Plasminogen alleles influence susceptibility to invasive aspergillosis

Invasive aspergillosis (IA) is a common and life-threatening infection in immunocompromised individuals. A number of environmental and epidemiologic risk factors for developing IA have been identified. However, genetic factors that affect risk for developing IA have not been clearly identified. We report that host genetic differences influence outcome following establishment of pulmonary aspergillosis in an exogenously immune suppressed mouse model. Computational haplotype-based genetic analysis indicated that genetic variation within the biologically plausible positional candidate gene plasminogen (Plg; Gene ID 18855) correlated with murine outcome. There was a single nonsynonymous coding change (Gly110Ser) where the minor allele was found in all of the susceptible strains, but not in the resistant strains. A nonsynonymous single nucleotide polymorphism (Asp472Asn) was also identified in the human homolog (PLG; Gene ID 5340). An association study within a cohort of 236 allogeneic hematopoietic stem cell transplant (HSCT) recipients revealed that alleles at this SNP significantly affected the risk of developing IA after HSCT. Furthermore, we demonstrated that plasminogen directly binds to Aspergillus fumigatus. We propose that genetic variation within the plasminogen pathway influences the pathogenesis of this invasive fungal infection.     

Computational study of the putative active form of protein Z (PZa): sequence design and structural modeling

Although protein Z (PZ) has a domain arrangement similar to the essential coagulation proteins FVII, FIX, FX, and protein C, its serine protease (SP)-like domain is incomplete and does not exhibit proteolytic activity. We have generated a trial sequence of putative activated protein Z (PZa) by identifying amino acid mutations in the SP-like domain that might reasonably resurrect the serine protease catalytic activity of PZ. The structure of the activated form was then modeled based on the proposed sequence using homology modeling and solvent-equilibrated molecular dynamics simulations. In silico docking of inhibitors of FVIIa and FXa to the putative active site of equilibrated PZa, along with structural comparison with its homologous proteins, suggest that the designed PZa can possibly act as a serine protease.     

Seasonal patterns of carbohydrate translocation and synthesis of structural carbon components in <Emphasis Type="Italic">Typha angustifolia</Emphasis>

Quantitative studies of material budgets and resource allocation patterns of emergent plants are needed to fully understand nutrient and carbon cycling in wetlands. Whole-plant translocation patterns of nonstructural carbohydrates and synthesis of structural carbon were documented using two (shallow and deepwater) populations of Typha angustifolia in floodplain habitats of the Arakawa River, Japan. Monthly and bimonthly measurements of the concentrations of total carbon, non-structural carbohydrates, and water-soluble carbohydrates, as well as, estimates of standing stocks of aboveground (AG) and belowground (BG) biomass for both populations from 2002 to 2004 are described here. Annual patterns of carbon gain, rates of carbohydrate translocation between AG and BG organs, and rates of synthesis of structural carbon were estimated. Upward translocation supported all AG production for approximately 30 days. Afterward, the fraction of AG production supported by upward translocation decreased linearly with time, and completely diminished by Day 80 (counted from first day of growth; March 15). At Day 80, material translocation was directed downward and the percentage of downward translocation relative to AG net production increased until Day 170; there was a vigorous increase in downward translocation prior to senescence. In early summer (Day 80–110), more than half of the translocated materials were transformed into structural components, but by Day 125 only 30% was synthesized into total structural carbohydrates. Prior to senescence, there was a greater percentage of nonstructural carbohydrates in the rhizomes. Across the growing season (Day 60–153), the total amount of downward translocation was found to be proportional to AG production and the quantitative evaluation of the carbon budget shed further insight into the translocation process of this rhizomatous aquatic plants. In addition, insights on the differences between shallow and deepwater populations were gained. Especially, the deepwater population supported a higher fraction of vertical rhizomes compared to individuals sampled from the shallow water population. Handling editor: S. M. Thomaz.     

Pralidoxime in Acute Organophosphorus Insecticide Poisoning—A Randomised Controlled Trial

Background

Poisoning with organophosphorus (OP) insecticides is a major global public health problem, causing an estimated 200,000 deaths each year. Although the World Health Organization recommends use of pralidoxime, this antidote''s effectiveness remains unclear. We aimed to determine whether the addition of pralidoxime chloride to atropine and supportive care offers benefit.

Methods and Findings

We performed a double-blind randomised placebo-controlled trial of pralidoxime chloride (2 g loading dose over 20 min, followed by a constant infusion of 0.5 g/h for up to 7 d) versus saline in patients with organophosphorus insecticide self-poisoning. Mortality was the primary outcome; secondary outcomes included intubation, duration of intubation, and time to death. We measured baseline markers of exposure and pharmacodynamic markers of response to aid interpretation of clinical outcomes. Two hundred thirty-five patients were randomised to receive pralidoxime (121) or saline placebo (114). Pralidoxime produced substantial and moderate red cell acetylcholinesterase reactivation in patients poisoned by diethyl and dimethyl compounds, respectively. Mortality was nonsignificantly higher in patients receiving pralidoxime: 30/121 (24.8%) receiving pralidoxime died, compared with 18/114 (15.8%) receiving placebo (adjusted hazard ratio [HR] 1.69, 95% confidence interval [CI] 0.88–3.26, p = 0.12). Incorporating the baseline amount of acetylcholinesterase already aged and plasma OP concentration into the analysis increased the HR for patients receiving pralidoxime compared to placebo, further decreasing the likelihood that pralidoxime is beneficial. The need for intubation was similar in both groups (pralidoxime 26/121 [21.5%], placebo 24/114 [21.1%], adjusted HR 1.27 [95% CI 0.71–2.29]). To reduce confounding due to ingestion of different insecticides, we further analysed patients with confirmed chlorpyrifos or dimethoate poisoning alone, finding no evidence of benefit.

Conclusions

Despite clear reactivation of red cell acetylcholinesterase in diethyl organophosphorus pesticide poisoned patients, we found no evidence that this regimen improves survival or reduces need for intubation in patients with organophosphorus insecticide poisoning. The reason for this failure to benefit patients was not apparent. Further studies of different dose regimens or different oximes are required.

Trial Registration

Controlled-trials.com ISRCTN55264358 Please see later in the article for Editors'' Summary     

Growth and phosphorus nutrition of rice when inorganic fertiliser application is partly replaced by straw under varying moisture availability in sandy and clay soils

Background and aims

The combined effects of (1) reduced soil moisture availability, (2) reduced application of inorganic fertilisers while incorporating straw, (3) soil type, and their effects on growth, root system plasticity, phosphorus (P) nutrition of rice, and soil P dynamics are poorly known, but very important when aiming to increase the efficiency of water and P use.

Methods

Using large pots a three-factor factorial experiment was conducted with two moisture treatments (i.e. continuous flooding, and draining of top soil after flowering while subsoil was kept moist through capillary action), three fertilisation treatments; with (P1) and without (P0) applications of inorganic P fertilisers, and 25 % of inorganic fertilisers reduced while incorporating straw (5 t ha^?1), and soil type (i.e. clay and sandy soils with 15 and 9 mg P kg^?1 soil, respectively in P0). Shoot and root growth, root system plasticity, P nutrient status and soil P dynamics were measured.

Key results

Straw incorporation with reduced inorganic fertiliser application ensured a higher shoot dry weight and yield only in flooded clay soil as compared with P0 and P1, and a similar shoot dry weight and yield to P1 under drained clay soil. A positive growth response was facilitated by an increased water-use efficiency and rate of photosynthesis in shoots, and increased root system plasticity through the production of greater root length, more roots in deep soil layers, and an increased fraction of fine roots. Straw enhanced P extractability in soil. Drained soil reduced P uptake (15–45 %) and increased P-use efficiency. In addition to the re-translocation of P from senescing leaves and stems under both moisture conditions, the P concentration in green leaves under drained condition was also reduced (41–72 %).

Conclusion

Growth benefits of straw incorporation were observed in clay soil under both moisture conditions, and this was facilitated by the improved P availability, increased P uptake, and greater root system plasticity with the production of deeper and finer roots, compared with that in sandy soil, and inorganic fertiliser applications alone. As P uptake was reduced under drained soil, P re-translocation and % P allocated to panicles increased.     

The Interactions of P-Glycoprotein with Antimalarial Drugs,Including Substrate Affinity,Inhibition and Regulation

The combination of passive drug permeability, affinity for uptake and efflux transporters as well as gastrointestinal metabolism defines net drug absorption. Efflux mechanisms are often overlooked when examining the absorption phase of drug bioavailability. Knowing the affinity of antimalarials for efflux transporters such as P-glycoprotein (P-gp) may assist in the determination of drug absorption and pharmacokinetic drug interactions during oral absorption in drug combination therapies. Concurrent administration of P-gp inhibitors and P-gp substrate drugs may also result in alterations in the bioavailability of some antimalarials. In-vitro Caco-2 cell monolayers were used here as a model for potential drug absorption related problems and P-gp mediated transport of drugs. Artemisone had the highest permeability at around 50 x 10⁻⁶ cm/sec, followed by amodiaquine around 20 x 10⁻⁶ cm/sec; both mefloquine and artesunate were around 10 x 10⁻⁶ cm/sec. Methylene blue was between 2 and 6 x 10⁻⁶ cm/sec depending on the direction of transport. This 3 fold difference was able to be halved by use of P-gp inhibition. MRP inhibition also assisted the consolidation of the methylene blue transport. Mefloquine was shown to be a P-gp inhibitor affecting our P-gp substrate, Rhodamine 123, although none of the other drugs impacted upon rhodamine123 transport rates. In conclusion, mefloquine is a P-gp inhibitor and methylene blue is a partial substrate; methylene blue may have increased absorption if co-administered with such P-gp inhibitors. An upregulation of P-gp was observed when artemisone and dihydroartemisinin were co-incubated with mefloquine and amodiaquine.