Gene cloning, molecular modeling, and phylogenetics of serine protease P32 and serine carboxypeptidase SCP1 from nematophagous fungi Pochonia rubescens and Pochonia chlamydosporia

The fungi Pochonia chlamydosporia and Pochonia rubescens are parasites of nematode eggs and thus are biocontrol agents of nematodes. Proteolytic enzymes such as the S8 proteases VCP1 and P32, secreted during the pathogenesis of nematode eggs, are major virulence factors in these fungi. Recently, expression of these enzymes and of SCP1, a new putative S10 carboxypeptidase, was detected during endophytic colonization of barley roots by these fungi. In our study, we cloned the genomic and mRNA sequences encoding P32 from P. rubescens and SCP1 from P. chlamydosporia. P32 showed a high homology with the serine proteases Pr1A from the entomopathogenic fungus Metarhizium anisopliae and VCP1 from P. chlamydosporia (86% and 76% identity, respectively). However, the catalytic pocket of P32 showed differences in the amino acids of the substrate-recognition sites compared with the catalytic pockets of Pr1A and VCP1 proteases. Phylogenetic analysis of P32 suggests a common ancestor with protease Pr1A. SCP1 displays the characteristic features of a member of the S10 family of serine proteases. Phylogenetic comparisons show that SCP1 and other carboxypeptidases from filamentous fungi have an origin different from that of yeast vacuolar serine carboxypeptidases. Understanding protease genes from nematophagous fungi is crucial for enhancing the biocontrol potential of these organisms.

The imprint of the crustal magnetic field on the thermal spectra and pulse profiles of isolated neutron stars

Isolated neutron stars (NSs) show a bewildering variety of astrophysical manifestations, presumably shaped by the magnetic field strength and topology at birth. Here, using state-of-the-art calculations of the coupled magnetic and thermal evolution of NSs, we compute the thermal spectra and pulse profiles expected for a variety of initial magnetic field configurations. In particular, we contrast models with purely poloidal magnetic fields to models dominated by a strong internal toroidal component. We find that, while the former displays double-peaked profiles and very low pulsed fractions, in the latter, the anisotropy in the surface temperature produced by the toroidal field often results in a single pulse profile, with pulsed fractions that can exceed the 50–60 per cent level even for perfectly isotropic local emission. We further use our theoretical results to generate simulated ‘observed’ spectra, and show that blackbody (BB) fits result in inferred radii that can be significantly smaller than the actual NS radius, even as low as ∼1–2 km for old NSs with strong internal toroidal fields and a high absorption column density along their line of sight. We compute the size of the inferred BB radius for a few representative magnetic field configurations, NS ages and magnitudes of the column density. Our theoretical results are of direct relevance to the interpretation of X-ray observations of isolated NSs, as well as to the constraints on the equation of state of dense matter through radius measurements.

A metabolomic approach to study the rhizodeposition in the tritrophic interaction: tomato, Pochonia chlamydosporia and Meloidogyne javanica

A combined chemometrics-metabolomics approach [excitation–emission matrix (EEM) fluorescence spectroscopy, nuclear magnetic resonance (NMR) and high performance liquid chromatography–mass spectrometry (HPLC–MS)] was used to analyse the rhizodeposition of the tritrophic system: tomato, the plant-parasitic nematode Meloidogyne javanica and the nematode-egg parasitic fungus Pochonia chlamydosporia. Exudates from M. javanica roots were sampled at root penetration (early) and gall development (late). EMM indicated that late root exudates from M. javanica treatments contained more aromatic amino acid compounds than the rest (control, P. chlamydosporia or P. chlamydosporia and M. javanica). 1H NMR showed that organic acids (acetate, lactate, malate, succinate and formic acid) and one unassigned aromatic compound (peak no. 22) were the most relevant metabolites in root exudates. Robust principal component analysis (PCA) grouped early exudates for nematode (PC1) or fungus presence (PC3). PCA found (PC1, 73.31 %) increased acetate and reduced lactate and an unassigned peak no. 22 characteristic of M. javanica root exudates resulting from nematode invasion and feeding. An increase of peak no. 22 (PC3, 4.82 %) characteristic of P. chlamydosporia exudates could be a plant “primer” defence. In late ones in PC3 (8.73 %) the presence of the nematode grouped the samples. HPLC–MS determined rhizosphere fingerprints of 16 (early) and 25 (late exudates) m/z signals, respectively. Late signals were exclusive from M. javanica exudates confirming EEM and 1H NMR results. A 235 m/z signal reduced in M. javanica root exudates (early and late) could be a repressed plant defense. This metabolomic approach and other rhizosphere -omics studies could help to improve plant growth and reduce nematode damage sustainably.

Some isolates of the nematophagous fungus Pochonia chlamydosporia promote root growth and reduce flowering time of tomato

The fungal parasite of nematode eggs Pochonia chlamydosporia is also a root endophyte known to promote growth of some plants. In this study, we analysed the effect of nine P. chlamydosporia isolates from worldwide origin on tomato growth. Experiments were performed at different scales (Petri dish, growth chamber and greenhouse conditions) and developmental stages (seedlings, plantlets and plants). Seven P. chlamydosporia isolates significantly (P < 0.05) increased the number of secondary roots and six of those increased total weight of tomato seedlings. Six P. chlamydosporia isolates also increased root weight of tomato plantlets. Root colonisation varied between different isolates of this fungus. Again P. chlamydosporia significantly increased root growth of tomato plants under greenhouse conditions and reduced flowering and fruiting times (up to 5 and 12 days, respectively) versus uninoculated tomato plants. P. chlamydosporia increased mature fruit weight in tomato plants. The basis of the mechanisms for growth, flowering and yield promotion in tomato by the fungus are unknown. However, we found that P. chlamydosporia can produce Indole-3-acetic acid and solubilise mineral phosphate. These results suggest that plant hormones or nutrient ability could play an important role. Our results put forward the agronomic importance of P. chlamydosporia as biocontrol agent of plant parasitic nematodes with tomato growth promoting capabilities.

The discovery, monitoring and environment of SGR J1935+2154

We report on the discovery of a new member of the magnetar class, SGR J1935+2154, and on its timing and spectral properties measured by an extensive observational campaign carried out between 2014 July and 2015 March with Chandra and XMM–Newton (11 pointings). We discovered the spin period of SGR J1935+2154 through the detection of coherent pulsations at a period of about 3.24 s. The magnetar is slowing down at a rate of P˙=1.43(1)×10−11 s s−1 and with a decreasing trend due to a negative P¨ of −3.5(7) × 10−19 s s−2. This implies a surface dipolar magnetic field strength of ∼2.2 × 1014 G, a characteristic age of about 3.6 kyr and a spin-down luminosity Lsd ∼1.7 × 1034 erg s−1. The source spectrum is well modelled by a blackbody with temperature of about 500 eV plus a power-law component with photon index of about 2. The source showed a moderate long-term variability, with a flux decay of about 25 per cent during the first four months since its discovery, and a re-brightening of the same amount during the second four months. The X-ray data were also used to study the source environment. In particular, we discovered a diffuse emission extending on spatial scales from about 1 arcsec up to at least 1 arcmin around SGR J1935+2154 both in Chandra and XMM–Newton data. This component is constant in flux (at least within uncertainties) and its spectrum is well modelled by a power-law spectrum steeper than that of the pulsar. Though a scattering halo origin seems to be more probable we cannot exclude that part, or all, of the diffuse emission is due to a pulsar wind nebula.

Invisible work, unseen hazards: the health of women immigrant household service workers in Spain

BACKGROUND: Household service work has been largely absent from occupational health studies. We examine the occupational hazards and health effects identified by immigrant women household service workers. METHODS: Exploratory, descriptive study of 46 documented and undocumented immigrant women in household services in Spain, using a phenomenological approach. Data were collected between September 2006 and May 2007 through focus groups and semi-structured individual interviews. Data were separated for analysis by documentation status and sorted using a mixed-generation process. In a second phase of analysis, data on psychosocial hazards were organized using the Copenhagen Psychosocial Questionnaire as a guide. RESULTS: Informants reported a number of environmental, ergonomic and psychosocial hazards and corresponding health effects. Psychosocial hazards were especially strongly present in data. Data on reported hazards were similar by documentation status and varied by several emerging categories: whether participants were primarily cleaners or carers and whether they lived in or outside of the homes of their employers. Documentation status was relevant in terms of empowerment and bargaining, but did not appear to influence work tasks or exposure to hazards directly. CONCLUSIONS: Female immigrant household service workers are exposed to a variety of health hazards that could be acted upon by improved legislation, enforcement, and preventive workplace measures, which are discussed.

Partial rescue of retinal function in chronically hypoglycemic mice

Purpose. Mice rendered hypoglycemic by a null mutation in the glucagon receptor gene Gcgr display late-onset retinal degeneration and loss of retinal sensitivity. Acute hyperglycemia induced by dextrose ingestion does not restore their retinal function, which is consistent with irreversible loss of vision. The goal of this study was to establish whether long-term administration of high dietary glucose rescues retinal function and circuit connectivity in aged Gcgr−/− mice. Methods. Gcgr−/− mice were administered a carbohydrate-rich diet starting at 12 months of age. After 1 month of treatment, retinal function and structure were evaluated using electroretinographic (ERG) recordings and immunohistochemistry. Results. Treatment with a carbohydrate-rich diet raised blood glucose levels and improved retinal function in Gcgr−/− mice. Blood glucose increased from moderate hypoglycemia to euglycemic levels, whereas ERG b-wave sensitivity improved approximately 10-fold. Because the b-wave reflects the electrical activity of second-order cells, we examined for changes in rod-to-bipolar cell synapses. Gcgr−/− retinas have 20% fewer synaptic pairings than Gcgr+/− retinas. Remarkably, most of the lost synapses were located farthest from the bipolar cell body, near the distal boundary of the outer plexiform layer (OPL), suggesting that apical synapses are most vulnerable to chronic hypoglycemia. Although treatment with the carbohydrate-rich diet restored retinal function, it did not restore these synaptic contacts. Conclusions. Prolonged exposure to diet-induced euglycemia improves retinal function but does not reestablish synaptic contacts lost by chronic hypoglycemia. These results suggest that retinal neurons have a homeostatic mechanism that integrates energetic status over prolonged periods of time and allows them to recover functionality despite synaptic loss.

Unifying the observational diversity of isolated neutron stars via magneto-thermal evolution models

Observations of magnetars and some of the high magnetic field pulsars have shown that their thermal luminosity is systematically higher than that of classical radio-pulsars, thus confirming the idea that magnetic fields are involved in their X-ray emission. Here we present the results of 2D simulations of the fully coupled evolution of temperature and magnetic field in neutron stars, including the state-of-the-art kinetic coefficients and, for the first time, the important effect of the Hall term. After gathering and thoroughly re-analysing in a consistent way all the best available data on isolated, thermally emitting neutron stars, we compare our theoretical models to a data sample of 40 sources. We find that our evolutionary models can explain the phenomenological diversity of magnetars, high-B radio-pulsars, and isolated nearby neutron stars by only varying their initial magnetic field, mass and envelope composition. Nearly all sources appear to follow the expectations of the standard theoretical models. Finally, we discuss the expected outburst rates and the evolutionary links between different classes. Our results constitute a major step towards the grand unification of the isolated neutron star zoo.

The outburst decay of the low magnetic field magnetar SGR 0418+5729

We report on the long-term X-ray monitoring of the outburst decay of the low magnetic field magnetar SGR 0418+5729 using all the available X-ray data obtained with RXTE, Swift, Chandra, and XMM-Newton observations from the discovery of the source in 2009 June up to 2012 August. The timing analysis allowed us to obtain the first measurement of the period derivative of SGR 0418+5729: ˙ P = 4(1) × 10−15 s s−1, significant at a ∼3.5σ confidence level. This leads to a surface dipolar magnetic field of Bdip 6 × 1012 G. This measurement confirms SGR 0418+5729 as the lowest magnetic field magnetar. Following the flux and spectral evolution from the beginning of the outburst up to ∼1200 days, we observe a gradual cooling of the tiny hot spot responsible for the X-ray emission, from a temperature of ∼0.9 to 0.3 keV. Simultaneously, the X-ray flux decreased by about three orders of magnitude: from about 1.4 × 10−11 to 1.2 × 10−14 erg s−1 cm−2. Deep radio, millimeter, optical, and gamma-ray observations did not detect the source counterpart, implying stringent limits on its multi-band emission, as well as constraints on the presence of a fossil disk. By modeling the magneto-thermal secular evolution of SGR 0418+5729, we infer a realistic age of ∼550 kyr, and a dipolar magnetic field at birth of ∼1014 G. The outburst characteristics suggest the presence of a thin twisted bundle with a small heated spot at its base. The bundle untwisted in the first few months following the outburst, while the hot spot decreases in temperature and size. We estimate the outburst rate of low magnetic field magnetars to be about one per year per galaxy, and we briefly discuss the consequences of such a result in several other astrophysical contexts.

A new low magnetic field magnetar: the 2011 outburst of Swift J1822.3–1606

We report on the long-term X-ray monitoring with Swift, RXTE, Suzaku, Chandra, and XMM-Newton of the outburst of the newly discovered magnetar Swift J1822.3–1606 (SGR 1822–1606), from the first observations soon after the detection of the short X-ray bursts which led to its discovery, through the first stages of its outburst decay (covering the time span from 2011 July until the end of 2012 April). We also report on archival ROSAT observations which detected the source during its likely quiescent state, and on upper limits on Swift J1822.3–1606's radio-pulsed and optical emission during outburst, with the Green Bank Telescope and the Gran Telescopio Canarias, respectively. Our X-ray timing analysis finds the source rotating with a period of P = 8.43772016(2) s and a period derivative P = 8.3(2)×10−14 s s−1, which implies an inferred dipolar surface magnetic field of B sime 2.7 × 1013 G at the equator. This measurement makes Swift J1822.3–1606 the second lowest magnetic field magnetar (after SGR 0418+5729). Following the flux and spectral evolution from the beginning of the outburst, we find that the flux decreased by about an order of magnitude, with a subtle softening of the spectrum, both typical of the outburst decay of magnetars. By modeling the secular thermal evolution of Swift J1822.3–1606, we find that the observed timing properties of the source, as well as its quiescent X-ray luminosity, can be reproduced if it was born with a poloidal and crustal toroidal fields of Bp ~ 1.5 × 1014 G and B tor ~ 7 × 1014 G, respectively, and if its current age is ~550 kyr.

Spectral features in isolated neutron stars induced by inhomogeneous surface temperatures

The thermal X-ray spectra of several isolated neutron stars display deviations from a pure blackbody. The accurate physical interpretation of these spectral features bears profound implications for our understanding of the atmospheric composition, magnetic field strength and topology, and equation of state of dense matter. With specific details varying from source to source, common explanations for the features have ranged from atomic transitions in the magnetized atmospheres or condensed surface, to cyclotron lines generated in a hot ionized layer near the surface. Here, we quantitatively evaluate the X-ray spectral distortions induced by inhomogeneous temperature distributions of the neutron star surface. To this aim, we explore several surface temperature distributions, we simulate their corresponding general relativistic X-ray spectra (assuming an isotropic, blackbody emission), and fit the latter with a single blackbody model. We find that, in some cases, the presence of a spurious ‘spectral line’ is required at a high significance level in order to obtain statistically acceptable fits, with central energy and equivalent width similar to the values typically observed. We also perform a fit to a specific object, RX J0806.4−4123, finding several surface temperature distributions able to model the observed spectrum. The explored effect is unlikely to work in all sources with detected lines, but in some cases it can indeed be responsible for the appearance of such lines. Our results enforce the idea that surface temperature anisotropy can be an important factor that should be considered and explored also in combination with more sophisticated emission models like atmospheres.

PipX, the coactivator of NtcA, is a global regulator in cyanobacteria

To modulate the expression of genes involved in nitrogen assimilation, the cyanobacterial PII-interacting protein X (PipX) interacts with the global transcriptional regulator NtcA and the signal transduction protein PII, a protein found in all three domains of life as an integrator of signals of the nitrogen and carbon balance. PipX can form alternate complexes with NtcA and PII, and these interactions are stimulated and inhibited, respectively, by 2-oxoglutarate, providing a mechanistic link between PII signaling and NtcA-regulated gene expression. Here, we demonstrate that PipX is involved in a much wider interaction network. The effect of pipX alleles on transcript levels was studied by RNA sequencing of S. elongatus strains grown in the presence of either nitrate or ammonium, followed by multivariate analyses of relevant mutant/control comparisons. As a result of this process, 222 genes were classified into six coherent groups of differentially regulated genes, two of which, containing either NtcA-activated or NtcA-repressed genes, provided further insights into the function of NtcA–PipX complexes. The remaining four groups suggest the involvement of PipX in at least three NtcA-independent regulatory pathways. Our results pave the way to uncover new regulatory interactions and mechanisms in the control of gene expression in cyanobacteria.

Semantic change and intersubjectification: The origin of reprise evidential conditional in Old Catalan

The reprise evidential conditional (REC) is nowadays not very usual in Catalan: it is restricted to journalistic language and to some very formal genres (such as academic or legal language), it is not present in spontaneous discourse. On the one hand, it has been described among the rather new modality values of the conditional. On the other, the normative tradition tended to reject it for being a gallicism, or to describe it as an unsuitable neologism. Thanks to the extraction from text corpora, we surprisingly find this REC in Catalan from the beginning of the fourteenth century to the contemporary age, with semantic and pragmatic nuances and different evidence of grammaticalization. Due to the current interest in evidentiality, the REC has been widely studied in French, Italian and Portuguese, focusing mainly on its contemporary uses and not so intensively on the diachronic process that could explain the origin of this value. In line with this research, that we initiated studying the epistemic and evidential future in Catalan, our aim is to describe: a) the pragmatic context that could have been the initial point of the REC in the thirteenth century, before we find indisputable attestations of this use; b) the path of semantic change followed by the conditional from a ‘future in the past’ tense to the acquisition of epistemic and evidential values; and c) the role played by invited inferences, subjectification and intersubjectification in this change.