Human CLPP reverts the longevity phenotype of a fungal ClpP deletion strain

Mitochondrial maintenance crucially depends on the quality control of proteins by various chaperones, proteases and repair enzymes. While most of the involved components have been studied in some detail, little is known on the biological role of the CLPXP protease complex located in the mitochondrial matrix. Here we show that deletion of PaClpP, encoding the CLP protease proteolytic subunit CLPP, leads to an unexpected healthy phenotype and increased lifespan of the fungal ageing model organism Podospora anserina. This phenotype can be reverted by expression of human ClpP in the fungal deletion background, demonstrating functional conservation of human and fungal CLPP. Our results show that the biological role of eukaryotic CLP proteases can be studied in an experimentally accessible model organism

A differential genome-wide transcriptome analysis : impact of cellular copper on complex biological processes like aging and development

The regulation of cellular copper homeostasis is crucial in biology. Impairments lead to severe dysfunctions and are known to affect aging and development. Previously, a loss-of-function mutation in the gene encoding the copper-sensing and copper-regulated transcription factor GRISEA of the filamentous fungus Podospora anserina was reported to lead to cellular copper depletion and a pleiotropic phenotype with hypopigmentation of the mycelium and the ascospores, affected fertility and increased lifespan by approximately 60% when compared to the wild type. This phenotype is linked to a switch from a copper-dependent standard to an alternative respiration leading to both a reduced generation of reactive oxygen species (ROS) and of adenosine triphosphate (ATP). We performed a genome-wide comparative transcriptome analysis of a wild-type strain and the copper-depleted grisea mutant. We unambiguously assigned 9,700 sequences of the transcriptome in both strains to the more than 10,600 predicted and annotated open reading frames of the P. anserina genome indicating 90% coverage of the transcriptome. 4,752 of the transcripts differed significantly in abundance with 1,156 transcripts differing at least 3-fold. Selected genes were investigated by qRT-PCR analyses. Apart from this general characterization we analyzed the data with special emphasis on molecular pathways related to the grisea mutation taking advantage of the available complete genomic sequence of P. anserina. This analysis verified but also corrected conclusions from earlier data obtained by single gene analysis, identified new candidates of factors as part of the cellular copper homeostasis system including target genes of transcription factor GRISEA, and provides a rich reference source of quantitative data for further in detail investigations. Overall, the present study demonstrates the importance of systems biology approaches also in cases were mutations in single genes are analyzed to explain the underlying mechanisms controlling complex biological processes like aging and development

Practices of Ethnographic Research: Introduction to the Special Issue

[no abstract available

IC10~X-1/NGC300~X-1: the very immediate progenitors of BH-BH binaries

We investigate the future evolution of two extragalactic X-ray binaries: IC10 X-1 and NGC300 X-1. Each of them consists of a high mass BH (\sim 20-30 \msun) accreting from a massive WR star companion (\gtrsim 20 \msun), and both are located in low metallicity galaxies. We analyze the current state of the systems and demonstrate that both systems will very quickly ($\lesssim 0.3$ Myr) form close BH-BH binaries with the short coalescence time ($\sim 3$ Gyr) and large chirp mass (\sim 15 \msun). The formation of BH-BH system seems unavoidable, as {\em (i)} WR companions are well within their Roche lobes and they do not expand so no Roche lobe overflow is expected, {\em (ii)} even intense WR wind mass loss does not remove sufficient mass to prohibit the formation of the second BH, {\em (ii)} even if BH receives the large natal kick, the systems are very closely bound and are almost impossible to disrupt. As there are two such immediate BH-BH progenitor systems within 2 Mpc and as the current gravitational wave instruments LIGO/VIRGO (initial stage) can detect such massive BH-BH mergers out to $\sim 200$ Mpc, the empirically estimated detection rate of such inspirals is $R=3.36^{+8.29}_{-2.92}$ at the 99% confidence level. If there is no detection in the current LIGO/VIRGO data (unreleased year of $s6$ run), the existence of these two massive BH systems poses an interesting challenge. Either the gravitational radiation search is not sensitive to massive inspirals or there is some fundamental misunderstanding of stellar evolution physics leading directly to the formation of BH-BH binaries.Comment: 9 pages, resubmitted to ApJ with major extensio

Macroclumping as solution of the discrepancy between Hα and P v mass loss diagnostics for O-type stars

Context. Recent studies of O-type stars have demonstrated that discrepant mass-loss rates are obtained when different diagnostic methods are employed. Fitting the unsaturated UV resonance lines (e.g., P v) gives drastically lower values than obtained from the Hα emission. Wind inhomogeneity (so-called "clumping") may be the main cause of this discrepancy. Aims. In a previous paper, we presented 3D Monte-Carlo calculations for the formation of scattering lines in a clumped stellar wind. In the present paper we select five O-type supergiants (from O4 to O7) and test whether the reported discrepancies can be resolved this way. Methods. In the first step, the analyses started with simulating the observed spectra with Potsdam Wolf-Rayet (PoWR) non-LTE model atmospheres. The mass-loss rates are adjusted to fit to the observed Hα emission lines best. For the unsaturated UV resonance lines (i.e., P v) we then applied our 3D Monte-Carlo code, which can account for wind clumps of any optical depths ("macroclumping"), a non-void interclump medium, and a velocity dispersion inside the clumps. The ionization stratifications and underlying photospheric spectra were adopted from the PoWR models. The properties of the wind clumps were constrained by fitting the observed resonance line profiles. Results. Our results show that with the mass-loss rates that fit Hα (and other Balmer and He ii lines), the UV resonance lines (especially the unsaturated doublet of P v) can also be reproduced with no problem when macroclumping is taken into account. There is no need to artificially reduce the mass-loss rates or to assume a subsolar phosphorus abundance or an extremely high clumping factor, unlike what was claimed by other authors. These consistent mass-loss rates are lower by a factor of 1.3 to 2.6, compared to the mass-loss rate recipe from Vink et al. Conclusions. Macroclumping resolves the previously reported discrepancy between Hα and P v mass-loss diagnostics.Facultad de Ciencias Astronómicas y GeofísicasInstituto de Astrofísica de La Plat

Macroclumping as solution of the discrepancy between Hα and P v mass loss diagnostics for O-type stars

Context. Recent studies of O-type stars have demonstrated that discrepant mass-loss rates are obtained when different diagnostic methods are employed. Fitting the unsaturated UV resonance lines (e.g., P v) gives drastically lower values than obtained from the Hα emission. Wind inhomogeneity (so-called "clumping") may be the main cause of this discrepancy. Aims. In a previous paper, we presented 3D Monte-Carlo calculations for the formation of scattering lines in a clumped stellar wind. In the present paper we select five O-type supergiants (from O4 to O7) and test whether the reported discrepancies can be resolved this way. Methods. In the first step, the analyses started with simulating the observed spectra with Potsdam Wolf-Rayet (PoWR) non-LTE model atmospheres. The mass-loss rates are adjusted to fit to the observed Hα emission lines best. For the unsaturated UV resonance lines (i.e., P v) we then applied our 3D Monte-Carlo code, which can account for wind clumps of any optical depths ("macroclumping"), a non-void interclump medium, and a velocity dispersion inside the clumps. The ionization stratifications and underlying photospheric spectra were adopted from the PoWR models. The properties of the wind clumps were constrained by fitting the observed resonance line profiles. Results. Our results show that with the mass-loss rates that fit Hα (and other Balmer and He ii lines), the UV resonance lines (especially the unsaturated doublet of P v) can also be reproduced with no problem when macroclumping is taken into account. There is no need to artificially reduce the mass-loss rates or to assume a subsolar phosphorus abundance or an extremely high clumping factor, unlike what was claimed by other authors. These consistent mass-loss rates are lower by a factor of 1.3 to 2.6, compared to the mass-loss rate recipe from Vink et al. Conclusions. Macroclumping resolves the previously reported discrepancy between Hα and P v mass-loss diagnostics.Facultad de Ciencias Astronómicas y GeofísicasInstituto de Astrofísica de La Plat

Deep transformation models for functional outcome prediction after acute ischemic stroke

In many medical applications, interpretable models with high prediction performance are sought. Often, those models are required to handle semi-structured data like tabular and image data. We show how to apply deep transformation models (DTMs) for distributional regression which fulfill these requirements. DTMs allow the data analyst to specify (deep) neural networks for different input modalities making them applicable to various research questions. Like statistical models, DTMs can provide interpretable effect estimates while achieving the state-of-the-art prediction performance of deep neural networks. In addition, the construction of ensembles of DTMs that retain model structure and interpretability allows quantifying epistemic and aleatoric uncertainty. In this study, we compare several DTMs, including baseline-adjusted models, trained on a semi-structured data set of 407 stroke patients with the aim to predict ordinal functional outcome three months after stroke. We follow statistical principles of model-building to achieve an adequate trade-off between interpretability and flexibility while assessing the relative importance of the involved data modalities. We evaluate the models for an ordinal and dichotomized version of the outcome as used in clinical practice. We show that both, tabular clinical and brain imaging data, are useful for functional outcome prediction, while models based on tabular data only outperform those based on imaging data only. There is no substantial evidence for improved prediction when combining both data modalities. Overall, we highlight that DTMs provide a powerful, interpretable approach to analyzing semi-structured data and that they have the potential to support clinical decision making.Comment: Preprint under revie

Macroclumping as solution of the discrepancy between Hα and P v mass loss diagnostics for O-type stars

Context. Recent studies of O-type stars have demonstrated that discrepant mass-loss rates are obtained when different diagnostic methods are employed. Fitting the unsaturated UV resonance lines (e.g., P v) gives drastically lower values than obtained from the Hα emission. Wind inhomogeneity (so-called "clumping") may be the main cause of this discrepancy. Aims. In a previous paper, we presented 3D Monte-Carlo calculations for the formation of scattering lines in a clumped stellar wind. In the present paper we select five O-type supergiants (from O4 to O7) and test whether the reported discrepancies can be resolved this way. Methods. In the first step, the analyses started with simulating the observed spectra with Potsdam Wolf-Rayet (PoWR) non-LTE model atmospheres. The mass-loss rates are adjusted to fit to the observed Hα emission lines best. For the unsaturated UV resonance lines (i.e., P v) we then applied our 3D Monte-Carlo code, which can account for wind clumps of any optical depths ("macroclumping"), a non-void interclump medium, and a velocity dispersion inside the clumps. The ionization stratifications and underlying photospheric spectra were adopted from the PoWR models. The properties of the wind clumps were constrained by fitting the observed resonance line profiles. Results. Our results show that with the mass-loss rates that fit Hα (and other Balmer and He ii lines), the UV resonance lines (especially the unsaturated doublet of P v) can also be reproduced with no problem when macroclumping is taken into account. There is no need to artificially reduce the mass-loss rates or to assume a subsolar phosphorus abundance or an extremely high clumping factor, unlike what was claimed by other authors. These consistent mass-loss rates are lower by a factor of 1.3 to 2.6, compared to the mass-loss rate recipe from Vink et al. Conclusions. Macroclumping resolves the previously reported discrepancy between Hα and P v mass-loss diagnostics.Facultad de Ciencias Astronómicas y GeofísicasInstituto de Astrofísica de La Plat

Integrin αE(CD103) Is Involved in Regulatory T-Cell Function in Allergic Contact Hypersensitivity

Murine contact hypersensitivity (CHS) is a dendritic cell (DC)-dependent T-cell-mediated inflammation with CD8+ T cells as effectors and CD4+ T cells as regulators (Treg cells) that models human allergic contact dermatitis. The integrin αE(CD103) is expressed by some T-cell and DC subsets and has been implicated in epithelial lymphocyte localization, but its role in immune regulation remains enigmatic. We have identified a function for CD103 in the development of cutaneous allergic immune responses. CHS responses, but not irritant contact dermatitis, were significantly augmented in CD103-deficient mice in hapten-challenged skin. Phenotype and function of skin DCs during sensitization were normal, whereas adoptive transfer experiments revealed that the elevated CHS response in CD103-deficient mice is transferred by primed T cells and is independent of resident cells in recipient mice. While T-cell counts were elevated in challenged skin of CD103-deficient mice, the FoxP3 expression level of CD4+CD25+ Treg cells was significantly reduced, indicating impaired functionality. Indeed, Treg cells from CD103-deficient mice were not able to suppress CHS reactions during the elicitation phase. Further, CD103 on FoxP3+ Treg cells was involved in Treg retention to inflamed skin. These findings indicate an unexpected dichotomous functional role for CD103 on Treg cells by modulating FoxP3 expression

Crystal Structure of the C-type Lectin-like Domain from the Human Hematopoietic Cell Receptor CD69

CD69, one of the earliest specific antigens acquired during lymphoid activation, acts as a signal-transducing receptor involved in cellular activation events, including proliferation and induction of specific genes. CD69 belongs to a family of receptors that modulate the immune response and whose genes are clustered in the natural killer (NK) gene complex. The extracellular portion of these receptors represent a subfamily of C-type lectin-like domains (CTLDs), which are divergent from true C-type lectins and are referred to as NK-cell domains (NKDs). We have determined the three-dimensional structure of human CD69 NKD in two different crystal forms. CD69 NKD adopts the canonical CTLD fold but lacks the features involved in Ca(2+) and carbohydrate binding by C-type lectins. CD69 NKD dimerizes noncovalently, both in solution and in crystalline state. The dimer interface consists of a hydrophobic, loosely packed core, surrounded by polar interactions, including an interdomain beta sheet. The intersubunit core shows certain structural plasticity that may facilitate conformational rearrangements for binding to ligands. The surface equivalent to the binding site of other members of the CTLD superfamily reveals a hydrophobic patch surrounded by conserved charged residues that probably constitutes the CD69 ligand-binding site.Fil: Llera, Andrea Sabina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Viedma, Fernando. Universidad Autónoma de Madrid; EspañaFil: Sánchez Madrid, Francisco. Universidad Autónoma de Madrid; EspañaFil: Tormo, José. Universidad Autónoma de Madrid; Españ