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With emerging resistance to frontline treatments, it is vital that new antimalarial drugs are identified to target Plasmodium falciparum. We have recently described a compound, MMV020291, as a specific inhibitor of red blood cell (RBC) invasion, and have generated analogues with improved potency. Here, we generated resistance to MMV020291 and performed whole genome sequencing of 3 MMV020291-resistant populations. This revealed 3 nonsynonymous single nucleotide polymorphisms in 2 genes; 2 in profilin (N154Y, K124N) and a third one in actin-1 (M356L). Using CRISPR-Cas9, we engineered these mutations into wild-type parasites, which rendered them resistant to MMV020291. We demonstrate that MMV020291 reduces actin polymerisation that is required by the merozoite stage parasites to invade RBCs. Additionally, the series inhibits the actin-1-dependent process of apicoplast segregation, leading to a delayed death phenotype. In vitro cosedimentation experiments using recombinant P. falciparum proteins indicate that potent MMV020291 analogues disrupt the formation of filamentous actin in the presence of profilin. Altogether, this study identifies the first compound series interfering with the actin-1/profilin interaction in P. falciparum and paves the way for future antimalarial development against the highly dynamic process of actin polymerisation.

The resource valuation and optimisation model (RVOM) is a partial differential equation-based real options software package, which helps mine owners optimally plan operations, understand project risks, and make defensible valuations. This is achieved in the presence of both financial and physical uncertainty, as well as processing capacity constraints. The RVOM can be applied to any multi-ore mine where the block-order of extraction has already been planned using existing software tools. The RVOM can also take account of multiple commodities within a single mine, and multiple forms of price behaviour. The three key outputs from the RVOM are: valuation, optimal decision and probability of decision. A decision takes account of upfront costs, and can include an unlimited number of transitions between normal operation, expanded operation, care and maintenance and abandonment (and variants thereof). Use of the RVOM in a gold mine case-study is discussed, demonstrating its broad applicability, the added value it can create and its ease of use., The resource valuation and optimisation model (RVOM) is a partial differential equation-based real options software package, which helps mine owners optimally plan operations, understand project risks, and make defensible valuations. This is achieved in the presence of both financial and physical uncertainty, as well as processing capacity constraints. The RVOM can be applied to any multi-ore mine where the block-order of extraction has already been planned using existing software tools. The RVOM can also take account of multiple commodities within a single mine, and multiple forms of price behaviour. The three key outputs from the RVOM are: valuation, optimal decision and probability of decision. A decision takes account of upfront costs, and can include an unlimited number of transitions between normal operation, expanded operation, care and maintenance and abandonment (and variants thereof). Use of the RVOM in a gold mine case-study is discussed, demonstrating its broad applicability, the added value it can create and its ease of use.

BACKGROUND: Progressive multifocal leukoencephalopathy (PML) in natalizumab-treated MS patients is linked to JC virus (JCV) infection. JCV sequence variation and rearrangements influence viral pathogenicity and tropism. To better understand PML development, we analyzed viral DNA sequences in blood, CSF and/or urine of natalizumab-treated PML patients. METHODS: Using biofluid samples from 17 natalizumab-treated PML patients, we sequenced multiple isolates of the JCV noncoding control region (NCCR), VP1 capsid coding region, and the entire 5 kb viral genome. RESULTS: Analysis of JCV from multiple biofluids revealed that individuals were infected with a single genotype. Across our patient cohort, multiple PML-associated NCCR rearrangements and VP1 mutations were present in CSF and blood, but absent from urine-derived virus. NCCR rearrangements occurred in CSF of 100% of our cohort. VP1 mutations were observed in blood or CSF in 81% of patients. Sequencing of complete JCV genomes demonstrated that NCCR rearrangements could occur without VP1 mutations, but VP1 mutations were not observed without NCCR rearrangement. CONCLUSIONS: These data confirm that JCV in natalizumab-PML patients is similar to that observed in other PML patient groups, multiple genotypes are associated with PML, individual patients appear to be infected with a single genotype, and PML-associated mutations arise in patients during PML development.

Introduction The importance of eucalypts and reasons for tissue culture Eucalypts are Australia's most distinctive plant group. They are contained within the genus Eucalyptus which consists of over 500 named species, with more as yet unnamed (Brooker & Kleinig 1983; 1990 Chippendale 1988). The natural distribution of the genus is almost completely confined to the Australian continent and Tasmania with only two species, E. deglupta and E. urophylla, occurring naturally in other countries. Since European settlement of Australia, seeds of eucalypts have been sent to countries throughout the world and they are now commonly grown in tropical and temperate areas for timber, pulp wood, eucalyptus oil, fuelwood, charcoal and as ornamentals. Exploitation of eucalypts outside Australia was initiated by the French. During the nineteenth century, eucalypts were planted in Europe and North America, and European imperial governments introduced them to colonies in South America, Africa and Asia. The presence of eucalypts in some of these countries is now so familiar to the native peoples that many consider them to be indigenous (Zacharin 1978). Although eucalypts in early plantations often grew very quickly the wood was sometimes of poor quality due to wood splitting and distortion (Clarke 1957; Penfold & Willis 1961; Pryor 1976). In many cases this was because the species chosen were inappropriate for local climatic and edaphic conditions (Evans 1980; Durand-Cresswell et al. 1982), the trees had been planted for the wrong purposes (Penfold & Willis 1961; Pryor 1976), or given incorrect fertilisers (Savory 1962; Stone 1968). The poor quality of the wood led to a slump in enthusiasm for growing eucalypts until about 1945 when world demand for pulpwood started to increase (Pryor 1976). Today the major uses of eucalypt wood are for fuelwood and pulpwood. There has been a 150 fold increase in pulpwood production from eucalypts since the early 1960s (Molleda 1984). They are now the mos