An ancestral secretory apparatus in the protozoan parasite Giardia intestinalis

The protozoan parasite Giardia intestinalis belongs to one of the earliest diverged eukaryotic lineages. This is also reflected in a simple intracellular organization, as Giardia lacks common subcellular compartments such as mitochondria, peroxisomes, and apparently also a Golgi apparatus. During encystation, developmentally regulated formation of large secretory compartments containing cyst wall material occurs. Despite the lack of any morphological similarities, these encystation-specific vesicles (ESVs) show several biochemical characteristics of maturing Golgi cisternae. Previous studies suggested that Golgi structure and function are induced only during encystation in Giardia, giving rise to the hypothesis that ESVs, as a Giardia Golgi equivalent, are generated de novo. Alternatively, ESV compartments could be built on the template structure of a cryptic Golgi in trophozoites in response to ER export of cyst wall material during encystation. We addressed this question by defining the molecular framework of the Giardia secretory apparatus using a comparative genomic approach. Analysis of the corresponding transcriptome during growth and encystation revealed surprisingly little stage-specific regulation. A panel of antibodies was generated against selected marker proteins to investigate the developmental dynamics of the endomembrane system. We show evidence that Giardia accommodates the export of large amounts of cyst wall material through re-organization of membrane compartment(s) in trophozoites with biochemical similarities to ESVs. This suggests that ESVs are selectively stabilized Golgi-like compartments in a unique and archetypical secretory system, which arise from a structural template in trophozoites rather than being generated de novo

The 1986?1989 ENSO cycle in a chemical climate model

International audienceA pronounced ENSO cycle occurred from 1986 to 1989, accompanied by distinct dynamical and chemical anomalies in the global troposphere and stratosphere. Reproducing these effects with current climate models not only provides a model test but also contributes to our still limited understanding of ENSO's effect on stratosphere-troposphere coupling. We performed several sets of ensemble simulations with a chemical climate model (SOCOL) forced with global sea surface temperatures. Results were compared with observations and with large-ensemble simulations performed with an atmospheric general circulation model (MRF9). We focus our analysis on the extratropical stratosphere and its coupling with the troposphere. In this context, the circulation over the North Atlantic sector is particularly important. Observed differences between the El Niño winter 1987 and the La Niña winter 1989 include a negative North Atlantic Oscillation index with corresponding changes in temperature and precipitation patterns, a weak polar vortex, a warm Arctic middle stratosphere, negative and positive total ozone anomalies in the tropics and at middle to high latitudes, respectively, as well as anomalous upward and poleward Eliassen-Palm (EP) flux in the midlatitude lower stratosphere. Most of the tropospheric features are well reproduced in the ensemble means in both models, though the amplitudes are underestimated. In the stratosphere, the SOCOL simulations compare well with observations with respect to zonal wind, temperature, EP flux, and ozone, but magnitudes are underestimated in the middle stratosphere. The polar vortex strength is well reproduced, but within-ensemble variability is too large for obtaining a significant signal in Arctic temperature and ozone. With respect to the mechanisms relating ENSO to stratospheric circulation, the results suggest that both, upward and poleward components of anomalous EP flux are important for obtaining the stratospheric signal and that an increase in strength of the Brewer-Dobson circulation is part of that signal

Cyst-Wall-Protein-1 is fundamental for Golgi-like organelle neogenesis and cyst-wall biosynthesis in Giardia lamblia

The genome of the protozoan parasite Giardia lamblia is organized in two diploid nuclei, which has so far precluded complete analysis of gene function. Here we use a previously developed Cre/loxP-based knock-out and selection marker salvage strategy in the human-derived isolate WB-C6 to eliminate all four copies of the Cyst-Wall-Protein-1 locus (CWP1). Because these loci are silenced in proliferating trophozoites and highly expressed only in encysting cells, CWP1 ablation allows functional characterization of a conditional phenotype in parasites induced to encyst. We show that encysting Δcwp1 cells are unable to establish the stage-regulated trafficking machinery with Golgi-like encystation-specific vesicles required for cyst-wall formation but show morphological hallmarks of cyst development and karyokinesis. This ‘pseudocyst’ phenotype is rescued by transfection of Δcwp1 cells with an episomally maintained CWP1 expression vector. Genome editing in genera Giardia and Trypanosoma are the only reported examples addressing questions on pathogen transmission within the Excavata supergroup

Transfer of Anti-Rotavirus Antibodies during Pregnancy and in Milk Following Maternal Vaccination with a Herpes Simplex Virus Type-1 Amplicon Vector.

Rotaviruses (RVs) are important enteric pathogens of newborn humans and animals, causing diarrhea and in rare cases death, especially in very young individuals. Rotavirus vaccines presently used are modified live vaccines that lack complete biological safety. Previous work from our laboratory suggested that vaccines based on in situ produced, non-infectious rotavirus-like particles (RVLPs) are efficient while being entirely safe. However, using either vaccine, active mucosal immunization cannot induce protective immunity in newborns due to their immature immune system. We therefore hypothesized that offspring from vaccinated dams are passively immunized either by transfer of maternal antibodies during pregnancy or by taking up antibodies from milk. Using a codon optimized polycistronic gene expression cassette packaged into herpesvirus particles, the simultaneous expression of the RV capsid genes led to the intracellular formation of RVLPs in various cell lines. Vaccinated dams developed a strong RV specific IgG antibody response determined in sera and milk of both mother and pups. Moreover, sera of naïve pups nursed by vaccinated dams also had RV specific antibodies suggesting a lactogenic transfer of antibodies. Although full protection of pups was not achieved in this mouse model, our observations are important for the development of improved vaccines against RV in humans as well as in various animal species

Technical Note: Chemistry-climate model SOCOL: version 2.0 with improved transport and chemistry/microphysics schemes

International audienceWe describe version 2.0 of the chemistry-climate model (CCM) SOCOL. The new version includes fundamental changes of the transport scheme such as transporting all chemical species of the model individually and applying a family-based correction scheme for mass conservation for species of the nitrogen, chlorine and bromine groups, a revised transport scheme for ozone, furthermore more detailed halogen reaction and deposition schemes, and a new cirrus parameterisation in the tropical tropopause region. By means of these changes the model manages to overcome or considerably reduce deficiencies recently identified in SOCOL version 1.1 within the CCM Validation activity of SPARC (CCMVal). In particular, as a consequence of these changes, regional mass loss or accumulation artificially caused by the semi-Lagrangian transport scheme can be significantly reduced, leading to much more realistic distributions of the modelled chemical species, most notably of the halogens and ozone

Identification of a small molecule that compromises the structural integrity of viroplasms and rotavirus double-layered particles

Despite the availability of two attenuated vaccines, rotavirus (RV) gastroenteritis remains an important cause of mortality among children in developing countries, causing about 215,000 infant deaths annually. Currently, there are no specific antiviral therapies available. RV is a nonenveloped virus with a segmented double-stranded RNA genome. Viral genome replication and assembly of transcriptionally active double-layered particles (DLPs) take place in cytoplasmic viral structures called viroplasms. In this study, we describe strong impairment of the early stages of RV replication induced by a small molecule known as an RNA polymerase III inhibitor, ML-60218 (ML). This compound was found to disrupt already assembled viroplasms and to hamper the formation of new ones without the need for de novo transcription of cellular RNAs. This phenotype was correlated with a reduction in accumulated viral proteins and newly made viral genome segments, disappearance of the hyperphosphorylated isoforms of the viroplasm-resident protein NSP5, and inhibition of infectious progeny virus production. In in vitro transcription assays with purified DLPs, ML showed dose-dependent inhibitory activity, indicating the viral nature of its target. ML was found to interfere with the formation of higher-order structures of VP6, the protein forming the DLP outer layer, without compromising its ability to trimerize. Electron microscopy of ML-treated DLPs showed dose-dependent structural damage. Our data suggest that interactions between VP6 trimers are essential, not only for DLP stability, but also for the structural integrity of viroplasms in infected cells

Comparing Atmospheric Correction Performance for Sentinel-2 and Landsat-8 data

In terms of atmospheric impact, the volcanic eruption of Mt. Pinatubo (1991) is thebest characterized large eruption on record. We investigate here the stratosphericwarming following the Pinatubo eruption derived from SAGE II extinction data includ-ing most recent improvements in the processing algorithm and a data filling procedure in the opacity-induced “gap” regions. From these data, which cover wavelengths of1.024 micrometer and shorter, we derived aerosol size distributions which properly re-produce extinction coefficients at much longer wavelength. This provides a good basisfor calculating the absorption of terrestrial infrared radiation and the resulting strato-spheric heating. However, we also show that the use of this dataset in the global chemistry-climate model (CCM) SOCOL leads to exaggerated aerosol-induced strato-spheric heating compared to observations, even partly larger than the already too highvalues found by many models in recent general circulation model (GCM) and CCMintercomparisons. This suggests that the overestimation of the stratospheric warm-ing after the Pinatubo eruption arises from deficiencies in the model radiation codes rather than an insufficient observational data basis. Conversely, our approach reducesthe infrared absorption in the tropical tropopause region, in better agreement with thepost-volcanic temperature record at these altitudes.ISSN:1680-7375ISSN:1680-736

Bax Function in the Absence of Mitochondria in the Primitive Protozoan Giardia lamblia

Bax-induced permeabilization of the mitochondrial outer membrane and release of cytochrome c are key events in apoptosis. Although Bax can compromise mitochondria in primitive unicellular organisms that lack a classical apoptotic machinery, it is still unclear if Bax alone is sufficient for this, or whether additional mitochondrial components are required. The protozoan parasite Giardia lamblia is one of the earliest branching eukaryotes and harbors highly degenerated mitochondrial remnant organelles (mitosomes) that lack a genome. Here we tested whether human Bax expressed in Giardia can be used to ablate mitosomes. We demonstrate that these organelles are neither targeted, nor compromised, by Bax. However, specialized compartments of the regulated secretory pathway are completely ablated by Bax. As a consequence, maturing cyst wall proteins that are sorted into these organelles are released into the cytoplasm, causing a developmental arrest and cell death. Interestingly, this ectopic cargo release is dependent on the carboxy-terminal 22 amino acids of Bax, and can be prevented by the Bax-inhibiting peptide Ku70. A C-terminally truncated Bax variant still localizes to secretory organelles, but is unable to permeabilize these membranes, uncoupling membrane targeting and cargo release. Even though mitosomes are too diverged to be recognized by Bax, off-target membrane permeabilization appears to be conserved and leads to cell death completely independently of mitochondria

Diurnal changes in middle atmospheric H2O and O3: Observations in the Alpine region and climate models

International audienceIn this paper we investigate daily variations in middle atmospheric water vapor and ozone based on data from two ground-based microwave radiometers located in the Alpine region of Europe. Temperature data are obtained from a lidar located near the two stations and from the SABER experiment on the TIMED satellite. This unique set of observations is complemented by three different three-dimensional (3-D) chemistry-climate models (Monitoring of Stratospheric Depletion of the Ozone Layer (MSDOL), Laboratoire de Météorologie Dynamique Reactive Processes Ruling the Ozone Budget in the Stratosphere (LMDz-REPROBUS), and Solar Climate Ozone Links (SOCOL)) and the 2-D atmospheric global-scale wave model (GSWM). The first part of the paper is focused on the first Climate and Weather of the Sun-Earth System (CAWSES) tidal campaign that consisted of a period of intensive measurements during September 2005. Variations in stratospheric water vapor are found to be in the order of 1% depending on altitude. Meridional advection of tidal nature is likely to be the dominant driving factor throughout the whole stratosphere, while vertical advection becomes more important in the mesosphere. Observed ozone variations in the upper stratosphere and lower mesosphere show amplitudes of several percent in accordance with photochemical models. Variations in lower stratospheric ozone are not solely governed by photochemistry but also by dynamics, with the temperature dependence of the photochemistry becoming more important. The second part presents an investigation of the seasonal dependence of daily variations. Models tend to underestimate the H2O diurnal amplitudes, especially during summer in the upper stratosphere. Good agreement between models and observations is found for ozone in the upper stratosphere, which reflects the fact that the O3 daily variations are driven by the photochemistry that is well modeled

Evaluation of the inter-annual variability of stratospheric chemical composition in chemistry-climate models using ground-based multi species time series

The variability of stratospheric chemical composition occurs on a broad spectrum of timescales, ranging from day to decades. A large part of the variability appears to be driven by external forcings such as volcanic aerosols, solar activity, halogen loading, levels of greenhouse gases (GHG), and modes of climate variability (quasi-biennial oscillation (QBO), El Niño-Southern Oscillation (ENSO)). We estimate the contributions of different external forcings to the interannual variability of stratospheric chemical composition and evaluate how well 3-D chemistry-climate models (CCMs) can reproduce the observed response-forcing relationships. We carry out multivariate regression analyses on long time series of observed and simulated time series of several traces gases in order to estimate the contributions of individual forcings and unforced variability to their internannual variability. The observations are typically decadal time series of ground-based data from the international Network for the Detection of Atmospheric Composition Change (NDACC) and the CCM simulations are taken from the CCMVal-2 REF-B1 simulations database. The chemical species considered are column O3, HCl, NO2, and N2O. We check the consistency between observations and model simulations in terms of the forced and internal components of the total interannual variability (externally forced variability and internal variability) and identify the driving factors in the interannual variations of stratospheric chemical composition over NDACC measurement sites. Overall, there is a reasonably good agreement between regression results from models and observations regarding the externally forced interannual variability. A much larger fraction of the observed and modelled interannual variability is explained by external forcings in the tropics than in the extratropics, notably in polar regions. CCMs are able to reproduce the amplitudes of responses in chemical composition to specific external forcings. However, CCMs tend to underestimate very substantially the internal variability and hence the total interannual variability for almost all species considered. This lack of internal variability in CCMs might partly originate from the surface forcing of these CCMs by analysed SSTs. The results illustrate the potential of NDACC ground-based observations for evaluating CCMs