Extractants to assess zinc phytoavailability in mineral fertilizer and industrial by-products

Efficient analytical methods for the quantification of plant-available Zn contained in mineral fertilizers and industrial by-products are fundamental for the control and marketing of these inputs. In this sense, there are some doubts on the part of the scientific community as well as of the fertilizer production sector, whether the extractor requested by the government (Normative Instruction No. 28, called 2nd extractor), which is citric acid 2 % (2 % CA) (Brasil, 2007b), is effective in predicting the plant availability of Zn via mineral fertilizers and about the agronomic significance of the required minimal solubility of 60 % compared to the total content (HCl) (Brasil, 2007a). The purpose of this study was to evaluate the alternative extractors DTPA, EDTA, neutral ammonium citrate (NAC), buffer solution pH 6.0, 10 % HCl, 10 % sulfuric acid, 1 % acetic acid, water, and hot water to quantify the contents of Zn available for maize and compare them with indices of agronomic efficiency of fertilizers and industrial by-products when applied to dystrophic Clayey Red Latosol and Dystrophic Alic Red Yellow Latosol with medium texture. The rate of Zn applied to the soil was 5 mg kg-1, using the sources zinc sulfate, commercial granular zinc, ash and galvanic sludge, ash and two brass slags. Most Zn was extracted from the sources by DTPA, 10 % HCl, NAC, 1% acetic acid, and 10 % sulfuric acid. Recovery by the extractors 2 % CA, EDTA, water, and hot water was low. The agronomic efficiency index was found to be high when using galvanic sludge (238 %) and commercial granular zinc (142 %) and lower with brass slag I and II (67 and 27 %, respectively). The sources galvanizing ash and brass ash showed solubility lower than 60 % in 2 % CA, despite agronomic efficiency indices of 78 and 125 %, respectively. The low agronomic efficiency index of industrial by-products such as brass slag I and galvanizing ash can be compensated by higher doses, provided there is no restriction, as well as for all other sources, in terms of contaminant levels of arsenic, cadmium, chromium, lead, and mercury as required by law (Normative Instruction No 27/2006). The implementation of 2nd extractor 2 % CA and the requirement of minimum solubility for industrial by-products could restrict the use of alternative sources as potential Zn sources for plants.

Transparent conducting thin films by the co-sputtering of ZnO-ITO targets

Transparent and conductive Zn-In-Sn-O (ZITO) amorphous thin films have been deposited at room temperature by the rf magnetron co-sputtering of ITO and ZnO targets. Co-sputtering gives the possibility to deposit multicomponent oxide thin films with different compositions by varying the power to one of the targets. In order to make ZITO films with different Zn content, a constant rf power of 50 W was used for the ITO target, where as the rf power to ZnO target was varied from 25 W to 150 W. The as deposited films showed an increase in Zn content ratio from 17 to 67 % as the power to ZnO target was increased from 25 to 150 W. The structural, electrical and optical properties of the as deposited films are reported. The films showed an average transmittance over 80% in the visible wavelength range. The electrical resistivity and optical band gap of the ZITO films were found to depend on the Zn content in the film. The ZITO films deposited at room temperature with lower Zn content ratios showed better optical transmission and electrical properties compared to ITO film.

Electrical and optical properties of Zn-In-Sn-O transparent conducting thin films

Indium tin oxide (ITO) is one of the widely used transparent conductive oxides (TCO) for application as transparent electrode in thin film silicon solar cells or thin film transistors owing to its low resistivity and high transparency. Nevertheless, indium is a scarce and expensive element and ITO films require high deposition temperature to achieve good electrical and optical properties. On the other hand, although not competing as ITO, doped Zinc Oxide (ZnO) is a promising and cheaper alternative. Therefore, our strategy has been to deposit ITO and ZnO multicomponent thin films at room temperature by radiofrequency (RF) magnetron co-sputtering in order to achieve TCOs with reduced indium content. Thin films of the quaternary system Zn-In-Sn-O (ZITO) with improved electrical and optical properties have been achieved. The samples were deposited by applying different RF powers to ZnO target while keeping a constant RF power to ITO target. This led to ZITO films with zinc content ratio varying between 0 and 67%. The optical, electrical and morphological properties have been thoroughly studied. The film composition was analysed by X-ray Photoelectron Spectroscopy. The films with 17% zinc content ratio showed the lowest resistivity (6.6 × 10 - 4 Ω cm) and the highest transmittance (above 80% in the visible range). Though X-ray Diffraction studies showed amorphous nature for the films, using High Resolution Transmission Electron Microscopy we found that the microstructure of the films consisted of nanometric crystals embedded in a compact amorphous matrix. The effect of post deposition annealing on the films in both reducing and oxidizing atmospheres were studied. The changes were found to strongly depend on the zinc content ratio in the films.

Uniformity Study of Amorphous and Microcrystalline Silicon Thin Films Deposited on 10cmx10cm Glass Substrate using Hot Wire CVD Technique

The scaling up of the Hot Wire Chemical Vapor Deposition (HW-CVD) technique to large deposition area can be done using a catalytic net of equal spaced parallel filaments. The large area deposition limit is defined as the limit whenever a further increment of the catalytic net area does not affect the properties of the deposited film. This is the case when a dense catalytic net is spread on a surface considerably larger than that of the film substrate. To study this limit, a system able to hold a net of twelve wires covering a surface of about 20 cm x 20 cm was used to deposit amorphous (a-Si:H) and microcrystalline (μc-Si:H) silicon over a substrate of 10 cm x 10 cm placed at a filament-substrate distance ranging from 1 to 2 cm. The uniformity of the film thickness d and optical constants, n(x, λ) and α(x,¯hω), was studied via transmission measurements. The thin film uniformity as a function of the filament-substrate distance was studied. The experimental thickness profile was compared with the theoretical result obtained solving the diffusion equations. The optimization of the filament-substrate distance allowed obtaining films with inhomogeneities lower than ±2.5% and deposition rates higher than 1 nm/s and 4.5 nm/s for (μc-Si:H) and (a-Si:H), respectively.

The role of fungi in the precipitation of calcite : relationships between fungal filaments, nanofibres, and needle fibre calcite

RésuméLes champignons sont impliqués dans les cycles biogéochimiques de différentes manières. En particulier, ils sont reconnus en tant qu'acteurs clés dans la dégradation de la matière organique, comme fournisseurs d'éléments nutritifs via l'altération des minéraux mais aussi comme grands producteurs d'acide oxalique et de complexes oxalo-métalliques. Toutefois, peu de choses sont connues quant à leur contribution à la genèse d'autres types de minéraux, tel que le carbonate de calcium (CaCO3). Le CaCO3 est un minéral ubiquiste dans de nombreux écosystèmes et il joue un rôle essentiel dans les cycles biogéochimiques du carbone (C) et du calcium (Ca). Le CaCO3 peut être d'origine physico-chimique ou biogénique et de nombreux organismes sont connus pour contrôler ou induire sa biominéralisation. Les champignons ont souvent été soupçonnés d'être impliqué dans ce processus, cependant il existe très peu d'informations pour étayer cette hypothèse.Cette thèse a eu pour but l'étude de cet aspect négligé de l'impact des champignons dans les cycles biogéochimiques, par l'exploration de leur implication potentielle dans la formation d'un type particulier de CaCO3 secondaires observés dans les sols et dans les grottes des environnements calcaires. Dans les grottes, ces dépôts sont appelés moonmilk, alors que dans les sols on les appelle calcite en aiguilles. Cependant ces deux descriptions correspondent en fait au même assemblage microscopique de deux habitus particulier de la calcite: la calcite en aiguilles (au sens strict du terme cette fois-ci) et les nanofibres. Ces deux éléments sont des habitus aciculaires de la calcite, mais présentent des dimensions différentes. Leur origine, physico-chimique ou biologique, est l'objet de débats intenses depuis plusieurs années déjà.L'observation d'échantillons environnementaux avec des techniques de microscopie (microscopie électronique et micromorphologie), ainsi que de la microanalyse EDX, ont démontré plusieurs relations intéressantes entre la calcite en aiguilles, les nanofibres et des éléments organiques. Premièrement, il est montré que les nanofibres peuvent être organiques ou minérales. Deuxièmement, la calcite en aiguilles et les nanofibres présentent de fortes analogies avec des structures hyphales, ce qui permet de confirmer l'hypothèse de leur origine fongique. En outre, des expériences en laboratoire ont confirmé l'origine fongique des nanofibres, par des digestions enzymatiques d'hyphes fongiques. En effet, des structures à base de nanofibres, similaires à celles observées dans des échantillons naturels, ont pu être produites par cette approche. Finalement, des enrichissements en calcium ont été mesurés dans les parois des hyphes et dans des inclusions intrahyphales provenant d'échantillons naturels de rhizomorphes. Ces résultats suggèrent une implication de la séquestration de calcium dans la formation de la calcite en aiguilles et/ou des nanofibres.Plusieurs aspects restent à élucider, en particulier la compréhension des processus physiologiques impliqués dans la nucléation de calcite dans les hyphes fongiques. Cependant, les résultats obtenus dans cette thèse ont permis de confirmer l'implication des champignons dans la formation de la calcite en aiguilles et des nanofibres. Ces découvertes sont d'une grande importance dans les cycles biogéochimiques puisqu'ils apportent de nouveaux éléments dans le cycle couplé C-Ca. Classiquement, les champignons sont considérés comme étant impliqués principalement dans la minéralisation de la matière organique et dans l'altération minérale. Cette étude démontre que les champignons doivent aussi être pris en compte en tant qu'agents majeurs de la genèse de minéraux, en particulier de CaCO3. Ceci représente une toute nouvelle perspective en géomycologie quant à la participation des champignons au cycle biologique du C. En effet, la présence de ces précipitations de CaCO3 secondaires représente un court-circuit dans le cycle biologique du C puisque du C inorganique du sol se retrouve piégé dans de la calcite plutôt que d'être retourné dans l'atmosphère.AbstractFungi are known to be involved in biogeochemical cycles in numerous ways. In particular, they are recognized as key players in organic matter recycling, as nutrient suppliers via mineral weathering, as well as large producers of oxalic acid and metal-oxalate. However, little is known about their contribution to the genesis of other types of minerals such as calcium carbonate (CaCO3). Yet, CaC03 are ubiquitous minerals in many ecosystems and play an essential role in the biogeochemical cycles of both carbon (C) and calcium (Ca). CaC03 may be physicochemical or biogenic in origin and numerous organisms have been recognized to control or induce calcite biomineralization. While fungi have often been suspected to be involved in this process, only scarce information support this hypothesis.This Ph.D. thesis aims at investigating this disregarded aspect of fungal impact on biogeochemical cycles by exploring their possible implication in the formation of a particular type of secondary CaC03 deposit ubiquitously observed in soils and caves from calcareous environments. In caves, these deposits are known as moonmilk, whereas in soils, they are known as Needle Fibre Calcite (NFC - sensu lato). However, they both correspond to the same microscopic assemblage of two distinct and unusual habits of calcite: NFC {sensu stricto) and nanofibres. Both features are acicular habits of calcite displaying different dimensions. Whether these habits are physicochemical or biogenic in origin has been under discussion for a long time.Observations of natural samples using microscopic techniques (electron microscopy and micromorphology) and EDX microanalyses have demonstrated several interesting relationships between NFC, nanofibres, and organic features. First, it has shown that nanofibres can be either organic or minera! in nature. Second, both nanofibres and NFC display strong structural analogies with fungal hyphal features, supporting their fungal origin. Furthermore, laboratory experiments have confirmed the fungal origin of nanofibres through an enzymatic digestion of fungal hyphae. Indeed, structures made of nanofibres with similar features as those observed in natural samples have been produced. Finally, calcium enrichments have been measured in both cell walls and intrahyphal inclusions of hyphae from rhizomorphs sampled in the natural environment. These results point out an involvement of calcium sequestration in nanofibres and/or NFC genesis.Several aspects need further investigation, in particular the understanding of the physiological processes involved in hyphal calcite nucleation. However, the results obtained during this study have allowed the confirmation of the implication of fungi in the formation of both NFC and nanofibres. These findings are of great importance regarding global biogeochemical cycles as they bring new insights into the coupled C and Ca cycles. Conventionally, fungi are considered to be involved in organic matter mineralization and mineral weathering. In this study, we demonstrate that they must also be considered as major agents in mineral genesis, in particular CaC03. This is a completely new perspective in geomycology regarding the role of fungi in the short-term (or biological) C cycle. Indeed, the presence of these secondary CaC03 precipitations represents a bypass in the short- term carbon cycle, as soil inorganic C is not readily returned to the atmosphere.

Influence of digestion methods on the recovery of Iron, Zinc, Nickel, Chromium, Cadmium and Lead contents in 11 organic residues

There are currently many devices and techniques to quantify trace elements (TEs) in various matrices, but their efficacy is dependent on the digestion methods (DMs) employed in the opening of such matrices which, although "organic", present inorganic components which are difficult to solubilize. This study was carried out to evaluate the recovery of Fe, Zn, Cr, Ni, Cd and Pb contents in samples of composts and cattle, horse, chicken, quail, and swine manures, as well as in sewage sludges and peat. The DMs employed were acid digestion in microwaves with HNO3 (EPA 3051A); nitric-perchloric digestion with HNO3 + HClO4 in a digestion block (NP); dry ashing in a muffle furnace and solubilization of residual ash in nitric acid (MDA); digestion by using aqua regia solution (HCl:HNO3) in the digestion block (AR); and acid digestion with HCl and HNO3 + H2O2 (EPA 3050). The dry ashing method led to the greatest recovery of Cd in organic residues, but the EPA 3050 protocol can be an alternative method for the same purpose. The dry ashing should not be employed to determine the concentration of Cr, Fe, Ni, Pb and Zn in the residues. Higher Cr and Fe contents are recovered when NP and EPA 3050 are employed in the opening of organic matrices. For most of the residues analyzed, AR is the most effective method for recovering Ni. Microwave-assisted digestion methods (EPA3051 and 3050) led to the highest recovery of Pb. The choice of the DM that provides maximum recovery of Zn depends on the organic residue and trace element analyzed.

Rice straw incorporated just before soil flooding increases acetic acid formation and decreases available nitrogen

Incorporation of rice straw into the soil just before flooding for water-seeded rice can immobilize mineral nitrogen (N) and lead to the production of acetic acid harmful to the rice seedlings, which negatively affects grain yield. This study aimed to evaluate the formation of organic acids and variation in pH and to quantify the mineral N concentration in the soil as a function of different times of incorporation of rice straw or of ashes from burning the straw before flooding. The experiment was carried out in a greenhouse using an Inceptisol (Typic Haplaquept) soil. The treatments were as follows: control (no straw or ash); incorporation of ashes from previous straw burning; rice straw incorporated to drained soil 60 days before flooding; straw incorporated 30 days before flooding; straw incorporated 15 days before flooding and straw incorporated on the day of flooding. Experimental units were plastic buckets with 6.0 kg of soil. The buckets remained flooded throughout the trial period without rice plants. Soil samples were collected every seven days, beginning one day before flooding until the 13th week of flooding for determination of mineral N- ammonium (NH4+) and nitrate (NO3-). Soil solution pH and concentration of organic acids (acetic, propionic and butyric) were determined. All NO3- there was before flooding was lost in approximately two weeks of flooding, in all treatments. There was sigmoidal behavior for NH4+ formation in all treatments, i.e., ammonium ion concentration began to rise shortly after soil flooding, slightly decreased and then went up again. On the 91st day of flooding, the NH4+ concentrations in soil was 56 mg kg-1 in the control treatment, 72 mg kg-1 for the 60-day treatment, 73 mg kg-1 for the 30-day treatment and 53 mg kg-1 for the ash incorporation treatment. These ammonium concentrations correspond to 84, 108, 110 and 80 kg ha-1 of N-NH4+, respectively. When the straw was incorporated on the day of flooding or 15 days before, the concentration of N-NH4+ in the soil was 28 and 54 mg kg-1, equivalent to an accumulation of 42 and 81 kg ha-1 of N-NH4+, respectively. There was formation of acetic acid in which toxic concentrations were reached (7.2 mmol L-1) on the 15th day of flooding only for the treatment with straw incorporated on the day of flooding. The pH of the soil solution of all the treatments increased after flooding and this increase was faster in the treatments with incorporation of straw, followed by the ash treatment and then the control. After 60 days of flooding, however, the pH values were around 6.5 for all treatments, except for the control, which reached a pH of 6.3. Rice straw should be incorporated into the soil at least 30 days before flooding; otherwise, it may immobilize part of the mineral N and produce acetic acid in concentrations toxic to rice seedlings.

Evolution of a Ladinian (Middle Triassic) intraplatform basin : stratigraphy, microfacies and palaeoecology of the Meride Limestone (Monte San Giorgio, Canton Ticino, Southern Switzerland)

The Monte San Giorgio (Southern Alps, Ticino, Switzerland) is the most important locality in the world for vertebrates dating back to the Middle Triassic. For this reason it was registered in 2003 as a UNESCO World Heritage Site. One of the objectives of this doctoral thesis was to fill some of the cognitive gaps regarding the Ladinian succession, including in particular the San Giorgio Dolomite and the Meride Limestone. In order to achieve this, the entire succession, more than 600 metres thick, was measured and sampled. Biostratigraphic research based on new finds of fossil invertebrates and microfossils and on the palynological analysis of the entire section was integrated with single-zircon U-Pb dating of volcanic ash layers intercalated in the carbonate succession. This enabled a redefinition of the bio-chronostratigraphic and geochronologic framework of the succession, which encompasses a significantly shorter time interval than previously held. The Ladinian section extends from the E. curionii Ammonoid Zone (Early Fassanian) to the P. archelaus Ammonoid Zone (Early Longobardian). The age of the classic fossiliferous levels of the Meride Limestone, rich in organic matter and containing vertebrate fossils which are known all over the world, was defined in both biostratigraphic and geochronologic terms. The presumed stratigraphie significance of the pachypleurosaurid reptiles found in such levels is called into question by new finds. These fossiliferous horizons were found to correspond to the main volcanoclastic intervals of the Buchenstein Formation (Middle and Upper Pietra Verde). Thus, a correlation with the Bagolino Section (Italy) containing the GSSP for the base of the Ladinian was proposed. Bulk sedimentation rates in the studied succession average 200 m/Myr and therefore prove to be 20 times higher than those of the South-Alpine pelagic basins. These values express high carbonate productivity from the surrounding platforms on one hand, and on the other a marked subsidence of the basin. Only in the intervals consisting of laminated limestones did the sedimentation rates drop to average values of around 30 m/Myr. The distribution of organic and inorganic facies appears to be the consequence of relative variations in sea-level. The laminated and organic-matter- rich intervals of the Meride Limestone are linked to a relative sea-level drop which favoured dysoxic to anoxic bottom-water conditions, coupled with an increase in runoff, perhaps due to recurrent explosive volcanic activity. The transient development under dysoxic conditions of monospecific benthic meio-/macrofaunas was documented. Organic matter suggests a predominant origin due to benthic bacterial activity, as can be witnessed in alveolar structures typical of exopolymeric substances secreted by bacteria within microbial mats. A microbial contribution to the carbonate (peloidal) precipitation was documented. The protective effect exerted by these microbial mats is also indicated as the main taphonomic factor contributing to the excellent preservation of vertebrate fossils. A radiolarian assemblage discovered in the lower part of the section (earliest Ladinian, E. curionii Zone) suggests the transient existence of open-marine but not deep-water connections with the tethyan pelagic basins. It shows marked similarities to the faunas typical of the late Anisian, suggesting therefore a low resolution power provided by radiolarian biostratigraphy in recognizing the Anisian/Ladinian boundary. The present thesis describes a new species of conifer (Elatocladus cassinae), a new species of insect (Dasyleptus triassicus) and seven new species of radiolarians (Eptingium danieli, Eptingium neriae, Parentactinosphaera eoladinica, Sepsagon ticinensis, Sepsagon? valporinae, Novamuria wirzi and Pessagnollum? hexaspinosum). In addition, following revision of the type material of already existent taxa, four new genera of radiolarians are introduced: Bernoulliella, Eohexastylus, Ticinosphaera and Lahmosphaera.

Properties of debris flow deposits and source materials compared: implications for debris flow characterization

In the Alps, debris flow deposits generally contain < 5% clay-size particles, and the role of the surface-charged < 2 mu m particles is often neglected, although these particles may have a significant impact on the rheological properties of the interstitial fluid. The objective of this study was to compare debris flow deposits and parent materials from two neighbouring catchments of the Swiss Alps, with special emphasis on the colloidal constituents. The catchments are small in area (4 km(2)), 2.5 km long, similar in morphology, but different in geology. The average slopes are 35-40%. The catchments were monitored for debris flow events and mapped for surface aspect and erosion activity. Debris flow deposits and parent materials were sampled, the clay and silt fractions extracted and the bulk density, < 2 mm fraction bulk density, particle size distribution, chemical composition, cation exchange capacity (CEC) and mineralogy analysed. The results show that the deposits are similar to the parent screes in terms of chemical composition, but differ in terms of: (i) particle size distribution; and (ii) mineralogy, reactivity and density of the < 2 mm fraction. In this fraction, compared with the parent materials the deposits show dense materials enriched in coarse monocrystalline particles, of which the smallest and more reactive particles were leached. The results suggest that deposit samples should not be considered as representative of source or flow materials, particularly with respect to their physical properties.

Detection and Characterization of Preferential Flow Paths in the Downstream Area of a Hazardous Landfill

We have used surface-based electrical resistivity tomography to detect and characterize preferential hydraulic pathways in the immediate downstream area of an abandoned, hazardous landfill. The landfill occupies the void left by a former gravel pit and its base is close to the groundwater table and lacking an engineered barrier. As such, this site is remarkably typical of many small- to medium-sized waste deposits throughout the densely populated and heavily industrialized foreland on both sides of the Alpine arc. Outflows of pollutants lastingly contaminated local drinking water supplies and necessitated a partial remediation in the form of a synthetic cover barrier, which is meant to prevent meteoric water from percolating through the waste before reaching the groundwater table. Any future additional isolation of the landfill in the form of lateral barriers thus requires adequate knowledge of potential preferential hydraulic pathways for outflowing contaminants. Our results, inferred from a suite of tomographically inverted surfaced-based electrical resistivity profiles oriented roughly perpendicular to the local hydraulic gradient, indicate that potential contaminant outflows would predominantly occur along an unexploited lateral extension of the original gravel deposit. This finds its expression as a distinct and laterally continuous high-resistivity anomaly in the resistivity tomograms. This interpretation is ground-truthed through a litholog from a nearby well. Since the probed glacio-fluvial deposits are largely devoid of mineralogical clay, the geometry of hydraulic and electrical pathways across the pore space of a given lithological unit can be assumed to be identical, which allows for an order-of-magnitude estimation of the overall permeability structure. These estimates indicate that the permeability of the imaged extension of the gravel body is at least two to three orders-of-magnitude higher than that of its finer-grained embedding matrix. This corroborates the preeminent role of the high-resistivity anomaly as a potential preferential flow path.

A Mechanistic Explanation of The Physical Properties of Undisturbed Loess, February, 1968

The relation between the properties and the water content of an undisturbed loess were investigated to provide insight into the mechanical behavior of the natural soil. Hand-carved samples from a single deposit, at their natural water contents, and at water contents modified in the laboratory to provide a range from 870 to 3270, were subjected to unconsolidated-undrained triaxial compression tests, consolidation tests, and initial negative pore water pressure tests. In addition, the clay-size fraction was separated from the remainder of the loess for a separate series of tests to establish its properties. The natural water content of the deposit in the field was measured at regular intervals for one year to provide an example of the range in properties that would be encountered. at this site. The test results are presented and their interpretation leads to conclusions regarding the volumetric relations that exist as the water content varies. The significance of the water content in relation to the properties of the natural soil is explored and the concept of a critical water content for loess is introduced.

Blood profile during and after hemoglobin substitute administration.

The in vivo effects of Diaspirin Crosslinked Hemoglobin (DCLHb, Baxter Healthcare Corp.) on hematology and biochemistry are unknown. This study includes 6 calves (71.2+/-1.3 kg). In each animal a total of 2 litres of blood was exchanged for the same amount of hydroxylethyl starch (Haes, Fresenius) (n=3) or DCLHb (n=3), which is equivalent to 28cc/kg of blood substitute, over a period of 5 hours. The animals were allowed to survive 7 days. Blood samples were taken hourly during the perfusion protocol, at postoperative day (POD) 1, 2 and 7. ANOVA test was used for repeated measurements. Blood cell profiles were similar in both groups. Peak methemoglobinemia was 4.2% in the DCLHb group. Osmolarity was significantly higher in the DCLHb group with the greatest difference at POD 1 and 2. Postmortem analysis of the major organs did not show any sign of hemoglobin deposit in the DCLHb group. In the given setup DCLHb can be administered in a large quantity with good hematological tolerance and without any deposits in major organs. A prolonged plasma expander effect was observed.

Insight in protein S deficiency from mouse models

Protein S (ProS) is an important negative regulator of blood coagulation. Its physiological importance is evident in purpura fulminans and other life-threatening thrombotic disorders typical of ProS deficient patients. Our previous characterization of ProS deficiency in mouse models has shown similarities with the human phenotypes: heterozygous ProS-deficient mice (Pros+/-) had increased thrombotic risk whereas homozygous deficiency in ProS (Pros-/-) was incompatible with life (Blood 2009; 114:2307-2314). In tissues, ProS exerts cellular functions by binding to and activating tyrosine kinase receptors of the Tyro3 family (TAM) on the cell surface.To extend the analysis of coagulation defects beyond the Pros-/- phenotype and add new insights into the sites of synthesis ProS and its action, we generated mice with inactivated ProS in hepatocytes (Proslox/loxAlbCre+) as well as in endothelial and hematopoietic cells (Proslox/loxTie2Cre+). Both models resulted in significant reduction of circulating ProS levels and in a remarkable increased thrombotic risk in vivo. In a model of tissue factor (TF)-induced venous thromboembolism (VTE), only 17% of Proslox/loxAlbCre+ mice (n=12) and only 13% of Proslox/loxTie2Cre+ mice (n=14) survived, compared with 86% of Proslox/lox mice (n=14; P<0.001).To mimic a severe acquired ProS deficiency, ProS gene was inactivated at the adult stage using the polyI:C-inducible Mx1-Cre system (Proslox/loxMx1Cre+). Ten days after polyI:C treatment, Proslox/loxMx1Cre+ mice developed disseminated intravascular coagulation with extensive lung and liver thrombosis.It is worth noting that no skin lesions compatible with purpura fulminans were observed in any of the above-described models of partial ProS deficiency. In order to shed light on the pathogenesis of purpura fulminans, we exposed the different ProS-deficient mice to warfarin (0.2 mg/day). We observed that Pros+/-, Proslox/loxAlbCre+ and Proslox/loxTie2Cre+ mice developed retiform purpura (characterized by erythematous and necrotic lesions of the genital region and extremities) and died after 3 to 5 days after the first warfarin administration.In human, ProS is also synthesized by megakaryocytes and hence stored at high concentrations in circulating platelets (pProS). The role of pProS has been investigated by generating megakaryocyte ProS-deficient model using the PF4 promoter as Cre driver (Proslox/loxPf4Cre+). In the TF-induced VTE model, Proslox/loxPf4Cre+ (n=15) mice showed a significant increased risk of thrombosis compared to Proslox/lox controls (n=14; survival rate 47% and 86%, respectively; P<0.05). Furthermore, preliminary results suggest survival to be associated with higher circulating ProS levels. In order to evaluate the potential role of pProS in thrombus formation, we investigated the thrombotic response to intravenous injection of collagen-epinephrine in vivo and platelet function in vitro. Both in vivo and in vitro experiments showed similar results between Proslox/loxPf4Cre+ and Proslox/lox, indicating that platelet reactivity was not influenced by the absence of pProS. These data suggest that pProS is delivered at the site of thrombosis to inhibit thrombin generation.We further investigated the ability of ProS to function as a ligand of TAM receptors, by using homozygous and heterozygous deficient mice for both the TAM ligands ProS and Gas6. Gas6-/-Pros-/- mice died in utero and showed comparable dramatic bleeding and thrombotic phenotype as described for Pros-/- embryos.In conclusion, like complete ProS deficiency, double deficiency in ProS and Gas6 was lethal, whereas partial ProS deficiency was not. Mice partially deficient in ProS displayed a prothrombotic phenotype, including those with only deficiency in pProS. Purpura fulminans did not occur spontaneously in mice with partial Pros deficiency but developed upon warfarin administration.Thus, the use of different mice models of ProS deficiency can be instrumental in the study of its highly variable thrombotic phenotype and in the investigation of additional roles of ProS in inflammation and autoimmunity through TAM signaling.

Further insights on dynamic morphological transitions in quasi-two-dimensional electrodeposition

Dynamic morphological transitions in thin-layer electrodeposits obtained from copper sulphate solutions have been studied. The chemical composition of the electrodeposits indicates that they appear as a consequence of the competition between copper and cuprous oxide formation. In addition, the Ohmic control of the process is verified at initial stages of the deposit growth. At higher deposit developments, gravity-induced convection currents play a role in the control of the whole process and affect the position of these transitions.

Morphological changes in convection-diffusion-limited deposition

The effect of hydrodynamic flow upon diffusion-limited deposition on a line is investigated using a Monte Carlo model. The growth process is governed by the convection and diffusion field. The convective diffusion field is simulated by the biased-random walker resulting from a superimposed drift that represents the convective flow. The development of distinct morphologies is found with varying direction and strength of drift. By introducing a horizontal drift parallel to the deposition plate, the diffusion-limited deposit changes into a single needle inclined to the plate. The width of the needle decreases with increasing strength of drift. The angle between the needle and the plate is about 45° at high flow rate. In the presence of an inclined drift to the plate, the convection-diffusion-limited deposit leads to the formation of a characteristic columnar morphology. In the limiting case where the convection dominates, the deposition process is equivalent to ballistic deposition onto an inclined surface.