Cell Response to the Exposure to Chitosan-TPP//Alginate Nanogels.

Hydrophilic nanocarriers formed by electrostatic interaction of chitosan with oppositely charged macromolecules have a high potential as vectors in biomedical and pharmaceutical applications. However, comprehensive information about the fate of such nanomaterials in biological environment is lacking. We used chitosan from both animal and fungal sources to form well-characterized chitosan-pentasodium triphosphate (TPP)//alginate nanogels suitable for comparative studies. Upon exposure of human colon cancer cells (HT29 and CaCo2), breast cancer cells (MDA-MB-231 and MCF-7), glioblastoma cells (LN229), lung cancer cells (A549), and brain-derived endothelial cells (HCEC) to chitosan-(TPP)//alginate nanogels, cell type-, nanogel dosage-, and exposure time-dependent responses are observed. Comparing chitosan-TPP//alginate nanogels prepared from either animal or fungal source in terms of nanogel formation, cell uptake, reactive oxygen species production, and metabolic cell activity, no significant differences become obvious. The results identify fungal chitosan as an alternative to animal chitosan in particular if biomedical/pharmaceutical applications are intended.

A Miocene mud volcano and its plumbing system: A chaotic complex revisited (Monferrato, NW Italy)

Chaotic deposits are frequently reported in the geological literature and are commonly interpreted as olistostromes or tectonic melanges. A chaotic complex in the Cenozoic succession of Monferrato (NW Italy) consists of interbedded mud breccia and burrowed silty clays that are pierced by sheared mud breccias and embed carbonate-cemented blocks. These may be represented by microcrystalline limestones or strongly cemented matrix-supported breccias locally containing remains of chemosymbiotic organisms (lucinid bivalves). Moreover, cylindrical concretions, up to 15 cm in diameter and 1 m long, occur in the chaotic complex and crosscut bedding planes at high angles. The cement of all these lithified portions is mainly dolomite characterized by low delta(13)C values (from -10.3 to -23parts per thousand PDB) and delta(18)O values up to + 7parts per thousand PDB. The delta(13)C values testify to precipitation of carbonates induced by microbial oxidation of methane, whereas the markedly positive delta(18)C signature, ubiquitous in the cylindrical concretions, is the evidence for the presence and destabilization of gas hydrates. The studied section provides a well-exposed example of the geological record of the birth, life, and death of a mud volcano. Unsheared, soft mud breccias represent mud flows along the flanks of the volcano, whereas sheared mud breccias are the result of the injection of unconsolidated overpressured fine-grained sediments, both taking place during ``eruptive'' phases. They were followed by more quiet stages of hemipelagic sedimentation, burrowing, and CH4 seeping. The cylindrical concretions represent the first described ancient example of the chimneys observed in present-day mud-volcano settings. They are the remnants of a cold-seep plumbing network that crosscut the mud volcano edifice. The chimneys were the pathway for the expulsion toward the sea floor of gas- and sediment-charged fluids likely originated from destabilization of methane gas hydrates. The association of mud breccias and methane-derived carbonates may not be due to mass gravity flows but can be primary and, therefore, is a diagnostic criterion for recognizing chaotic deposits due to mud volcano activity in the geological record.

Electromagnetic radiation initiated by hadronic jets from microquasars in the ISM

Microquasars are potential candidates to produce a non-negligible fraction of the observed galactic cosmic rays. The protons accelerated at the jet termination shock interact with the interstellar medium and may produce detectable fluxes of extended emission at different energy bands: high-energy and very high-energy gamma-rays produced by neutral pion-decay, synchrotron and bremsstrahlung emission in a wide energy range generated by the secondary electrons produced by charged pion-decay. We discuss the association between this scenario and some of the unidentified EGRET sources in the galactic plane.

Factors promoting the exposure to bioaerosols among crop workers

Introduction. Agricultural workers are among the professional groups most at risk of developing acute or chronic respiratory problems. Despite this fact, the etiology of these occupational diseases is poorly known, even in important sectors of agriculture such as the crops sector. Cereals can be colonized by a large number of fungal species throughout the plants' growth, but also during grain storage. Some of these fungi deliver toxins that can have a serious impact on human health when they are ingested via wheat products. Although International and European legislation on contaminants in food, including mycotoxins, include measures to ensure protection of public health by setting down the maximum levels for certain contaminants, the risks associated with the inhalation of such molecules during grain handling remains poorly documented. Goal of study. This project's objective was to characterize worker exposure to pathogenic, irritative or allergenic microorganisms and to identify the abiotic or biotic factors that reduce the growth of these microorganisms in crops. Indeed, the proliferation of microorganisms on wheat is dependent on temperature, rainfall and human disturbance (e.g. usage of tillage, addition of fungicides). A change in the concentration of these microorganisms in the substrate will directly result in a change in the concentration of aerosolized particles of the same microorganisms. Therefore, the exposure of worker to bioaérosols will also change. The Vaud region of Switzerland is a perfect region for conduct such a project as weather conditions vary and agricultural land management programs are divers at a small geographic scale. Methods. Bioaerosols and wheat dust have been sampled during wheat harvesting of summer 2010 at 100 sites uniformly distributed in the Vaud region that are representative of the different agriculture practices. Personal exposure has been evaluated for different wheat related activities: harvesting, grain unload, baling straw, the cleaning of harvesters and silos. Aerosols have been sampled at a rate of 2L/min between 15 min to 4 hours (t) on a 5m PVC filter for estimating the total dust inhaled, on gelatine filter for the identification and quantification of molds, and on a 0.45um polycarbonate filter for endotoxin quantification. Altitude, temperature and annual average rainfall were considered for each site. The physical and chemical characteristics of soils were determined using the methods in effect at Sol Council (Nyon). Total dust has been quantified following NIOSH 0500 method. Reactive endotoxine activity has been determined with Limulus Amebocyte Lysate Assay. All molds have been identified by the pyrosequencing of ITS2 amplicons generated from bioaerosol or wheat dust genomic DNA. Results & Conclusions. Our results confirm the previous quantitative data on the worker exposure to wheat dust. In addition, they show that crop workers are systematically exposed to complex mixtures of allergens, irritants or cytotoxic components. The novelty of our study is the systematic detection of molds such as Fusarium - that is a mycotoxins producer - in the bioaerosols. The results are interpreted by taking in account the agriculture practice, the Phosphorus : Carbon : Nitrogen ratio of the soil, the altitude and the average of rainy days per year.

Leptonic secondary emission in a hadronic microquasar model

Context.It has been proposed that the origin of the very high-energy photons emitted from high-mass X-ray binaries with jet-like features, so-called microquasars (MQs), is related to hadronic interactions between relativistic protons in the jet and cold protons of the stellar wind. Leptonic secondary emission should be calculated in a complete hadronic model that includes the effects of pairs from charged pion decays inside the jets and the emission from pairs generated by gamma-ray absorption in the photosphere of the system. Aims.We aim at predicting the broadband spectrum from a general hadronic microquasar model, taking into account the emission from secondaries created by charged pion decay inside the jet. Methods.The particle energy distribution for secondary leptons injected along the jets is consistently derived taking the energy losses into account. The spectral energy distribution resulting from these leptons is calculated after assuming different values of the magnetic field inside the jets. We also compute the spectrum of the gamma-rays produced by neutral pion-decay and processed by electromagnetic cascades under the stellar photon field. Results.We show that the secondary emission can dominate the spectral energy distribution at low energies (~1 MeV). At high energies, the production spectrum can be significantly distorted by the effect of electromagnetic cascades. These effects are phase-dependent, and some variability modulated by the orbital period is predicted.

Formaldehyde-fixation of platelets for flow cytometric measurement of phosphatidylserine exposure is feasible.

Strong platelet activation results in a redistribution of negatively charged phospholipids from the cytosolic to the outer leaflet of the cellular membrane. Annexin V has a high affinity to negatively charged phospholipids and can be used to identify procoagulant platelets. Formaldehyde fixation can cause factitious Annexin V binding. Our aim was to evaluate a method for fixing platelets avoiding additional Annexin V binding. We induced expression of negatively charged phospholipids on the surface of a fraction of platelets by combined activation with convulxin and thrombin in the presence of Annexin V-fluorescein isothiocyanate and calcium. Aliquots of resting and activated platelets were fixed with a low concentration, calcium-free formaldehyde solution. Both native platelets and fixed platelets were analyzed by flow cytometry immediately and after a 24-h storage at 4°C. We observed that the percentage of Annexin V positive resting platelets ranged from 1.5 to 9.3% for the native samples and from 0.4 to 12.8% for the fixed samples (P=0.706, paired t-test). The amount of Annexin V positive convulxin/thrombin activated platelets varied from 12.9 to 35.4% without fixation and from 15.3 to 36.3% after formalin fixation (P=0.450). After a 24-h storage at 4°C, Annexin V positive platelets significantly increased both in the resting and in the convulxin/thrombin activated samples of native platelets (both P<0.001), while results for formalin fixed platelets did not differ from baseline values (P=0.318 for resting fixed platelets; P=0.673 for activated fixed platelets). We conclude that platelet fixation with a low concentration, calcium-free formaldehyde solution does not alter the proportion of Annexin V positive platelets. This method can be used to investigate properties of procoagulant platelets by multicolor flow-cytometric analysis requiring fixation steps.

Risques microbiologiques dans les espaces fermés : exemple d'un centre de compostage et de cultures en serres

La manipulation de végétaux ou de débris végétaux dans des espaces fermés (hangar pour le compostage, serres pour les cultures) est fréquemment effectuée en France. Néanmoins, le niveau d'exposition des travailleurs des différentes filières concernées aux bioaérosols dégagés par ces substrats, est peu connu. Dans cette note, deux études ont été choisies pour illustrer l'ampleur de ce type d'exposition dans des espaces fermés. La première publication concerne un centre de compostage et la deuxième publication cible des serres de culture de la tomate et du concombre. L'extrapolation des résultats obtenus à partir de prélèvements d'ambiance à l'exposition professionnelle des travailleurs est tout particulièrement discutée.

Activation mechanisms of the protease MALT1 and cleavage of one of its targets - the ribonuclease Regnase-1- in lymphocytes and lymphoma cell lines

Persistent infection induces an adaptive immune response that is mediated by T and B lymphocytes. Upon triggering with an antigen, these cells become activated and turn into fast expanding cells able to efficiently defend the host. Lymphocyte activation is controlled by a complex composed of CARMA1, BCL10 and MALT1 which regulates the NF-KB signaling pathway upon antigen triggering. Abnormally high expression or activity of either one of these three proteins can favor the development of lymphomas, while genetic defects in the pathway are associated with immunodeficiency. MALT1 was identified as a paracaspase sharing homology with other cysteine proteases, namely caspases and metacaspases. In order to be active, caspases need to dimerize. Based on their sequence similarity with MALT1, we hypothesized that dimerization might also be a mechanism of activation employed by MALT1. To address this assumption, we performed a bioinformatics modelling based on the crystal structures of several caspases. Our model suggested that the MALT1 caspase-like domain can indeed form dimers. This finding was later confirmed by several published crystal structures of MALT1. In the dimer interface of our model, we noticed the presence of charged amino acids that could potentially form salt bridges and thereby hold both monomers together. Mutation of one of these residues, E549, into alanine completely blocked the catalytic activity of MALT1. Additionally, we provided evidence for a role of E549 in promoting the MALTl-dependent growth of cells derived from diffuse large B cell lymphoma (DLBCL) of the aggressive B cell-like type (ABC). To our initial surprise, the E549A mutation showed only a partial defect in dimerization, indicating that additional residues are essential to form a stable dimer. The MALT1 crystal structures revealed a key function for E549 in stabilizing the catalytic site of the protease via its interaction with an arginine which is located next to the catalytic active cysteine. In an additional study, we discovered that MALT1 monoubiquitination is required for the catalytic activity of the protease. Interestingly, we found that the MALT1 dimer interface mutant E549A could not be monoubiquitinated. Based on these findings, we suggest that correct formation of the dimer interface is a prerequisite for monoubiquitination. In a second project, we discovered a novel target of the protease MALT1, the ribonuclease Regnase¬la It was described that the RNase activity of Regnase-1 negatively regulates immune responses. We could show that in ABC DLBCL cell lines, Regnase-1 is not only cleaved by MALT1 but also phosphorylated, at least in part, by the inhibitor of KB kinase (IKK). Both regulations appear to restrain the RNase function of Regnase-1 and thereby allow the production of pro-survival proteins. In conclusion, our studies further highlight and explain the importance of the catalytic activity of MALT1 for the activation of lymphocytes and provide additional knowledge for the development of specific drugs targeting the catalytic activity of MALT1 for immunomodulation and treatment of lymphomas.  SUMMARY IN FRENCH PhD Thesis Katrin Cabalzar 2 SUMMARY IN FRENCH Une infection persistante induit une réponse immunitaire adaptative par l'intermédiaire des lymphocytes T et B. Quand elles reconnaissent l'antigène, ces cellules sont activées et se multiplient très rapidement pour défendre efficacement l'hôte. L'activation des lymphocytes est transmise par un complexe composé de trois protéines, CARMA1, BCL10 et MALT1, qui régule la voie de signalisation NF-KB lorsque l'antigène est reconnu. L'expression ou l'activité anormalement élevée de l'une de ces trois protéines peut favoriser le développement de lymphomes, tandis que des défauts génétiques de cette voie de signalisation sont associés à l'immunodéficience. MALT1 a été identifiée comme étant une paracaspase qui partage des séquences homologues avec d'autres protéases à cystéine, comme les caspases et les métacaspases. Pour être actives, les caspases ont besoin de dimériser. Etant donné leur similarité de séquence avec MALT1, nous avons supposé que la dimérisation pouvait aussi être un mécanisme d'activation utilisé par MALT1. Pour vérifier cette hypothèse, nous avons conçu un modèle bioinformatique à partir des structures cristallographiques de plusieurs caspases. Et notre modèle a suggéré que le domaine catalytique de MALT1 était effectivement capable de former des dimères. Cette découverte a été confirmée plus tard par des publications qui montrent des structures cristallographiques dimériques de MALT1. Dans l'interface du dimère de notre modèle, nous avons remarqué la présence d'acides aminés chargés qui pouvaient former des liaisons ioniques et ainsi réunir les deux monomères. La mutation de l'un de ces résidus, E549, pour une alanine, a complètement inhibé l'activité catalytique de MALT1. De plus, nous avons mis en évidence un rôle d'E549 dans la croissance dépendante de MALT1, des cellules dérivées de lymphomes B diffus à grandes cellules (DLBCL) de sous-type cellules B actives (ABC). Dans un premier temps nous avons été surpris de constater que cette mutation révélait seulement un défaut partiel de dimérisation, ce qui indique que des acides aminés supplémentaires sont indispensables pour former un dimère stable. Les structures cristallographiques de MALT1 ont révélé un rôle primordial d'E549 dans la stabilisation du site catalytique de la protéase via son interaction avec une arginine qui se trouve à côté de la cystéine du site actif. Dans une autre étude, nous avons découvert que la monoubiquitination de MALT1 est requise pour l'activité catalytique de la protéase. A remarquer que nous avons trouvé que le mutant E549A de l'interface dimère de MALT1 n'a pas pu être monoubiquitiné. Sur la base de ces résultats, nous suggérons que la formation correcte de l'interface du dimère est une condition préalable pour la monoubiquitination. Dans un second projet, nous avons découvert une nouvelle cible de la protéase MALT1, la ribonucléase Regnase-1. Il a été décrit que l'activité RNase de Regnase-1 régulait négativement les réponses immunitaires. Nous avons pu montrer que dans les lignées cellulaires ABC DLBCL, la Regnase-1 n'était pas seulement clivée par MALT1 mais également phosphorylée, au moins en partie, par la kinase de l'inhibiteur de KB (IKK). Les deux régulations semblent supprimer la fonction RNase de Regnase-1 et permettre ainsi la stabilisation de certains ARN messagers et la production de protéines favorisant la survie. En conclusion, nos études mettent en évidence le rôle-clé de la dimérisation de MALT1 et expliquent l'importance de l'activité catalytique de MALT1 pour l'activation des lymphocytes. Ainsi, nos résultats apportent des connaissances supplémentaires pour le développement de médicaments spécifiques ciblant l'activité catalytique de MALT1, qui pourraient être utiles pour modifier les réponses immunitaires et traiter des lymphomes.

Iowa Smart Planning Task Force Report

In the 2010 legislative session, the General Assembly passed and the Governor signed into law Senate File 2389 (SF 2389), which provided guidance for Smart Planning in Iowa and established the Iowa Smart Planning Task Force. This Task Force was charged with recommending policies and strategies for creating a stronger planning culture in Iowa, producing more resilient and sustainable communities. The Task Force, along with its two committees and four workgroups, met throughout the summer and fall of 2010 to identify and review best practices, consult local and national experts, and craft recommendations in the best interest of Iowans. A public input process was also implemented, resulting in improved recommendations.

Supplement Information to the August 2008 Economic and Workforce Development Task Force Report, August 2008

The 2008 disasters devastated businesses, farms, homes, schools, non-profit institutions, entire communities, and people’s lives across the state of Iowa. The Rebuild Iowa Advisory Commission (RIAC) is charged by the Governor to guide the state’s recovery and reconstruction process. The Economic and Workforce Development Task Force is respectfully submitting this report to be included and considered in the deliberations of the RIAC. While economic and workforce development are two issues that are inextricably linked and critical to Iowa’s rebuilding strategies, each also requires extraordinary attention in determining what needs to be considered in the very immediate and longer-term recovery. Supplement Information to the August 2008 Economic and Workforce Development Task Force Report

Supplemental Information to the August 2008 Long Term Recovery Planning Task Force Report, August 2008

Eighty-five of 99 Iowa counties were declared Presidential Disaster Areas for Public Assistance and/orIndividual Assistance as a result of the tornadoes, storms, and floods over the incident period May 25 through August 13, 2008. Response dominated the state’s attention for weeks, with a transition to recovery as the local situations warranted. The widespread damage and severity of the impact on Iowans and their communities required a statewide effort to continue moving forward despite being surrounded by adversity. By all accounts, it will require years for the state to recover from these disasters. With an eye toward the future, recovery is underway across Iowa. As part of the Rebuild Iowa efforts, the Long Term Recovery Planning Task Force was charged with responsibilities somewhat different from other topical Task Force assignments. Rather than assess damage and report on how the state might address immediate needs, the Long Term Recovery Planning Task Force is directed to discuss and discern the best approach to the lengthy recovery process. Certainly, the Governor and Lieutenant Governor expect the task to be difficult; when planning around so many critical issues and overwhelming needs, it is challenging to think to the future, rather than to rise to the current day’s needs.

Conduction mechanisms and charge storage in Si-nanocrystals metal-oxide-semiconductor memory devices studied with conducting atomic force microscopy

In this work, we demonstrate that conductive atomic force microscopy (C-AFM) is a very powerful tool to investigate, at the nanoscale, metal-oxide-semiconductor structures with silicon nanocrystals (Si-nc) embedded in the gate oxide as memory devices. The high lateral resolution of this technique allows us to study extremely small areas ( ~ 300nm2) and, therefore, the electrical properties of a reduced number of Si-nc. C-AFM experiments have demonstrated that Si-nc enhance the gate oxide electrical conduction due to trap-assisted tunneling. On the other hand, Si-nc can act as trapping centers. The amount of charge stored in Si-nc has been estimated through the change induced in the barrier height measured from the I-V characteristics. The results show that only ~ 20% of the Si-nc are charged, demonstrating that the electrical behavior at the nanoscale is consistent with the macroscopic characterization.

Carboxyl terminus structural requirements for glycosyl-phosphatidylinositol anchor addition to cell surface proteins.

Glycosyl phosphatidylinositol (GPI)-anchored proteins contain in their COOH-terminal region a peptide segment that is thought to direct glycolipid addition. This signal has been shown to require a pair of small amino acids positioned 10-12 residues upstream of an hydrophobic C-terminal domain. We analysed the contribution of the region separating the anchor acceptor site and the C-terminal hydrophobic segment by introducing amino acid deletions and substitutions in the spacer element of the GPI-anchored Thy-1 glycoprotein. Deletions of 7 amino acids in this region, as well as the introduction of 2 charged residues, prevented the glycolipid addition to Thy-1, suggesting that the length and the primary sequence of the spacer domain are important determinants in the signal directing GPI anchor transfer onto a newly synthesized polypeptide. Furthermore, we tested these rules by creating a truncated form of the normally transmembranous Herpes simplex virus I glycoprotein D (gDI) and demonstrating that when its C-terminal region displays all the features of a GPI-anchored protein, it is able to direct glycolipid addition onto another cell surface molecule.

Density-functional calculations of magnetoplasmons in quantum rings

We have studied the structure and dipole charge-density response of nanorings as a function of the magnetic field using local-spin-density-functional theory. Two small rings consisting of 12 and 22 electrons confined by a positively charged background are used to represent the cases of narrow and wide rings. The results are qualitatively compared with experimental data existing on microrings and on antidots. A smaller ring containing five electrons is also analyzed to allow for a closer comparison with a recent experiment on a two-electron quantum ring.