsatellite loci in the white-dotted mosquito, Culex restuans, a critical early spring West Nile virus vector. An enrichment protocol yielded 960 positive clones of which we sequenced 300. We designed primers to amplify 29 unique di-, tri- and tetranucleotide microsatellites and chose 17 that amplified consistently across populations and were polymorphic. We developed three multiplex primer combinations for all 17 loci. A survey of 44 individuals revealed two to 20 alleles across loci, and expected heterozygosity ranging from 0.17 to 0.89. These markers will allow examination of the life history of this mysterious early season encephalitis vector.     

Mutagenesis of D80-82 and G83 residues in West Nile Virus NS2B: Effects on NS2B-NS3 activity and viral replication

Flaviviral NS2B is a required cofactor for NS3 serine protease activity and plays an important role in promoting functional NS2B-NS3 protease configuration and maintaining critical interactions with protease catalysis substrates. The residues D⁸⁰DDG in West Nile virus (WNV) NS2B are important for protease activity. To investigate the effects of D⁸⁰DDG in NS2B on protease activity and viral replication, the negatively charged region D⁸⁰DD and the conserved residue G83 of NS2B were mutated (D⁸⁰DD/E⁸⁰EE, D⁸⁰DD/K⁸⁰KK, D⁸⁰DD/A⁸⁰AA, G83F, G83S, G83D, G83K, and G83A), and NS3 D75A was designated as the negative control. The effects of the mutations on NS2B-NS3 activity, viral translation, and viral RNA replication were analyzed using kinetic analysis of site-directed enzymes and a transient replicon assay. All substitutions resulted in significantly decreased enzyme activity and blocked RNA replication. The negative charge of D⁸⁰DD is not important for maintaining NS2B function, but side chain changes in G83 have dramatic effects on protease activity and RNA replication. These results demonstrate that NS2B is important for viral replication and that D⁸⁰DD and G83 substitutions prevent replication; they will be useful for understanding the relationship between NS2B and NS3.     

Effects of NS2B-NS3 protease and furin inhibition on West Nile and Dengue virus replication

West Nile virus (WNV) and Dengue virus (DENV) replication depends on the viral NS2B-NS3 protease and the host enzyme furin, which emerged as potential drug targets. Modification of our previously described WNV protease inhibitors by basic phenylalanine analogs provided compounds with reduced potency against the WNV and DENV protease. In a second series, their decarboxylated P1-trans-(4-guanidino)cyclohexylamide was replaced by an arginyl-amide moiety. Compound 4-(guanidinomethyl)-phenylacetyl-Lys-Lys-Arg-NH₂ inhibits the NS2B-NS3 protease of WNV with an inhibition constant of 0.11?µM. Due to the similarity in substrate specificity, we have also tested the potency of our previously described multibasic furin inhibitors. Their further modification provided chimeric inhibitors with additional potency against the WNV and DENV proteases. A strong inhibition of WNV and DENV replication in cell culture was observed for the specific furin inhibitors, which reduced virus titers up to 10,000-fold. These studies reveal that potent inhibitors of furin can block the replication of DENV and WNV.     

Hominin occupations at the Dmanisi site, Georgia, Southern Caucasus: raw materials and technical behaviours of Europe's first hominins

Dmanisi is the oldest site outside of Africa that records unquestioned hominin occupations as well as the dispersal of hominins in Europe and Asia. The site has yielded large numbers of artefacts from several periods of hominin occupation. This analysis of Dmanisi stone tool technology includes a review of all the pieces recovered during the last 15 years of excavations. This lithic assemblage gives insights into the hominin behaviour at 1.7-1.8 Ma in Eurasia. Dmanisi hominins exploited local rocks derived from either nearby riverbeds or outcrops, and petrographic study provides data on patterns of stone procurement. Recent geological surveys and technological studies of the artefacts illustrate the roles of hominins in composing the assemblage. Dmanisi hominins selected two types of blanks, including cobbles and angular blocks, of basalt, andesite, and tuffs. Many complete cobbles, pebbles, and rolled blocks in basalt were unmodified, and geological analyses and surveys indicate that hominins brought manuports back to the site, suggesting a complex procurement strategy. Cores, flakes and debris show that all stages of flaking activity took place at the site. Numerous unifacial cores suggest that knapping was not very elaborate. Centripetal knapping is observed on some flake-cores. Knapping was influenced by the blank shape and natural angles. Most flaked objects were either cores or chopper-cores. Flakes predominate while flake tools are rare. The Dmanisi lithic assemblage is comparable to Oldowan sites in Africa in terms of reduction sequence, organisation of the removals, platform types, and the lack of retouched flakes. Dmanisi artefacts and may have been produced by the original hominins in Europe and Asia.     

Diatoms of Dindifelou fall (upper Basin of the Gambia River,Senegal): floristic inventory

The Dindifelou fall, located in south‐eastern Senegal, receives its water from discharging springs located in Dande plate‐land. These waters form, at the foot of the cliff, a pond that drains into the Thiokoye, a tributary of the Gambia River. The water quality of the pond and the stream is very good based on physicochemical analyses. Around the pond region, a microclimate characterized by a higher humidity and a lower temperature compared with the surrounding area is present. Diatoms study is carried out in different habitat types associated with this fall. The samples were taken by scraping the surface of rocks and rinsing fern leaves with sparkling water. Dande plate‐land is made of Upper Proterozoic sandstones whose gravity accumulations litter the bottom of the pond and the stream. This study allowed us to inventory 61 species and varieties of diatoms belonging to 28 genera. The most represented genera are Achnanthidium (ten species), Eunotia and Pinnularia (eight species each). Twelve species and varieties are reported for the first time in Senegambia including five of the genera Achnanthidium. The microflora is dominated by Achnantidium minutissimum and Achnanthidium catenatum, followed by Ulnaria ulna and various species of the genera Eunotia and Brachysira. Aerial habitats are more diversified here than the epilithic samples.     

Structural and functional characterization of an anti‐West Nile virus monoclonal antibody and its single‐chain variant produced in glycoengineered plants

Previously, our group engineered a plant‐derived monoclonal antibody (MAb pE16) that efficiently treated West Nile virus (WNV) infection in mice. In this study, we developed a pE16 variant consisting of a single‐chain variable fragment (scFv) fused to the heavy chain constant domains (C_H) of human IgG (pE16scFv‐C_H). pE16 and pE16scFv‐C_H were expressed and assembled efficiently in Nicotiana benthamiana ?XF plants, a glycosylation mutant lacking plant‐specific N‐glycan residues. Glycan analysis revealed that ?XF plant‐derived pE16scFv‐C_H (?XFpE16scFv‐C_H) and pE16 (?XFpE16) both displayed a mammalian glycosylation profile. ?XFpE16 and ?XFpE16scFv‐C_H demonstrated equivalent antigen‐binding affinity and kinetics, and slightly enhanced neutralization of WNV in vitro compared with the parent mammalian cell‐produced E16 (mE16). A single dose of ?XFpE16 or ?XFpE16scFv‐C_H protected mice against WNV‐induced mortality even 4 days after infection at equivalent rates as mE16. This study provides a detailed tandem comparison of the expression, structure and function of a therapeutic MAb and its single‐chain variant produced in glycoengineered plants. Moreover, it demonstrates the development of anti‐WNV MAb therapeutic variants that are equivalent in efficacy to pE16, simpler to produce, and likely safer to use as therapeutics due to their mammalian N‐glycosylation. This platform may lead to a more robust and cost‐effective production of antibody‐based therapeutics against WNV infection and other infectious, inflammatory or neoplastic diseases.     

The Contribution of Paraserianthes (Albizia) falcataria to Sustainable Swidden Management Practices among The Baduy of West Java

There is a prohibition on the use of modern external inputs, such as chemical fertilizers, in Baduy swidden farming. Ordinarily, the consequence of this in a situation of acute forest pressure would be a decrease in fallow times, and an inevitable depletion of soil fertility. The Baduy (particularly Outer Baduy) have several ways of alleviating this problem, one of which is the introduction of Paraserianthes (Albizia) falcataria. By alternating this commercially valuable perennial leguminous tree with rice, soil fertility is maintained and the socio-economic position of the Baduy improved. As a result, swidden farming, which is considered by the Baduy to be central to their cultural identity, continues to be viable, despite increasing population density and the continuing depletion of mature forest.     

Herpetofaunal communities at Muni Lagoon in Ghana

A herpetofaunal survey of Muni-Pomadze Lagoon, during the main rainy season (May–June), recorded a total of 32 species (13 amphibians and 19 reptiles). Three species are the first records for coastal habitats in Ghana: Kinixys homeana, Calabaria reinhardti, and Bothrophthalmus lineatus. None of the surveyed species are restricted to Ghana. The most diverse herpetofaunal community occurs in grassland thicket, with 26 species (81% of the sampled herpetofauna). Freshwater ponds include 12 species, while only three species (all amphibians) occur in brackish lagoons. Herpetofaunal conservation efforts at these sites should focus on protecting and monitoring grassland coastal thicket and freshwater ponds. Currently, freshwater pond habitats appear to be stable, but coastal thicket is being lost to provide new agricultural land at Muni. Hyperolius reedfrogs, pythons (Python regius) and monitor lizards (Varanus exanthematicus and V. niloticus) appear to offer the greatest potential for undertaking long-term monitoring of the coastal lagoon herpetofauna, because they can be more readily surveyed, and are likely to be sensitive to a broad range of environmental stressors.     

Variability in host plant resistance of Sorghum to Striga Hermonthica infestation in West Africa

Fourteen elite sorghum lines were evaluated for their resistance to Striga hermonthica at three locations in Nigeria and Mali. Results showed that many of the lines especially MALISOR 84-1, SAMSORG 41, 97-SB-F5DT-64 (Keninkédié) and the check SRN 39 remained resistant to Striga in all locations with low emerged Striga counts, while SAMSORG 14 had the highest Striga infestation in all locations. Considerable variation in reaction to Striga infestation was observed on Séguètana, 97-SB-F5DT-63 (Wasa), 97-SB-F5DT-65, CMDT 38, CMDT 39 and CMDT 45 which were susceptible to Striga at Samaru, Nigeria but were resistant to Striga at both locations in Mali. Based on low Striga resistance and high grain yield, lines MALISOR 84-1, SAMSORG 41, 97-SB-F5DT-64, 97-SB-F5DT-65, CMDT 39 and SAMSORT 14 have been nominated for wider evaluation across more West African countries.