Evolutionary conservation and in vitro reconstitution of microsporidian iron–sulfur cluster biosynthesis

This work was supported by Marie Curie Postdoctoral Fellowships to T.A.W., E. H. and S. L., a European Research Council Advanced Investigator Grant (ERC-2010-AdG-268701) to T.M.E., and a Wellcome Trust Programme Grant (number 045404) to T.M.E. and J.M.L. R.L. acknowledges generous financial support from Deutsche Forschungsgemeinschaft (SFB 593, SFB 987, GRK 1216, LI 415/5), LOEWE program of state Hessen, Max-Planck Gesellschaft, von Behring-Röntgen StiftungMicrosporidians are a diverse group of obligate intracellular parasites that have minimized their genome content and simplified their sub-cellular structures by reductive evolution. Functional studies are limited because we lack reliable genetic tools for their manipulation. Here, we demonstrate that the cristae-deficient mitochondrion (mitosome) of the microsporidian Trachipleistophora hominis is the functional site of iron-sulphur cluster (ISC) assembly, which we suggest is the essential task of this organelle. Cell fractionation, fluorescence imaging and fine-scale immunoelectron microscopy demonstrate that mitosomes contain a complete pathway for [2Fe-2S] cluster biosynthesis that we biochemically reconstituted using purified recombinant mitosomal ISC proteins. Reconstitution proceeded as rapidly and efficiently as observed for yeast or fungal mitochondrial ISC components. Core components of the T. hominis cytosolic iron-sulphur protein assembly (CIA) pathway were also identified including the essential Cfd1-Nbp35 scaffold complex that assembles a [4Fe-4S] cluster as shown by spectroscopic methods in vitro. Phylogenetic analyses reveal that both the ISC and CIA biosynthetic pathways are predominantly bacterial, but their cytosolic and nuclear target Fe/S proteins are mainly archaeal. This mixed evolutionary history of the Fe/S-related proteins and pathways, and their strong conservation among highly reduced parasites, provides additional compelling evidence for the ancient chimeric ancestry of eukaryotes.Publisher PDFPeer reviewe

Phosphate desorption from flooded and reoxidized soils as compared with adsorption characteristics

The purpose of this study was to show how phosphorus (P) desorption can bring further information to what is inferred from adsorption experiments. A simple calculation procedure is proposed to predict the P amount that would be desorbed by washing with salt solution if adsorption was completely reversible. Consequently, the interpretation of P desorption can focus on the irreversible part of adsorption. This approach is applied to samples of acid sulfate soil from Vietnam that have been submerged for different periods of time, at 20 degrees C and 30 degrees C, and also to samples that were reoxidized after flooding. Phosphate desorption linearly increases with P concentration in solution and exponentially decreases with increasing adsorption capacity as expressed by the Freundlich coefficient of adsorption isotherms. These two types of relationships are correctly predicted by our calculation procedure. As far as reversibility is concerned, we find that with respect to calculated desorption, the proportion of irreversible adsorption greatly differ according to treatments. In relative terms, adsorption reversibility is lowest in the reoxidized soils and highest in the wet soils incubated at 30 degrees C. This is related to the type and crystallinity of Fe-oxihydroxides and consequent differences in P-bonding energies

Effect of temperature and flooding duration on phosphate sorption in an acid sulphate soil from Vietnam

Phosphate sorption in soil is controlled largely by Fe-oxihydroxides, and so important changes in P dynamics are expected when the redox potential is modified. Such changes in P sorption when acid soil is flooded, as for rice cultivation, have been evaluated. Samples from an acid sulphate soil in the Mekong Delta of Vietnam were hooded for up to 56 d at 20 degrees C and 30 degrees C. Some of the samples incubated at 30 degrees C were dried in open air for 30 d after Flooding. Small redox potential (Eh 6 were rapidly reached in soil flooded at 30 degrees C; less drastic reducing conditions (Eh congruent to 0.2 V) and pH 4-5 occurred at 20 degrees C. Phosphate sorption increased during flooding. The increase was twofold at 20 degrees C, and 10-fold at 30 degrees C. Phosphate sorption index decreased in the soil that was air dried after flooding at 30 degrees C, but still remained two to three times greater than before flooding. These results were compared to the changes in oxalate-er;tractable Fe, i.e. poorly crystalline or amorphous Fe-oxihydroxides. The increase of P sorption per unit increase of oxalate-Fe was seven to eight fold larger at 30 degrees C than at 20 degrees C

Effect of temperature on rice growth in nutrient solution and in acid sulphate soils from Vietnam

Climatic and soil factors are limiting rice growth in many countries. In Vietnam, a steep gradient of temperature is observed from the North to the South, and acid sulphate soils are frequently devoted to rice production. We have therefore attempted to understand how temperature affects rice growth in these problem soils, by comparison with rice grown in nutrient solution. Two varieties of rice, IR64 and X2, were cultivated in phytotrons at 19/21 degrees C and 28/32 degrees C (day/night) for 56 days, after 3 weeks preculture in optimal conditions. Two soils from the Mekong Delta were tested. Parallel with the growing experiments, these two soils were incubated in order to monitor redox potential (Eh), PH, soluble Al and Fe, soluble, and available P. Tillering retardation at 20 degrees C compared to 30 degrees C was similar in nutrient solutions and in soils. The effect of temperature on increasing plant biomass was more marked in solutions than in soils. The P concentrations in roots and shoots were higher at 20 degrees C than at 30 degrees C, to such an extent that detrimental effect was suspected in plants grown in solution at the lowest temperature. The translocation of Fe from roots to shoots was stimulated upon rising temperature, both in solutions and in soils. This led to plant death on the most acid soil at 30 degrees C. Indeed, the accumulation of Fe in plants grown on soils was enhanced by the release of Fe2+ due to reduction of Fe(III)-oxihydroxides. Severe reducing conditions were created at 30 degrees C: redox potential (Eh) dropped rapidly down to about 0 V. At 20 degrees C, E(h) did not drop below about 0.2 V, which is a value well in the range of Fe(III)/Fe(II) buffering. Parallel to Eh drop, PH increased up to about 6-6.5 at 30 degrees C, which prevented plants from Al toxicity, even in the most acid soil. Phosphate behavior was obviously related to Fe-dynamics: more reducing conditions at 30 degrees C have resulted in enhancement of available P, especially in the most acid soil

Phosphorus sorption in soils of the Mekong delta (Vietnam) as described by the binary Langmuir equation

Phosphate adsorption isotherms were determined for 20, mostly very acidic, soils from the Mekong Delta. The experimental data were well described by a binary Langmuir equation which considers two groups of sorption sites that differ in their P bonding energies. The maximum P-sorption capacities of these sites were related to the soil properties by simple linear correlation and by stepwise multiple regression. Results suggest that high energy sites are on Al-oxihydroxides or small Al-substituted Fe-oxides and, to a lesser extent, on poorly ordered Fe-oxihydroxides. On the other band, the P-sorption capacity of low energy sites is mainly related to clay content, and it increases as pH decreases. These sites are also positively correlated with organic carbon and poorly crystalline Fe-oxihydroxides. However, as these two variables are closely correlated with each other, organic matter is likely to be considered as an indirect factor of P fixation through its association with Fe-oxihydroxides and not as an important source of P-sorption sites. The maximum sorption capacity, i.e. the sum of sorption capacities of the two groups of sites, is well described (r(2) = 0.88) by an equation that takes into account the four variables identified above: Al-bearing oxihydroxides, poorly ordered Fe-oxihydroxides, clay content and pH. Grouping the soils according to the orders of Soil Taxonomy, the P-sorption capacity increases in the following sequence: ultisols < entisols < inceptisols. A P concentration often considered adequate for plant nutrition is 0.2 mg P l(-1) solution, and only the high energy sites are involved in sorption at that concentration. Thus an equation including only Al- and Fe-oxihydroxides could be used to fix P norms in these soils

Ventilator-associated respiratory infection in a resource-restricted setting: impact and etiology

Ventilator-associated respiratory infection (VARI) is a significant problem in resource-restricted intensive care units (ICUs), but differences in casemix and etiology means VARI in resource-restricted ICUs may be different from that found in resource-rich units. Data from these settings are vital to plan preventative interventions and assess their cost-effectiveness, but few are available.We conducted a prospective observational study in four Vietnamese ICUs to assess the incidence and impact of VARI. Patients ≥ 16 years old and expected to be mechanically ventilated > 48 h were enrolled in the study and followed daily for 28 days following ICU admission.Four hundred fifty eligible patients were enrolled over 24 months, and after exclusions, 374 patients' data were analyzed. A total of 92/374 cases of VARI (21.7/1000 ventilator days) were diagnosed; 37 (9.9%) of these met ventilator-associated pneumonia (VAP) criteria (8.7/1000 ventilator days). Patients with any VARI, VAP, or VARI without VAP experienced increased hospital and ICU stay, ICU cost, and antibiotic use (p < 0.01 for all). This was also true for all VARI (p < 0.01 for all) with/without tetanus. There was no increased risk of in-hospital death in patients with VARI compared to those without (VAP HR 1.58, 95% CI 0.75-3.33, p = 0.23; VARI without VAP HR 0.40, 95% CI 0.14-1.17, p = 0.09). In patients with positive endotracheal aspirate cultures, most VARI was caused by Gram-negative organisms; the most frequent were Acinetobacter baumannii (32/73, 43.8%) Klebsiella pneumoniae (26/73, 35.6%), and Pseudomonas aeruginosa (24/73, 32.9%). 40/68 (58.8%) patients with positive cultures for these had carbapenem-resistant isolates. Patients with carbapenem-resistant VARI had significantly greater ICU costs than patients with carbapenem-susceptible isolates (6053 USD (IQR 3806-7824) vs 3131 USD (IQR 2108-7551), p = 0.04) and after correction for adequacy of initial antibiotics and APACHE II score, showed a trend towards increased risk of in-hospital death (HR 2.82, 95% CI 0.75-6.75, p = 0.15).VARI in a resource-restricted setting has limited impact on mortality, but shows significant association with increased patient costs, length of stay, and antibiotic use, particularly when caused by carbapenem-resistant bacteria. Evidence-based interventions to reduce VARI in these settings are urgently needed