Adequate clinical and parasitologic cure by artemisinin combination therapies relies on the artemisinin component and the partner drug. Polymorphisms in the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and P. falciparum multidrug resistance 1 (pfmdr1) genes are associated with decreased sensitivity to amodiaquine and lumefantrine, but effects of these polymorphisms on therapeutic responses to artesunate-amodiaquine (ASAQ) and artemether-lumefantrine (AL) have not been clearly defined. Individual patient data from 31 clinical trials were harmonized and pooled by using standardized methods from the WorldWide Antimalarial Resistance Network. Data for more than 7,000 patients were analyzed to assess relationships between parasite polymorphisms in pfcrt and pfmdr1 and clinically relevant outcomes after treatment with AL or ASAQ. Presence of the pfmdr1 gene N86 (adjusted hazards ratio = 4.74, 95% confidence interval = 2.29 – 9.78, P < 0.001) and increased pfmdr1 copy number (adjusted hazards ratio = 6.52, 95% confidence interval = 2.36–17.97, P < 0.001) were significant independent risk factors for recrudescence in patients treated with AL. AL and ASAQ exerted opposing selective effects on single-nucleotide polymorphisms in pfcrt and pfmdr1. Monitoring selection and responding to emerging signs of drug resistance are critical tools for preserving efficacy of artemisinin combination therapies; determination of the prevalence of at least pfcrt K76T and pfmdr1 N86Y should now be routine.

The human chemokine receptors CCR5 and CXCR4 have emerged as the predominant cofactors, along with CD4, for cellular entry of HIV-1 in vivo whereas the contribution of other chemokine receptors to HIV disease has not been yet determined. CCR5-specific (R5) viruses predominate during primary HIV-1 infection whereas viruses with specificity for CXCR4 (R5͞X4 or X4 viruses) often emerge in late stages of HIV disease. The evolution of X4 viruses is associated with a rapid decline in CD4؉ T cells, although a causative relationship between viral tropism and CD4؉ T cell depletion has not yet been proven. To rigorously test this relationship, we assessed CD4؉ T cell depletion in suspensions of human peripheral blood mononuclear cells and in explants of human lymphoid tissue on exposure to paired viruses that are genetically identical (isogenic) except for select envelope determinants specifying reciprocal tropism for CXCR4 or CCR5. In both systems, X4 HIV-1 massively depleted CD4؉ lymphocytes whereas matched R5 viruses depleted such cells only mildly despite comparable viral replication kinetics. These findings demonstrate that the coreceptor specificities of HIV-1 are a causal factor in CD4؉ T cell depletion ex vivo and strongly support the hypothesis that the evolution of viral envelope leading to usage of CXCR4 in vivo accelerates loss of CD4؉ T cells, causing immunodeficiency.

Although infection with vaccinia virus (VV) is known to affect the cytoskeleton, it is not known how this affects the cellular architecture or whether the attenuated modified VV ankara (MVA) behaves similar to wild-type VV (wtVV). In the present study, we therefore compared effects of wtVV and MVA infection on the cellular architecture. WtVV-infection induces cell rounding early in infection, which coincides with the retraction of microtubules (MTs) and intermediate filaments from the cellular periphery, whereas mitochondria and late endosomes cluster around the nucleus. Nocodazole treatment demonstrates that cell rounding and organelle clustering require intact MTs. At the onset of virus assembly late in infection, cells reflatten, a process that coincides with the regrowth of MTs into the cellular periphery. We find that the actin network undergoes several rearrangements that occur sequentially in time and that closely follow the cell-shape changes. Unexpectedly, these actin changes are blocked or reversed upon nocodazole treatment, indicating that intact MTs are also responsible for the wtVV-induced actin rearrangements. Finally, MVA infection does not induce any of these cellular changes. Because this virus lacks a substantial number of VV genes, MVA opens up a system to search for the molecules involved in wtVV-induced cellular changes; in particular, those that may regulate actin/MT interactions.Key words: actin, cell shape, microtubules, modified vaccinia virus ankara, vaccinia virus. Vaccinia virus (VV) is the prototype of the poxviridae, a family of large DNA viruses. Its genome of about 200 kB encodes for more than 250 proteins (1) enabling VV to carry out DNA replication and transcription in the cytoplasm rather than in the cellular nucleus (1). VV is characterized by a complex cytoplasmic life cycle, directed by three classes of genes, early, intermediate and late. Following entry, the viral core is delivered into the cytoplasm, from which a defined set of early mRNAs is transcribed and extruded into the cytoplasm for translation. The early proteins are required to uncoat the viral core and to initiate cytoplasmic DNA replication. Replication sets off the transcription of intermediate and late genes, late proteins being required for virion assembly. During assembly, late in infection, two infectious forms are made; the intracellular mature virus (IMV) and the extracellular enveloped virus (EEV). A small percentage of the intracellular IMVs becomes enwrapped by a double membrane derived from the transGolgi network (TGN) or endosomes to form the intracellular enveloped virus [IEV (2,3)]. The IEV moves along microtubules (MTs) towards the plasma membrane using the plusend directed motor protein kinesin-1. Upon fusion of the outer IEV membrane with the plasma membrane, the EEV is released into the extracellular environment. The outer IEV membrane that is fused with the plasma membrane is able to polymerize actin tails, resulting in long filopodia that propel attached EEVs towards neighbouring cells, a pr...

The purpose of this study was to evaluate the cost of illness of moderateto-severe atopic asthma and/or seasonal allergic rhinitis (SAR) in Germany from the perspective of third-party payers (TPP) and patients.Five-hundred patients (276 children/adolescents) with moderate-to-severe asthma and/ or SAR were included in this cross-sectional study. Information was collected using a specific patient questionnaire and the abstraction of patient records.Overall, annual costs per patient increased with the severity of atopic asthma and if it was associated with SAR. The average annual cost of SAR was J1,089 per child/ adolescent and J1,543 per adult. Annual costs of severe asthma plus SAR increased to J7,928 per child/adolescent and to J9,287 per adult. For TPPs, the main cost drivers were medication, hospitalisation, and rehabilitation. The most significant costs for patients were household modifications. For children/adolescents, 60-78% of the expenditures were direct costs, while in adults, 58% of expenditures were indirect costs. It was also observed that patients with moderate and severe asthma used inhaled corticosteroids less frequently than recommended by treatment guidelines.In summary, the total cost for patients increases with the severity of atopic asthma and/or seasonal allergic rhinitis and indirect costs represent a large proportion of the total cost.