Biogenesis of the inner membrane complex is dependent on vesicular transport by the alveolate specific GTPase Rab11B

Apicomplexan parasites belong to a recently recognised group of protozoa referred to as Alveolata. These protists contain membranous sacs (alveoli) beneath the plasma membrane, termed the Inner Membrane Complex (IMC) in the case of Apicomplexa. During parasite replication the IMC is formed de novo within the mother cell in a process described as internal budding. We hypothesized that an alveolate specific factor is involved in the specific transport of vesicles from the Golgi to the IMC and identified the small GTPase Rab11B as an alveolate specific Rab-GTPase that localises to the growing end of the IMC during replication of Toxoplasma gondii. Conditional interference with Rab11B function leads to a profound defect in IMC biogenesis, indicating that Rab11B is required for the transport of Golgi derived vesicles to the nascent IMC of the daughter cell. Curiously, a block in IMC biogenesis did not affect formation of sub-pellicular microtubules, indicating that IMC biogenesis and formation of sub-pellicular microtubules is not mechanistically linked. We propose a model where Rab11B specifically transports vesicles derived from the Golgi to the immature IMC of the growing daughter parasites

Probing the spectral shape of dust emission with the DustPedia galaxy sample

The objective of this paper is to understand the variance of the far-infrared (FIR) spectral energy distribution (SED) of the DustPedia galaxies, and its link with the stellar and dust properties. An interesting aspect of the dust emission is the inferred FIR colours which could inform us about the dust content of galaxies, and how it varies with the physical conditions within galaxies. However, the inherent complexity of dust grains as well as the variety of physical properties depending on dust, hinder our ability to utilise their maximum potential. We use principal component analysis (PCA) to explore new hidden correlations with many relevant physical properties such as the dust luminosity, dust temperature, dust mass, bolometric luminosity, star-formation rate (SFR), stellar mass, specific SFR, dust-to-stellar mass ratio, the fraction of absorbed stellar luminosity by dust (f_abs), and metallicity. We find that 95% of the variance in our sample can be described by two principal components (PCs). The first component controls the wavelength of the peak of the SED, while the second characterises the width. The physical quantities that correlate better with the coefficients of the first two PCs, and thus control the shape of the FIR SED are: the dust temperature, the dust luminosity, the SFR, and f_abs. Finally, we find a weak tendency for low-metallicity galaxies to have warmer and broader SEDs, while on the other hand high-metallicity galaxies have FIR SEDs that are colder and narrower

High-resolution, 3D radiative transfer modelling : V. A detailed model of the M 51 interacting pair

Context. Investigating the dust heating mechanisms in galaxies provides a deeper understanding of how the internal energy balance drives their evolution. Over the last decade radiative transfer simulations based on the Monte Carlo method have emphasised the role of the various stellar populations heating the diffuse dust. Beyond the expected heating through ongoing star formation, older stellar populations (>= 8 Gyr) and even active galactic nuclei can both contribute energy to the infrared emission of diffuse dust.Aims. In this particular study we examine how the radiation of an external heating source, such as the less massive galaxy NGC 5195 in the M 51 interacting system, could affect the heating of the diffuse dust of its parent galaxy NGC 5194, and vice versa. Our goal is to quantify the exchange of energy between the two galaxies by mapping the 3D distribution of their radiation field.Methods. We used SKIRT, a state-of-the-art 3D Monte Carlo radiative transfer code, to construct the 3D model of the radiation field of M 51, following the methodology defined in the DustPedia framework. In the interest of modelling, the assumed centre-to-centre distance separation between the two galaxies is similar to 10 kpc.Results. Our model is able to reproduce the global spectral energy distribution of the system, and it matches the resolved optical and infrared images fairly well. In total, 40.7% of the intrinsic stellar radiation of the combined system is absorbed by dust. Furthermore, we quantify the contribution of the various dust heating sources in the system, and find that the young stellar population of NGC 5194 is the predominant dust-heating agent, with a global heating fraction of 71.2%. Another 23% is provided by the older stellar population of the same galaxy, while the remaining 5.8% has its origin in NGC 5195. Locally, we find that the regions of NGC 5194 closer to NGC 5195 are significantly affected by the radiation field of the latter, with the absorbed energy fraction rising up to 38%. The contribution of NGC 5195 remains under the percentage level in the outskirts of the disc of NGC 5194. This is the first time that the heating of the diffuse dust by a companion galaxy is quantified in a nearby interacting system

The resolved scaling relations in DustPedia: Zooming in on the local Universe

We perform a homogeneous analysis of an unprecedented set of spatially resolved scaling relations (SRs) between ISM components and other properties in the range of scales 0.3-3.4 kpc. We also study some ratios: dust-to-stellar, dust-to-gas, and dust-to-metal. We use a sample of 18 large, spiral, face-on DustPedia galaxies. All the SRs are moderate/strong correlations except the dust-HI SR that does not exist or is weak for most galaxies. The SRs do not have a universal form but each galaxy is characterized by distinct correlations, affected by local processes and galaxy peculiarities. The SRs hold starting from 0.3 kpc, and if a breaking down scale exists it is < 0.3 kpc. By evaluating all galaxies at 3.4 kpc, differences due to peculiarities of individual galaxies are cancelled out and the corresponding SRs are consistent with those of whole galaxies. By comparing subgalactic and global scales, the most striking result emerges from the SRs involving ISM components: the dust-total gas SR is a good correlation at all scales, while the dust-H2 and dust-HI SRs are good correlations at subkpc/kpc and total scales, respectively. For the other explored SRs, there is a good agreement between small and global scales and this may support the picture where the main physical processes regulating the properties and evolution of galaxies occur locally. Our results are consistent with the hypothesis of self-regulation of the SF process. The analysis of subgalactic ratios shows that they are consistent with those derived for whole galaxies, from low to high z, supporting the idea that also these ratios could be set by local processes. Our results highlight the heterogeneity of galaxy properties and the importance of resolved studies on local galaxies in the context of galaxy evolution. They also provide observational constraints to theoretical models and updated references for high-z studies.Comment: 42 pages, 11 figures and 5 tables in the main text, 2 figures and 1 table in Appendix. Accepted for publication in A&

Survey of CF mutations in the clinical laboratory

BACKGROUND: Since it is impossible to sequence the complete CFTR gene routinely, clinical laboratories must rely on test systems that screen for a panel of the most frequent mutations causing disease in a high percentage of patients. Thus, in a cohort of 257 persons that were referred to our laboratory for analysis of CF gene mutations, reverse line probe assays for the most common CF mutations were performed. These techniques were evaluated as routine first-line analyses of the CFTR gene status. METHODS: DNA from whole blood specimens was extracted and subjected to PCR amplification of 9 exons and 6 introns of the CFTR gene. The resulting amplicons were hybridised to probes for CF mutations and polymorphisms, immobilised on membranes supplied by Roche Molecular Systems, Inc. and Innogenetics, Inc.. Denaturing gradient gel electrophoresis and sequencing of suspicious fragments indicating mutations were done with CF exon and intron specific primers. RESULTS: Of the 257 persons tested over the last three years (referrals based on 1) clinical symptoms typical for/indicative of CF, 2) indication for in vitro fertilisation, and 3) gene status determination because of anticipated parenthood and partners or relatives affected by CF), the reverse line blots detected heterozygote or homozygote mutations in the CFTR gene in 68 persons (26%). Eighty-three percent of those affected were heterozygous (47 persons) or homozygous (10 persons) for the ΔF508 allele. The only other CF-alleles that we found with these tests were the G542X allele (3 persons), the G551D allele (3 persons), the 3849+10kb C-T allele (2 persons) the R117H allele (2 persons) and the 621+1G-T allele (1 person). Of the fifteen IVS8-5T-polymorphisms detected in intron 8, seven (47%) were found in males referred to us from IVF clinics. These seven 5T-alleles were all coupled with a heterozygous ΔF508 allele, they make up 35% of the males with fertility problems (20 men) referred to us. CONCLUSIONS: In summary, the frequency of CF chromosomes in the cohort examined with these tests was 26%, with the ΔF508 allele affecting 83% of the CF chromosomes. It is a substantial improvement for routine CF diagnostics to have available a test system for 30 mutations plus the polypyrimidine length variants in intron 8. Our results show that this test system allows a routine first-line analyses of the CFTR gene status

Improved overall survival in patients with high-grade serous ovarian cancer is associated with CD16a+ immunologic neighborhoods containing NK cells, T cells and macrophages

BackgroundFor patients with high grade serous carcinoma of the ovary (HGSC), survival rates have remained static for the last half century. Despite the presence of tumor mutations and infiltration of immune cells, existing immunotherapies have achieved little success against HGSC. These observations highlight a gap in the understanding of how the immune system functions and interacts within HGSC tumors.MethodsWe analyzed duplicate core samples from 939 patients with HGSC to understand patterns of immune cell infiltration, localization, and associations with clinical features. We used high-parameter immunohistochemical/Opal multiplex, digital pathology, computational biology, and multivariate analysis to identify immune cell subsets and their associations with HGSC tumors.ResultsWe defined six patterns of cellular infiltration by spatially restricted unsupervised clustering of cell subsets. Each pattern was represented to some extent in most patient samples, but their specific distributions differed. Overall (OS) and progression-free survival (PFS) corresponded with higher infiltration of CD16a+ cells, and their co-localization with macrophages, T cells, NK cells, in one of six cellular neighborhoods that we defined with our spatial assessment.ConclusionsImmune cell neighborhoods containing CD16a+ cells are associated with improved OS and PFS for patients with HGSC. Patterns of immunologic neighborhoods differentiate patient outcomes, and could inform future, more precise approaches to treatment

Subcellular Location, Phosphorylation and Assembly into the Motor Complex of GAP45 during Plasmodium falciparum Schizont Development

An actomyosin motor complex assembled below the parasite's plasma membrane drives erythrocyte invasion by Plasmodium falciparum merozoites. The complex is comprised of several proteins including myosin (MyoA), myosin tail domain interacting protein (MTIP) and glideosome associated proteins (GAP) 45 and 50, and is anchored on the inner membrane complex (IMC), which underlies the plasmalemma. A ternary complex of MyoA, MTIP and GAP45 is formed that then associates with GAP50. We show that full length GAP45 labelled internally with GFP is assembled into the motor complex and transported to the developing IMC in early schizogony, where it accumulates during intracellular development until merozoite release. We show that GAP45 is phosphorylated by calcium dependent protein kinase 1 (CDPK1), and identify the modified serine residues. Replacing these serine residues with alanine or aspartate has no apparent effect on GAP45 assembly into the motor protein complex or its subcellular location in the parasite. The early assembly of the motor complex suggests that it has functions in addition to its role in erythrocyte invasion

Plasmodium gametocytes display homing and vascular transmigration in the host bone marrow

Transmission of Plasmodium parasites to the mosquito requires the formation and development of gametocytes. Studies in infected humans have shown that only the most mature forms of Plasmodium falciparum gametocytes are present in circulation, whereas immature forms accumulate in the hematopoietic environment of the bone marrow. We used the rodent model Plasmodium berghei to study gametocyte behavior through time under physiological conditions. Intravital microscopy demonstrated preferential homing of early gametocyte forms across the intact vascular barrier of the bone marrow and the spleen early during infection and subsequent development in the extravascular environment. During the acute phase of infection, we observed vascular leakage resulting in further parasite accumulation in this environment. Mature gametocytes showed high deformability and were found entering and exiting the intact vascular barrier. We suggest that extravascular gametocyte localization and mobility are essential for gametocytogenesis and transmission of Plasmodium to the mosquito

Exploring the interstellar medium of NGC 891 at millimeter wavelengths using the NIKA2 camera

In the framework of the IMEGIN Large Program, we used the NIKA2 camera on the IRAM 30-m telescope to observe the edge-on galaxy NGC 891 at 1.15 mm and 2 mm and at a FWHM of 11.1" and 17.6", respectively. Multiwavelength data enriched with the new NIKA2 observations fitted by the HerBIE SED code (coupled with the THEMIS dust model) were used to constrain the physical properties of the ISM. Emission originating from the diffuse dust disk is detected at all wavelengths from mid-IR to mm, while mid-IR observations reveal warm dust emission from compact HII regions. Indications of mm excess emission have also been found in the outer parts of the galactic disk. Furthermore, our SED fitting analysis constrained the mass fraction of the small (< 15 Angstrom) dust grains. We found that small grains constitute 9.5% of the total dust mass in the galactic plane, but this fraction increases up to ~ 20% at large distances (|z| > 3 kpc) from the galactic plane.Comment: To appear in Proc. of the mm Universe 2023 conference, Grenoble (France), June 2023, published by F. Mayet et al. (Eds), EPJ Web of conferences, EDP Science

IAS/CEA Evolution of Dust in Nearby Galaxies (ICED): the spatially-resolved dust properties of NGC4254

We present the first preliminary results of the project \textit{ICED}, focusing on the face-on galaxy NGC4254. We use the millimetre maps observed with NIKA2 at IRAM-30m, as part of the IMEGIN Guaranteed Time Large Program, and of a wide collection of ancillary data (multi-wavelength photometry and gas phase spectral lines) that are publicly available. We derive the global and local properties of interstellar dust grains through infrared-to-radio spectral energy distribution fitting, using the hierarchical Bayesian code HerBIE, which includes the grain properties of the state-of-the-art dust model, THEMIS. Our method allows us to get the following dust parameters: dust mass, average interstellar radiation field, and fraction of small grains. Also, it is effective in retrieving the intrinsic correlations between dust parameters and interstellar medium properties. We find an evident anti-correlation between the interstellar radiation field and the fraction of small grains in the centre of NGC4254, meaning that, at strong radiation field intensities, very small amorphous carbon grains are efficiently destroyed by the ultra-violet photons coming from newly formed stars, through photo-desorption and sublimation. We observe a flattening of the anti-correlation at larger radial distances, which may be driven by the steep metallicity gradient measured in NGC4254.Comment: to appear in Proc. of the mm Universe 2023 conference, Grenoble (France), June 2023, published by F. Mayet et al. (Eds), EPJ Web of conferences, EDP Science