Structural and biochemical analysis of the death domain complex formed at the Fas receptor.

Fas (CD95) is a member of the death receptor superfamily of proteins that are involved in the initiation of apoptosis as induced by the binding of extracellular ligands. At present, little is known about the precise mechanism by which the signal initiated by the interaction between Fas ligand (FasL) and Fas is transduced across the cell membrane to start the apoptotic signalling cascade. The first step in this pathway is the recruitment of the Fas-associated death domain protein (FADD) to the cytoplasmic death domain of Fas via a homotypic protein/protein interaction. This binding event occurs after receptor ligation apparently without any post translational modification such as phosphorylation. In order to better understand this event we have investigated the interaction between the death domains of the human Fas and FADD proteins both in vitro and in a cellular context. The reported interaction between the Fas and FADD death domains (Fas-DD and FADD-DD) was recapitulated using the yeast 2-hybrid assay. Recombinant proteins were then produced for NMR spectroscopy experiments. FADD-DD is highly expressed, and easy to isolate soluble at physiological pH. This domain is readily expressed as a histidine tagged domain. Fas death domain expresses at low levels and produces soluble aggregates when concentrated at a pH above 4. However, it was found that using a Gbl fusion protein to express Fas-DD overcomes these problems and allowed the production of Fas-DD for NMR experiments. NMR titration experiments showed that when these two proteins interact a large, soluble complex is formed. This may be significant in relation to the increasing evidence for the importance of Fas-receptor clustering in its signalling. Mutational analysis of the Fas death domain was also carried out. Here, various previously described as well as several novel point mutations were made on the surface of the Fas death domain to investigate their effect on FADD-DD binding. These mutations were assayed using yeast 2-hybrid methods, NMR analysis and in a cell based assay. In the cell based assay wild type and mutant Fas receptors were overexpressed in a human cell line with no endogenous surface Fas expression. These cells were then assayed for their ability to undergo FasL-induced apoptosis. It was found that residues from many surface regions of Fas-DD are crucial for the FADD-DD interaction. This observation has potentially important implications for the nature of the organisation of the death domains in the death inducing signalling complex (DISC) formed at the Fas receptor

Artemisinin-based combination therapy does not measurably reduce human infectiousness to vectors in a setting of intense malaria transmission

<p>Background: Artemisinin-based combination therapy (ACT) for treating malaria has activity against immature gametocytes. In theory, this property may complement the effect of terminating otherwise lengthy malaria infections and reducing the parasite reservoir in the human population that can infect vector mosquitoes. However, this has never been verified at a population level in a setting with intense transmission, where chronically infectious asymptomatic carriers are common and cured patients are rapidly and repeatedly re-infected.</p> <p>Methods: From 2001 to 2004, malaria vector densities were monitored using light traps in three Tanzanian districts. Mosquitoes were dissected to determine parous and oocyst rates. Plasmodium falciparum sporozoite rates were determined by ELISA. Sulphadoxinepyrimethamine(SP) monotherapy was used for treatment of uncomplicated malaria in the contiguous districts of Kilombero and Ulanga throughout this period. In Rufiji district, the standard drug was changed to artesunate co-administered with SP (AS + SP) in March 2003. The effects of this change in case management on malaria parasite infection in the vectors were analysed.</p> <p>Results: Plasmodium falciparum entomological inoculation rates exceeded 300 infective bites per person per year at both sites over the whole period. The introduction of AS + SP in Rufiji was associated with increased oocyst prevalence (OR [95%CI] = 3.9 [2.9-5.3], p < 0.001), but had no consistent effect on sporozoite prevalence (OR [95%CI] = 0.9 [0.7-1.2], p = 0.5). The estimated infectiousness of the human population in Rufiji was very low prior to the change in drug policy. Emergence rates and parous rates of the vectors varied substantially throughout the study period, which affected estimates of infectiousness. The latter consequently cannot be explained by the change in drug policy.</p> <p>Conclusions: In high perennial transmission settings, only a small proportion of infections in humans are symptomatic or treated, so case management with ACT may have little impact on overall infectiousness of the human population. Variations in infection levels in vectors largely depend on the age distribution of the mosquito population. Benefits of ACT in suppressingtransmission are more likely to be evident where transmission is already low or effective vector control is widely implemented.</p&gt

Emergence of behavioural avoidance strategies of malaria vectors in areas of high LLIN coverage in Tanzania

Despite significant reductions in malaria transmission across Africa since 2000, progress is stalling. This has been attributed to the development of insecticide resistance and behavioural adaptations in malaria vectors. Whilst insecticide resistance has been widely investigated, there is poorer understanding of the emergence, dynamics and impact of mosquito behavioural adaptations. We conducted a longitudinal investigation of malaria vector host choice over 3 years and resting behaviour over 4 years following a mass long-lasting insecticidal nets (LLINs) distribution in Tanzania. By pairing observations of mosquito ecology with environmental monitoring, we quantified longitudinal shifts in host-choice and resting behaviour that are consistent with adaptation to evade LLINs. The density of An. funestus s.l., declined significantly through time. In tandem, An. arabiensis and An. funestus s.l. exhibited an increased rate of outdoor relative to indoor resting; with An. arabiensis reducing the proportion of blood meals taken from humans in favour of cattle. By accounting for environmental variation, this study detected clear evidence of intra-specific shifts in mosquito behaviour that could be obscured in shorter-term or temporally-coarse surveys. This highlights the importance of mosquito behavioural adaptations to vector control, and the value of longer-term behavioural studies

The first long-read nuclear genome assembly of Oryza australiensis, a wild rice from northern Australia

Oryza australiensis is a wild rice native to monsoonal northern Australia. The International Oryza Map Alignment Project emphasises its significance as the sole representative of the EE genome clade. Assembly of the O. australiensis genome has previously been challenging due to its high Long Terminal Repeat (LTR) retrotransposon (RT) content. Oxford Nanopore long reads were combined with Illumina short reads to generate a high-quality ~ 858 Mbp genome assembly within 850 contigs with 46× long read coverage. Reference-guided scaffolding increased genome contiguity, placing 88.2% of contigs into 12 pseudomolecules. After alignment to the Oryza sativa cv. Nipponbare genome, we observed several structural variations. PacBio Iso-Seq data were generated for five distinct tissues to improve the functional annotation of 34,587 protein-coding genes and 42,329 transcripts. We also report SNV numbers for three additional O. australiensis genotypes based on Illumina re-sequencing. Although genetic similarity reflected geographical separation, the density of SNVs also correlated with our previous report on variations in salinity tolerance. This genome re-confirms the genetic remoteness of the O. australiensis lineage within the O. officinalis genome complex. Assembly of a high-quality genome for O. australiensis provides an important resource for the discovery of critical genes involved in development and stress tolerance.Aaron L. Phillips, Scott Ferguson, Nathan S. Watson, Haigh, Ashley W. Jones, Justin O. Borevitz, Rachel A. Burton, Brian J. Atwel

Nutrigenomics, the microbiome, and gene-environment interactions: New directions in cardiovascular disease research, prevention, and treatment

Cardiometabolic diseases are the leading cause of death worldwide and are strongly linked to both genetic and nutritional factors. The field of nutrigenomics encompasses multiple approaches aimed at understanding the effects of diet on health or disease development, including nutrigenetic studies investigating the relationship between genetic variants and diet in modulating cardiometabolic risk, as well as the effects of dietary components on multiple "omic" measures, including transcriptomics, metabolomics, proteomics, lipidomics, epigenetic modifications, and the microbiome. Here, we describe the current state of the field of nutrigenomics with respect to cardiometabolic disease research and outline a direction for the integration of multiple omics techniques in future nutrigenomic studies aimed at understanding mechanisms and developing new therapeutic options for cardiometabolic disease treatment and prevention

A pleurocidin analogue with greater conformational flexibility, enhanced antimicrobial potency and in vivo therapeutic efficacy.

Antimicrobial peptides (AMPs) are a potential alternative to classical antibiotics that are yet to achieve a therapeutic breakthrough for treatment of systemic infections. The antibacterial potency of pleurocidin, an AMP from Winter Flounder, is linked to its ability to cross bacterial plasma membranes and seek intracellular targets while also causing membrane damage. Here we describe modification strategies that generate pleurocidin analogues with substantially improved, broad spectrum, antibacterial properties, which are effective in murine models of bacterial lung infection. Increasing peptide-lipid intermolecular hydrogen bonding capabilities enhances conformational flexibility, associated with membrane translocation, but also membrane damage and potency, most notably against Gram-positive bacteria. This negates their ability to metabolically adapt to the AMP threat. An analogue comprising D-amino acids was well tolerated at an intravenous dose of 15 mg/kg and similarly effective as vancomycin in reducing EMRSA-15 lung CFU. This highlights the therapeutic potential of systemically delivered, bactericidal AMPs

Realising the health and wellbeing of adolescents

Adolescence is a critical stage of life characterised by rapid biological, emotional, and social development. It is during this time that every person develops the capabilities required for a productive, healthy, and satisfying life. In order to make a healthy transition into adulthood, adolescents need to have access to health education, including education on sexuality1; quality health services, including sexual and reproductive; and a supportive environment both at home and in communities and countries.The global community increasingly recognises these vital needs of adolescents, and there is an emerging consensus that investing intensively in adolescents’ health and development is not only key to improving their survival and wellbeing but critical for the success of the post-2015 development agenda.2 The suggested inclusion of adolescent health in the United Nations secretary general’s Global Strategy for Women’s and Children’s Health is an expression of this growing awareness and represents an unprecedented opportunity to place adolescents on the political map beyond 2015. Ensuring that every adolescent has the knowledge, skills, and opportunities for a healthy, productive life and enjoyment of all human rights3 is essential for achieving improved health, social justice, gender equality, and other development goals.We argue that the priority in the revised Every Women Every Child Global Strategy needs to be giving adolescents a voice, expanding their choices and control over their bodies, and enabling them to develop the capabilities required for a productive, healthy, and satisfying life. We call for a global, participatory movement to improve the health of the world’s adolescents as part of a broader agenda to improve their wellbeing and uphold their rights