STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena

The base of the cilium comprising the transition zone (TZ) and transition fibers (TF) acts as a selecting gate to regulate the intraflagellar transport (IFT)-dependent trafficking of proteins to and from cilia. Before entering the ciliary compartment, IFT complexes and transported cargoes accumulate at or near the base of the cilium. The spatial organization of IFT proteins at the cilia base is key for understanding cilia formation and function. Using stochastic optical reconstruction microscopy (STORM) and computational averaging, we show that seven TZ, nine IFT, three Bardet–Biedl syndrome (BBS), and one centrosomal protein, form 9-clustered rings at the cilium base of a ciliate Tetrahymena thermophila. In the axial dimension, analyzed TZ proteins localize to a narrow region of about 30 nm while IFT proteins dock approximately 80 nm proximal to TZ. Moreover, the IFT-A subcomplex is positioned peripheral to the IFT-B subcomplex and the investigated BBS proteins localize near the ciliary membrane. The positioning of the HA-tagged N- and C-termini of the selected proteins enabled the prediction of the spatial orientation of protein particles and likely cargo interaction sites. Based on the obtained data, we built a comprehensive 3D-model showing the arrangement of the investigated ciliary proteins

Current status of MELCOR 2.2 for fusion safety analyses

MELCOR is an integral code developed by Sandia National Laboratories (SNL) for the US Nuclear Regulatory Commission (USNRC) to perform severe accident analyses of Light Water Reactors (LWR). More recently, MELCOR capabilities are being extended also to analyze non-LWR fission technologies. Within the European MELCOR User Group (EMUG), organized in the framework of USNRC Cooperative Severe Accident Research Program (CSARP), an activity on the evaluation of the applicability of MELCOR 2.2 for fusion safety analyses has been launched and it has been coordinated by ENEA. The aim of the activity was to identify the physical models to be possibly implemented in MELCOR 2.2 necessary for fusion safety analyses, and to check if those models are already available in MELCOR 1.8.6 for fusion version, developed by Idaho National Laboratory (INL). From this activity, a list of modeling needs emerged from the safety analyses of fusion-related installations have been identified and described. Then, the importance of the various needs, intended as the priority for model implementation in the MELCOR 2.2 code, has been evaluated according to the technical expert judgement of the authors. In the present paper, the identified modeling needs are discussed. The ultimate goal would be to propose to have a single integrated MELCOR 2.2 code release capable to cover both fission and fusion applications

Current status of Melcor 2.2 for fusion safety analyses

MELCOR is an integral code developed by Sandia National Laboratories (SNL) for the US Nuclear Regulatory Commission (USNRC) to perform severe accident analyses of Light Water Reactors (LWR). More recently, MELCOR capabilities are being extended also to analyze non-LWR fission technologies. Within the European MELCOR User Group (EMUG), organized in the framework of the USNRC Cooperative Severe Accident Research Program (CSARP), an activity on the evaluation of the applicability of MELCOR 2.2 for fusion safety analyses has been launched and it has been coordinated by ENEA. The aim of the activity was to identify the physical models to be possibly implemented in MELCOR 2.2 necessary for fusion safety analyses, and to check if those models are already available in MELCOR 1.8.6 fusion version, developed by Idaho National Laboratory (INL). From this activity, a list of modeling needs that emerged from the safety analyses of fusion-related installations has been identified and described. Then, the importance of the various needs, intended as the priority for model implementation in the MELCOR 2.2 code, has been evaluated according to the technical expert judgment of the authors. In the present paper, the identified modeling needs are discussed. The ultimate goal would be to propose to have a single integrated MELCOR 2.2 code release capable to cover both fission and fusion applications

The Role of Mislocalized Phototransduction in Photoreceptor Cell Death of Retinitis Pigmentosa

Most of inherited retinal diseases such as retinitis pigmentosa (RP) cause photoreceptor cell death resulting in blindness. RP is a large family of diseases in which the photoreceptor cell death can be caused by a number of pathways. Among them, light exposure has been reported to induce photoreceptor cell death. However, the detailed mechanism by which photoreceptor cell death is caused by light exposure is unclear. In this study, we have shown that even a mild light exposure can induce ectopic phototransduction and result in the acceleration of rod photoreceptor cell death in some vertebrate models. In ovl, a zebrafish model of outer segment deficiency, photoreceptor cell death is associated with light exposure. The ovl larvae show ectopic accumulation of rhodopsin and knockdown of ectopic rhodopsin and transducin rescue rod photoreceptor cell death. However, knockdown of phosphodiesterase, the enzyme that mediates the next step of phototransduction, does not. So, ectopic phototransduction activated by light exposure, which leads to rod photoreceptor cell death, is through the action of transducin. Furthermore, we have demonstrated that forced activation of adenylyl cyclase in the inner segment leads to rod photoreceptor cell death. For further confirmation, we have also generated a transgenic fish which possesses a human rhodopsin mutation, Q344X. This fish and rd10 model mice show photoreceptor cell death caused by adenylyl cyclase. In short, our study indicates that in some RP, adenylyl cyclase is involved in photoreceptor cell death pathway; its inhibition is potentially a logical approach for a novel RP therapy

Main outcomes of the Phebus FPT1 uncertainty and sensitivity analysis in the EU-MUSA project

The Management and Uncertainties of Severe Accidents (MUSA) project was funded in HORIZON 2020 and is coordinated by CIEMAT (Spain). The project aims at consolidating a harmonized approach for the analysis of uncertainties and sensitivities associated with Severe Accidents (SAs) analysis, focusing on source term figures of merit. The Application of Uncertainty Quantification (UQ) Methods against Integral Experiments (AUQMIE – Work Package 4 (WP4)), led by ENEA (Italy), was devoted to apply and test UQ methodologies adopting the internationally recognized PHEBUS FPT1 test. FPT1 was chosen to test UQ methodologies because, even though it is a simplified SA scenario, it was representative of the in-vessel phase of a severe accident initiated by a break in the cold leg of a PWR primary circuit. WP4 served as a platform to identify and discuss the issues encountered in the application of UQ methodol ogies to SA analyses (e.g. discuss the UQ methodology, perform the coupling between the SA codes and the UQ tools, define the results post-processing methods, etc.). The purpose of this paper is to describe the MUSA PHEBUS FPT1 uncertainty application exercise with the related specifications and the methodologies used by the partners to perform the UQ exercise. The main outcomes and lessons learned of the analysis are: scripting was in general needed for the SA code and uncertainty tool coupling and to have more flexibility; particular attention should be devoted to the proper choice of the input uncertain parameters; outlier values of figures of merit should be carefully analyzed; the computational time is a key element to perform UQ in SA; the large number of uncertain input parameters may complicate the interpretation of correlation or sensitivity analysis; there is the need for a statistically solid handling of failed calculations

The Role of Glypicans in Wnt Inhibitory Factor-1 Activity and the Structural Basis of Wif1's Effects on Wnt and Hedgehog Signaling

Proper assignment of cellular fates relies on correct interpretation of Wnt and Hedgehog (Hh) signals. Members of the Wnt Inhibitory Factor-1 (WIF1) family are secreted modulators of these extracellular signaling pathways. Vertebrate WIF1 binds Wnts and inhibits their signaling, but its Drosophila melanogaster ortholog Shifted (Shf) binds Hh and extends the range of Hh activity in the developing D. melanogaster wing. Shf activity is thought to depend on reinforcing interactions between Hh and glypican HSPGs. Using zebrafish embryos and the heterologous system provided by D. melanogaster wing, we report on the contribution of glypican HSPGs to the Wnt-inhibiting activity of zebrafish Wif1 and on the protein domains responsible for the differences in Wif1 and Shf specificity. We show that Wif1 strengthens interactions between Wnt and glypicans, modulating the biphasic action of glypicans towards Wnt inhibition; conversely, glypicans and the glypican-binding “EGF-like” domains of Wif1 are required for Wif1's full Wnt-inhibiting activity. Chimeric constructs between Wif1 and Shf were used to investigate their specificities for Wnt and Hh signaling. Full Wnt inhibition required the “WIF” domain of Wif1, and the HSPG-binding EGF-like domains of either Wif1 or Shf. Full promotion of Hh signaling requires both the EGF-like domains of Shf and the WIF domains of either Wif1 or Shf. That the Wif1 WIF domain can increase the Hh promoting activity of Shf's EGF domains suggests it is capable of interacting with Hh. In fact, full-length Wif1 affected distribution and signaling of Hh in D. melanogaster, albeit weakly, suggesting a possible role for Wif1 as a modulator of vertebrate Hh signaling

Wild vascular plants gathered for consumption in the Polish countryside: a review

BACKGROUND: This paper is an ethnobotanical review of wild edible plants gathered for consumption from the end of the 18(th )century to the present day, within the present borders of Poland. METHODS: 42 ethnographic and botanical sources documenting the culinary use of wild plants were analyzed. RESULTS: The use of 112 species (3.7% of the flora) has been recorded. Only half of them have been used since the 1960s. Three species: Cirsium rivulare, Euphorbia peplus and Scirpus sylvaticus have never before been reported as edible by ethnobotanical literature. The list of wild edible plants which are still commonly gathered includes only two green vegetables (Rumex acetosa leaves for soups and Oxalis acetosella as children's snack), 15 folk species of fruits and seeds (Crataegus spp., Corylus avellana, Fagus sylvatica, Fragaria vesca, Malus domestica, Prunus spinosa, Pyrus spp., Rosa canina, Rubus idaeus, Rubus sect. Rubus, Sambucus nigra, Vaccinium myrtillus, V. oxycoccos, V. uliginosum, V. vitis-idaea) and four taxa used for seasoning or as preservatives (Armoracia rusticana root and leaves, Carum carvi seeds, Juniperus communis pseudo-fruits and Quercus spp. leaves). The use of other species is either forgotten or very rare. In the past, several species were used for food in times of scarcity, most commonly Chenopodium album, Urtica dioica, U. urens, Elymus repens, Oxalis acetosella and Cirsium spp., but now the use of wild plants is mainly restricted to raw consumption or making juices, jams, wines and other preserves. The history of the gradual disappearance of the original barszcz, Heracleum sphondylium soup, from Polish cuisine has been researched in detail and two, previously unpublished, instances of its use in the 20(th )century have been found in the Carpathians. An increase in the culinary use of some wild plants due to media publications can be observed. CONCLUSION: Poland can be characterized as a country where the traditions of culinary use of wild plants became impoverished very early, compared to some parts of southern Europe. The present use of wild plants, even among the oldest generation, has been almost entirely restricted to fruits

Dynamic Coupling of Pattern Formation and Morphogenesis in the Developing Vertebrate Retina

In this Research Article, Picker et al. show how cells in the retina get their spatial coordinates