Unsaturated glycerophospholipids mediate heme crystallization: biological implications for hemozoin formation in the kissing bug Rhodnius prolixus

Hemozoin (Hz) is a heme crystal produced by some blood-feeding organisms, as an efficient way to detoxify heme derived from hemoglobin digestion. In the triatomine insect Rhodnius prolixus , Hz is essentially produced by midgut extracellular phospholipid membranes known as perimicrovillar membranes (PMVM). Here, we investigated the role of commercial glycerophospholipids containing serine, choline and ethanolamine as headgroups and R. prolixus midgut lipids (RML) in heme crystallization. All commercial unsaturated forms of phospholipids, as well as RML, mediated fast and efficient β-hematin formation by means of two kinetically distinct mechanisms: an early and fast component, followed by a late and slow one. The fastest reactions observed were induced by unsaturated forms of phosphatidylethanolamine (uPE) and phosphatidylcholine (uPC), with half-lives of 0.04 and 0.7 minutes, respectively. β-hematin crystal morphologies were strikingly distinct among groups, with uPE producing homogeneous regular brick-shaped crystals. Interestingly, uPC-mediated reactions resulted in two morphologically distinct crystal populations: one less representative group of regular crystals, resembling those induced by uPE, and the other largely represented by crystals with numerous sharp edges and tapered ends. Heme crystallization reactions induced by RML were efficient, with a heme to β-hematin conversion rate higher than 70%, but clearly slower (t1/2 of 9.9-17.7 minutes) than those induced by uPC and uPE. Interestingly, crystals produced by RML were homogeneous in shape and quite similar to those mediated by uPE. Thus, β-hematin formation can be rapidly and efficiently induced by unsaturated glycerophospholipids, particularly uPE and uPC, and may play a role on biological heme crystallization in R. prolixus midgut

Distribution of dark and baryonic matter in clusters of galaxies

We present the analysis of baryonic and non-baryonic matter distribution in a sample of ten nearby clusters (0.03<z<0.09) with temperatures between 4.7 and 9.4 keV. These galaxy clusters are studied in detail using X-ray data and global physical properties are determined. Correlations between these quantities are analysed and compared with the results for distant clusters. We find an interesting correlation between the extent of the intra-cluster gas relative to the dark matter distribution. The extent of the gas relative to the extent of the dark matter tends to be larger in less massive clusters. This correlation might give us some hints on non-gravitational processes in clusters. We do not see evolution in the gas mass fraction out to a redshift of unity. Within r_{500}, the mean gas mass fraction obtained is 0.16\pm0.02 h_{50}^{-3/2}.Comment: 11 pages, 11 figures, A&A in pres

The dynamical state of the Coma cluster with XMM-Newton

We present in this paper a substructure and spectroimaging study of the Coma cluster of galaxies based on XMM-Newton data. XMM-Newton performed a mosaic of observations of Coma to ensure a large coverage of the cluster. We add the different pointings together and fit elliptical beta-models to the data. We subtract the cluster models from the data and look for residuals, which can be interpreted as substructure. We find several significant structures: the well-known subgroup connected to NGC4839 in the South-West of the cluster, and another substructure located between NGC 4839 and the centre of the Coma cluster. Constructing a hardness ratio image, which can be used as a temperature map we see that in front of this new structure the temperature is significantly increased (higher or equal 10 keV). We interpret this temperature enhancement as the result of heating as this structure falls onto the Coma cluster. We furthermore reconfirm the filament-like structure South-East of the cluster centre. This region is significantly cooler than the mean cluster temperature. We estimate the temperature of this structure to be equal or below 1keV. A possible scenario to explain the observed features is stripping caused by the infall of a small group of galaxies located around the two galaxies NGC4921 and NGC4911 into the Coma cluster with a non-zero impact parameter. We also see significant X-ray depressions North and South-East of NGC4921, which might either be linked to tidal forces due to the merger with the Western structure or connected to an older cluster merger.Comment: 12 pages, 4 figures accepted for publication in A&

An XMM-Newton study of the RGH 80 galaxy group

We present an X-ray study of the galaxy group RGH 80, observed by XMM-Newton. The X-ray emission of the gas is detected out to ~ 462h^{-1}_{50} kpc, corresponding to ~ 0.45 r_{200}. The group is relatively gas rich and luminous with respect to its temperature of 1.01 +/- 0.01 keV. Using the deprojected spectral analysis, we find that the temperature peaks at ~ 1.3 keV around 0.11r_{200}, and then decreases inwards to 0.83 keV at the center and outwards to ~ 70% of the peak value at large radii. Within the central ~ 60 kpc of the group where the gas cooling time is less than the Hubble time, two-temperature model with temperatures of 0.82 and 1.51 keV and the Galactic absorption gives the best fit of the spectra, with ~ 20% volume occupied by the cool component. We also derive the gas entropy distribution, which is consistent with the prediction of cooling and/or internal heating models. Furthermore, the abundances of O, Mg, Si, S, and Fe decrease monotonically with radius. With the observed abundance ratio pattern, we estimate that ~ 85% or ~ 72% of the iron mass is contributed by SN Ia, depending on the adopted SN II models.Comment: 14 pages, 9 figures, accepted for publication in A&

Spatially resolved X-ray spectroscopy of cooling clusters of galaxies

We present spatially resolved X-ray spectra taken with the EPIC cameras of XMM-Newton of a sample of 17 cooling clusters and three non-cooling clusters for comparison. The deprojected spectra are analyzed with a multi-temperature model, independent of any a priori assumptions about the physics behind the cooling and heating mechanisms. All cooling clusters show a central decrement of the average temperature, most of them of a factor of ~2. At each radius within the cooling region the gas is non-isothermal. The differential emission measure distribution peaks near the maximum (ambient) temperature, and steeply declines towards lower temperatures, proportional to T^3, or alternatively a cut-off at about a quarter to half of the maximum temperature. In general, we find a poor correlation between radio flux of the central galaxy and the temperature decrement of the cooling flow. This is interpreted as evidence that except for a few cases heating by a central AGN is not the most common cause of weak cooling flows. We investigate the role of heat conduction by electrons and find that the theoretically predicted conductivity rates are not high enough to balance radiation losses. The differential emission measure distribution has remarkable similarities with the predictions from coronal magnetic loop models. Also the physical processes involved (radiative cooling, thermal conduction along the loops, gravity) are similar for clusters loops and coronal loops. If coronal loop models apply to clusters, we find that a few hundred loops per scale height should be present. The typical loop sizes deduced from the observed emission measure distribution are consistent with the characteristic magnetic field sizes deduced from Faraday rotation measurements.Comment: Accepted for publication in Astronomy and Astrophysics, main journal, 25 pages, 12 figure

A Chandra and XMM View of the Mass & Metals in Galaxy Groups and Clusters

X-ray observations with Chandra and XMM are providing valuable new measurements of the baryonic and dark matter content of groups and clusters. Masses of cD clusters obtained from X-ray and gravitational lensing studies generally show good agreement, therefore providing important validation of both methods. Gas fractions have been obtained for several clusters that verify previous results for a low matter density (Omega_m ~0.3). Chandra has also provided measurements of the mass profiles deep down into several cluster cores and has generally found no significant deviations from CDM predictions in contrast to the flat core density profiles inferred from the rotation curves of low-surface brightness galaxies and dwarf galaxies; i.e., there is no evidence for self-interacting dark matter in cluster cores. Finally, initial studies of the iron and silicon abundances in centrally E-dominated groups show that they have pronounced gradients from 1-2 solar values within the central 30-50 kpc that fall to values of 0.3-0.5 solar at larger radii. The Si/Fe ratios are consistent with approximately 80% of the metals originating from Type Ia supernovae. Several cD clusters also display central Fe enhancements suggestive of Type Ia supernova enrichment, though some have central dips that may provide a vital clue for solving the cooling flow mystery

Silencing of the 20S proteasomal subunit-α6 triggers full oogenesis arrest and increased mRNA levels of the selective autophagy adaptor protein p62/SQSTM1 in the ovary of the vector Rhodnius prolixus.

The high reproductive rates of insects contribute significantly to their ability to act as vectors of a variety of vector-borne diseases. Therefore, it is strategically critical to find molecular targets with biotechnological potential through the functional study of genes essential for insect reproduction. The ubiquitin-proteasome system is a vital degradative pathway that contributes to the maintenance of regular eukaryotic cell proteostasis. This mechanism involves the action of enzymes to covalently link ubiquitin to proteins that are meant to be delivered to the 26S proteasome and broken down. The 26S proteasome is a large protease complex (including the 20S and 19S subcomplexes) that binds, deubiquitylates, unfolds, and degrades its substrates. Here, we used bioinformatics to identify the genes that encode the seven α and β subunits of the 20S proteasome in the genome of R. prolixus and learned that those transcripts are accumulated into mature oocytes. To access proteasome function during oogenesis, we conducted RNAi functional tests employing one of the 20S proteasome subunits (Prosα6) as a tool to suppress 20S proteasomal activity. We found that Prosα6 silencing resulted in no changes in TAG buildup in the fat body and unaffected availability of yolk proteins in the hemolymph of vitellogenic females. Despite this, the silencing of Prosα6 culminated in the impairment of oocyte maturation at the early stages of oogenesis. Overall, we discovered that proteasome activity is especially important for the signals that initiate oogenesis in R. prolixus and discuss in what manner further investigations on the regulation of proteasome assembly and activity might contribute to the unraveling of oogenesis molecular mechanisms and oocyte maturation in this vector

Expression of acyl-CoA-binding protein 5 from Rhodnius prolixus and its inhibition by RNA interference.

The acyl-CoA-binding proteins (ACBP) act by regulating the availability of acyl-CoA in the cytoplasm and must have essential functions in lipid metabolism. The genome of the kissing-bug Rhodnius prolixus encodes five proteins of this family, but little is known about them. In this study we investigated the expression and function of RpACBP-5. Feeding induced RpACBP-5 gene expression in the posterior midgut, and an increase of about four times was observed two days after the blood meal. However, the amount of protein, which was only detected in this organ, did not change during digestion. The RpACBP-5 gene was also highly expressed in pre-vitellogenic and vitellogenic oocytes. Recombinant RpACBP-5 was shown to bind to acyl-CoA of different lengths, and it exhibited nanomolar affinity to lauroyl-CoA in an isothermal titration assay, indicating that RpACBP-5 is a functional ACBP. RpACBP-5 knockdown by RNA interference did not affect digestion, egg laying and hatching, survival, or accumulation of triacylglycerol in the fat body and oocytes. Similarly, double knockdown of RpACBP-1 and RpACBP-5 did not alter egg laying and hatching, survival, accumulation of triacylglycerol in the fat body and oocytes, or the neutral lipid composition of the posterior midgut or hemolymph. These results show that RpACBP-5 is a functional ACBP but indicate that the lack of a detectable phenotype in the knockdown insects may be a consequence of functional overlap of the proteins of the ACBP family found in the insect

Structural and physico-chemical features of the glycerophospholipids investigated in the present work.

a<p>T_m values were obtained from the literature <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088976#pone.0088976-Silvius1" target="_blank">[67]</a>.</p