Biochemical and metabolic profiles of Trichoderma strains isolated from common bean crops in the Brazilian Cerrado, and potential antagonism against Sclerotinia sclerotiorum

Some species of Trichoderma have successfully been used in the commercial biological control of fungal pathogens, e.g., Sclerotinia sclerotiorum, an economically important pathogen of common beans (Phaseolus vulgaris L.). The objectives of the present study were (1) to provide molecular characterization of Trichoderma strains isolated from the Brazilian Cerrado; (2) to assess the metabolic profile of each strain by means of Biolog FF Microplates; and (3) to evaluate the ability of each strain to antagonize S. sclerotiorum via the production of cell wall-degrading enzymes (CWDEs), volatile antibiotics, and dual-culture tests. Among 21 isolates, we identified 42.86 % as Trichoderma asperellum, 33.33 % as Trichoderma harzianum, 14.29 % as Trichoderma tomentosum, 4.76 % as Trichoderma koningiopsis, and 4.76 % as Trichoderma erinaceum. Trichoderma asperellum showed the highest CWDE activity. However, no species secreted a specific group of CWDEs. Trichoderma asperellum 364/01, T. asperellum 483/02, and T. asperellum 356/02 exhibited high and medium specific activities for key enzymes in the mycoparasitic process, but a low capacity for antagonism. We observed no significant correlation between CWDE and antagonism, or between metabolic profile and antagonism. The diversity of Trichoderma species, and in particular of T. harzianum, was clearly reflected in their metabolic profiles. Our findings indicate that the selection of Trichoderma candidates for biological control should be based primarily on the environmental fitness of competitive isolates and the target pathogen. (C) 2012 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Immunoglobulin G profile in hyperacute rejection after multivisceral xenotransplantation

Galvao FHF, Soler W, Pompeu E, Waisberg DR, Mello ES, Costa ACL, Teodoro W, Velosa AP, Capelozzi VL, Antonangelo L, Catanozi S, Martins A, Malbouisson LMS, Cruz RJ, Figueira ER, Filho JAR, Chaib E, D'Albuquerque LAC. Immunoglobulin G profile in hyperacute rejection after multivisceral xenotransplantation. Xenotransplantation 2012; 19: 298304. (c) 2012 John Wiley & Sons A/S. Abstract: Introduction: Xenotransplantation is a potential solution for the high mortality of patients on the waiting list for multivisceral transplantation; nevertheless, hyperacute rejection (HAR) hampers this practice and motivates innovative research. In this report, we describe a model of multivisceral xenotransplantation in which we observed immunoglobulin G (IgG) involvement in HAR. Methods: We recovered en bloc multivisceral grafts (distal esophagus, stomach, small intestine, colon, liver, pancreas, and kidneys) from rabbits (n = 20) and implanted them in the swine (n = 15) or rabbits (n = 5, control). Three hours after graft reperfusion, we collected samples from all graft organs for histological study and to assess IgG fixation by immunofluorescence. Histopathologic findings were graded according to previously described methods. Results: No histopathological features of rejection were seen in the rabbit allografts. In the swine-to-rabbit grafts, features of HAR were moderate in the liver and severe in esophagus, stomach, intestines, spleen, pancreas, and kidney. Xenograft vessels were the central target of HAR. The main lesions included edema, hemorrhage, thrombosis, myosites, fibrinoid degeneration, and necrosis. IgG deposition was intense on cell membranes, mainly in the vascular endothelium. Conclusions: Rabbit-to-swine multivisceral xenotransplants undergo moderate HAR in the liver and severe HAR in the other organs. Moderate HAR in the liver suggests a degree of resistance to the humoral immune response in this organ. Strong IgG fixation in cell membranes, including vascular endothelium, confirms HAR characterized by a primary humoral immune response. This model allows appraisal of HAR in multiple organs and investigation of the livers relative resistance to this immune response.

The implementation of the Medical Regulation Office and Mobile Emergency Attendance System and its impact on the gravity profile of non-traumatic afflictions treated in a University Hospital: a research study

Abstract Background The public health system of Brazil is structured by a network of increasing complexity, but the low resolution of emergency care at pre-hospital units and the lack of organization of patient flow overloaded the hospitals, mainly the ones of higher complexity. The knowledge of this phenomenon induced Ribeirão Preto to implement the Medical Regulation Office and the Mobile Emergency Attendance System. The objective of this study was to analyze the impact of these services on the gravity profile of non-traumatic afflictions in a University Hospital. Methods The study conducted a retrospective analysis of the medical records of 906 patients older than 13 years of age who entered the Emergency Care Unit of the Hospital of the University of São Paulo School of Medicine at Ribeirão Preto. All presented acute non-traumatic afflictions and were admitted to the Internal Medicine, Surgery or Neurology Departments during two study periods: May 1996 (prior to) and May 2001 (after the implementation of the Medical Regulation Office and Mobile Emergency Attendance System). Demographics and mortality risk levels calculated by Acute Physiology and Chronic Health Evaluation II (APACHE II) were determined. Results From 1996 to 2001, the mean age increased from 49 ± 0.9 to 52 ± 0.9 (P = 0.021), as did the percentage of co-morbidities, from 66.6 to 77.0 (P = 0.0001), the number of in-hospital complications from 260 to 284 (P = 0.0001), the mean calculated APACHE II mortality risk increased from 12.0 ± 0.5 to 14.8 ± 0.6 (P = 0.0008) and mortality rate from 6.1 to 12.2 (P = 0.002). The differences were more significant for patients admitted to the Internal Medicine Department. Conclusion The implementation of the Medical Regulation and Mobile Emergency Attendance System contributed to directing patients with higher gravity scores to the Emergency Care Unit, demonstrating the potential of these services for hierarchical structuring of pre-hospital networks and referrals.

MicroRNA expression profile in head and neck cancer: HOX-cluster embedded microRNA-196a and microRNA-10b dysregulation implicated in cell proliferation

Abstract Background Current evidence implicates aberrant microRNA expression patterns in human malignancies; measurement of microRNA expression may have diagnostic and prognostic applications. Roles for microRNAs in head and neck squamous cell carcinomas (HNSCC) are largely unknown. HNSCC, a smoking-related cancer, is one of the most common malignancies worldwide but reliable diagnostic and prognostic markers have not been discovered so far. Some studies have evaluated the potential use of microRNA as biomarkers with clinical application in HNSCC. Methods MicroRNA expression profile of oral squamous cell carcinoma samples was determined by means of DNA microarrays. We also performed gain-of-function assays for two differentially expressed microRNA using two squamous cell carcinoma cell lines and normal oral keratinocytes. The effect of the over-expression of these molecules was evaluated by means of global gene expression profiling and cell proliferation assessment. Results Altered microRNA expression was detected for a total of 72 microRNAs. Among these we found well studied molecules, such as the miR-17-92 cluster, comprising potent oncogenic microRNA, and miR-34, recently found to interact with p53. HOX-cluster embedded miR-196a/b and miR-10b were up- and down-regulated, respectively, in tumor samples. Since validated HOX gene targets for these microRNAs are not consistently deregulated in HNSCC, we performed gain-of-function experiments, in an attempt to outline their possible role. Our results suggest that both molecules interfere in cell proliferation through distinct processes, possibly targeting a small set of genes involved in cell cycle progression. Conclusions Functional data on miRNAs in HNSCC is still scarce. Our data corroborate current literature and brings new insights into the role of microRNAs in HNSCC. We also show that miR-196a and miR-10b, not previously associated with HNSCC, may play an oncogenic role in this disease through the deregulation of cell proliferation. The study of microRNA alterations in HNSCC is an essential step to the mechanistic understanding of tumor formation and could lead to the discovery of clinically relevant biomarkers.

Natural blood feeding and temperature shift modulate the global transcriptional profile of Rickettsia rickettsii infecting its tick vector

Rickettsia rickettsii is an obligate intracellular tick-borne bacterium that causes Rocky Mountain Spotted Fever (RMSF), the most lethal spotted fever rickettsiosis. When an infected starving tick begins blood feeding from a vertebrate host, R. rickettsii is exposed to a temperature elevation and to components in the blood meal. These two environmental stimuli have been previously associated with the reactivation of rickettsial virulence in ticks, but the factors responsible for this phenotype conversion have not been completely elucidated. Using customized oligonucleotide microarrays and high-throughput microfluidic qRT-PCR, we analyzed the effects of a 10 degrees C temperature elevation and of a blood meal on the transcriptional profile of R. rickettsii infecting the tick Amblyomma aureolatum. This is the first study of the transcriptome of a bacterium in the genus Rickettsia infecting a natural tick vector. Although both stimuli significantly increased bacterial load, blood feeding had a greater effect, modulating five-fold more genes than the temperature upshift. Certain components of the Type IV Secretion System (T4SS) were up-regulated by blood feeding. This suggests that this important bacterial transport system may be utilized to secrete effectors during the tick vector's blood meal. Blood feeding also up-regulated the expression of antioxidant enzymes, which might correspond to an attempt by R. rickettsii to protect itself against the deleterious effects of free radicals produced by fed ticks. The modulated genes identified in this study, including those encoding hypothetical proteins, require further functional analysis and may have potential as future targets for vaccine development.

Variability of the phenolic profile of banana from Gran Canaria during maturation

[EN] The identification and quantification of phenolic compounds in bananas from Gran Canaria were carried out to examine changes in their profile during maturation. The phenolic composition of extracts derived from bananas was determined by using HPLC. The total phenolic contents were measured with Folin–Cicolteau`s phenol reagent. To quantify the total antioxidant capacity of banana extracts, we chose two methods commonly used: the percentage of inhibition of radical 1,1–diphenyl-2-picrylhydrazyl (DPPH) and the ferric reducing antioxidant power (FRAP assay). The antioxidant potential on banana extracts was found to increase during maturation in correlation whit the total phenol content.

Bioremediation of PAHs - Limitations and soultions

Introduction 1.1 Occurrence of polycyclic aromatic hydrocarbons (PAH) in the environment Worldwide industrial and agricultural developments have released a large number of natural and synthetic hazardous compounds into the environment due to careless waste disposal, illegal waste dumping and accidental spills. As a result, there are numerous sites in the world that require cleanup of soils and groundwater. Polycyclic aromatic hydrocarbons (PAHs) are one of the major groups of these contaminants (Da Silva et al., 2003). PAHs constitute a diverse class of organic compounds consisting of two or more aromatic rings with various structural configurations (Prabhu and Phale, 2003). Being a derivative of benzene, PAHs are thermodynamically stable. In addition, these chemicals tend to adhere to particle surfaces, such as soils, because of their low water solubility and strong hydrophobicity, and this results in greater persistence under natural conditions. This persistence coupled with their potential carcinogenicity makes PAHs problematic environmental contaminants (Cerniglia, 1992; Sutherland, 1992). PAHs are widely found in high concentrations at many industrial sites, particularly those associated with petroleum, gas production and wood preserving industries (Wilson and Jones, 1993). 1.2 Remediation technologies Conventional techniques used for the remediation of soil polluted with organic contaminants include excavation of the contaminated soil and disposal to a landfill or capping - containment - of the contaminated areas of a site. These methods have some drawbacks. The first method simply moves the contamination elsewhere and may create significant risks in the excavation, handling and transport of hazardous material. Additionally, it is very difficult and increasingly expensive to find new landfill sites for the final disposal of the material. The cap and containment method is only an interim solution since the contamination remains on site, requiring monitoring and maintenance of the isolation barriers long into the future, with all the associated costs and potential liability. A better approach than these traditional methods is to completely destroy the pollutants, if possible, or transform them into harmless substances. Some technologies that have been used are high-temperature incineration and various types of chemical decomposition (for example, base-catalyzed dechlorination, UV oxidation). However, these methods have significant disadvantages, principally their technological complexity, high cost , and the lack of public acceptance. Bioremediation, on the contrast, is a promising option for the complete removal and destruction of contaminants. 1.3 Bioremediation of PAH contaminated soil & groundwater Bioremediation is the use of living organisms, primarily microorganisms, to degrade or detoxify hazardous wastes into harmless substances such as carbon dioxide, water and cell biomass Most PAHs are biodegradable unter natural conditions (Da Silva et al., 2003; Meysami and Baheri, 2003) and bioremediation for cleanup of PAH wastes has been extensively studied at both laboratory and commercial levels- It has been implemented at a number of contaminated sites, including the cleanup of the Exxon Valdez oil spill in Prince William Sound, Alaska in 1989, the Mega Borg spill off the Texas coast in 1990 and the Burgan Oil Field, Kuwait in 1994 (Purwaningsih, 2002). Different strategies for PAH bioremediation, such as in situ , ex situ or on site bioremediation were developed in recent years. In situ bioremediation is a technique that is applied to soil and groundwater at the site without removing the contaminated soil or groundwater, based on the provision of optimum conditions for microbiological contaminant breakdown.. Ex situ bioremediation of PAHs, on the other hand, is a technique applied to soil and groundwater which has been removed from the site via excavation (soil) or pumping (water). Hazardous contaminants are converted in controlled bioreactors into harmless compounds in an efficient manner. 1.4 Bioavailability of PAH in the subsurface Frequently, PAH contamination in the environment is occurs as contaminants that are sorbed onto soilparticles rather than in phase (NAPL, non aqueous phase liquids). It is known that the biodegradation rate of most PAHs sorbed onto soil is far lower than rates measured in solution cultures of microorganisms with pure solid pollutants (Alexander and Scow, 1989; Hamaker, 1972). It is generally believed that only that fraction of PAHs dissolved in the solution can be metabolized by microorganisms in soil. The amount of contaminant that can be readily taken up and degraded by microorganisms is defined as bioavailability (Bosma et al., 1997; Maier, 2000). Two phenomena have been suggested to cause the low bioavailability of PAHs in soil (Danielsson, 2000). The first one is strong adsorption of the contaminants to the soil constituents which then leads to very slow release rates of contaminants to the aqueous phase. Sorption is often well correlated with soil organic matter content (Means, 1980) and significantly reduces biodegradation (Manilal and Alexander, 1991). The second phenomenon is slow mass transfer of pollutants, such as pore diffusion in the soil aggregates or diffusion in the organic matter in the soil. The complex set of these physical, chemical and biological processes is schematically illustrated in Figure 1. As shown in Figure 1, biodegradation processes are taking place in the soil solution while diffusion processes occur in the narrow pores in and between soil aggregates (Danielsson, 2000). Seemingly contradictory studies can be found in the literature that indicate the rate and final extent of metabolism may be either lower or higher for sorbed PAHs by soil than those for pure PAHs (Van Loosdrecht et al., 1990). These contrasting results demonstrate that the bioavailability of organic contaminants sorbed onto soil is far from being well understood. Besides bioavailability, there are several other factors influencing the rate and extent of biodegradation of PAHs in soil including microbial population characteristics, physical and chemical properties of PAHs and environmental factors (temperature, moisture, pH, degree of contamination). Figure 1: Schematic diagram showing possible rate-limiting processes during bioremediation of hydrophobic organic contaminants in a contaminated soil-water system (not to scale) (Danielsson, 2000). 1.5 Increasing the bioavailability of PAH in soil Attempts to improve the biodegradation of PAHs in soil by increasing their bioavailability include the use of surfactants , solvents or solubility enhancers.. However, introduction of synthetic surfactant may result in the addition of one more pollutant. (Wang and Brusseau, 1993).A study conducted by Mulder et al. showed that the introduction of hydropropyl-ß-cyclodextrin (HPCD), a well-known PAH solubility enhancer, significantly increased the solubilization of PAHs although it did not improve the biodegradation rate of PAHs (Mulder et al., 1998), indicating that further research is required in order to develop a feasible and efficient remediation method. Enhancing the extent of PAHs mass transfer from the soil phase to the liquid might prove an efficient and environmentally low-risk alternative way of addressing the problem of slow PAH biodegradation in soil.

Optische Spektroskopie an Fermium (Z=100)

Im Rahmen der vorliegenden Arbeit wurde erstmals Laser-Atomspektroskopie an einem Element durchgeführt, für das bisher keine atomaren Niveaus bekannt waren. Die Experimente wurden am Element Fermium mit der Ordnungszahl Z=100 mit der Resonanzionisationsspektroskopie (RIS) in einer Puffergaszelle durchgeführt. Verwendet wurde das Isotop 255Fm mit einer Halbwertszeit von 20.1 h, das im Hochflusskernreaktor des ORNL, Oak Ridge, USA, hergestellt wurde. Die von einem elektrochemischen Filament in das Argon-Puffergas bei einer Temperatur von 960(20)°C abgedampften Fm-Atome wurden mit Lasern in einem Zweistufenprozess resonant ionisiert. Dazu wurde das Licht eines Excimerlaser gepumpten Farbstofflasers für den ersten Anregungsschritt um die Wellenlänge 400 nm durchgestimmt. Ein Teil des Excimer (XeF) Laser Pumplichtes mit den Wellenlänge 351/353 nm wurde für die nicht-resonante Ionisation verwendet. Die Ionen wurden mit Hilfe elektrischer Felder aus der optischen Zelle extrahiert und nach einem Quadrupol Massenfilter mit einem Channeltron-Detektor massenselektiv nachgewiesen. Trotz der geringen Probenmenge von 2.7 x 10^10 eingesetzten Atomen wurden zwei atomare Resonanzen bei Energien von 25099.8(2) cm-1 und 25111.8(2) cm-1 gefunden und das Sättigungsverhalten dieser Linien gemessen. Es wurde ein theoretisches Modell entwickelt, dass sowohl das spektrale Profil der sättigungsverbreiterten Linien als auch die Sättigungskurven beschreibt. Durch Anpassung an die Messdaten konnten die partiellen Übergangsraten in den 3H6 Grundzustand Aki=3.6(7) x 10^6/s und Aki=3.6(6) x 10^6/s bestimmt werden. Der Vergleich der Niveauenergien und Übergangsraten mit Multikonfigurations Dirac-Fock Rechnungen legt die spektroskopische Klassifizierung der beobachteten Niveaus als 5f12 7s7p 5I6 und 5G6 Terme nahe. Weiterhin wurde ein Übergang bei 25740 cm-1 gefunden, der aufgrund der beobachteten Linienbreite von 1000 GHz als Rydbergzustand Zustand mit der Niveauenergie 51480 cm-1 interpretiert wurde und über einen Zweiphotonen Prozess angeregt werden kann. Basierend auf dieser Annahme wurde die Obergrenze für die Ionisationsenergie IP = 52140 cm-1 = 6.5 eV abgeschätzt. In den Messungen wurden Verschiebungen in den Zeitverteilungsspektren zwischen den mono-atomaren Ionen Fm+ und Cf+ und dem Molekül-Ion UO+ festgestellt und auf Driftzeitunterschiede im elektrischen Feld der gasgefüllten optischen Zelle zurückgeführt. Unter einfachen Modellannahmen wurde daraus auf die relativen Unterschiede Delta_r(Fm+,Cf+)/r(Cf+) ˜ -0.2 % und Delta_r(UO+,Cf+)/r(Cf+) ˜ 20 % in den Ionenradien geschlossen. Über die Bestimmung der Abnahme der Fm-a Aktivität des Filamentes auf der einen Seite und die Messung der Resonanzzählrate auf der anderen Seite, wurde die Nachweiseffizienz der Apparatur zu 4.5(3) x 10^-4 bestimmt. Die Nachweisapparatur wurde mit dem Ziel weiterentwickelt, Laserspektroskopie am Isotop 251Fm durchzuführen, das über die Reaktion 249Cf(a,2n)251Fm direkt in der optischen Zelle erzeugt werden soll. Das Verfahren wurde am chemischen Homolog Erbium getestet. Dabei wurde das Isotop 163Er über die Reaktion 161Dy(a,2n)163Er erzeugt und nach Resonanzionisation nachgewiesen. Die Nachweiseffizienz der Methode wurde zu 1 x 10^-4 bestimmt.

Harmful algae and their potential impacts on the coastal ecosystem: growth and toxin production dynamics

The main goal of the present thesis was to study some harmful algal species which cause blooms in Italian coastal waters, leading to consequences for human health, coastal ecosystem, fishery and tourism. In particular, in the first part of this thesis the toxicity of Adriatic strains of the raphidophyte Fibrocapsa japonica was investigated. Despite several hypotheses have been proposed for the toxic mechanism of the raphidophytes, especially for the species Chattonella antiqua and C. marina, which have been studied more extensively, just a few studies on the toxic effects of these species for different organisms were reported. Moreover, a careful reading of the literature evidenced as any ichthyotoxic events reported worldwide can be linked to F. japonica blooms. Although recently several studies were performed on F. japonica strains from the USA, Japan, Australia, New Zealand, the Netherlands, Germany, and France in order to characterize their growth and toxicity features, the work reported in this thesis results one of the first investigation on the toxic effects of F. japonica for different organisms, such as bacteria, crustaceans and fish. Mortality effects, together with haemolysis of fish erythrocytes, probably due to the relatively high amount of PUFAs produced by this species, were observed. Mortality for fish, however, was reported only at a high cell density and after a long exposition period (9-10 days); moreover a significant increase of H2O2 obtained in the tanks where sea basses were exposed to F. japonica was also relevant. This result may justify the absence of ichthyotoxic events in the Italian coasts, despite F. japonica blooms detected in these areas were characterized by high cell densities. This work reports also a first complete characterization of the fatty acids produced and extracellularly released by the Adriatic F. japonica, and results were also compared with the fatty acid profile of other strains. The absence of known brevetoxins in F. japonica algal extracts was also highlighted, leading to the hypothesis that the toxicity of F. japonica may be due to a synergic effect of PUFAs and ROS. Another microalgae that was studied in this thesis is the benthic dinoflagellate Ostreopsis cf. ovata. This species was investigated with the aim to investigate the effect of environmental parameters on its growth and toxicity. O. cf. ovata, in fact, shows different blooming periods along the Italian coasts and even the reported toxic effects are variable. The results of this work confirmed the high variability in the growth dynamic and toxin content of several Italian strains which were isolated in recent years along the Adriatic and Tyrrhenian Seas. Moreover, the effects of temperature and salinity on the behaviour of the different isolates are in good agreement with the results obtained from field surveys, which evidence as the environmental parameters are important factors modulating O. cf. ovata proliferation. Another relevant result that was highlighted is the anomaly in the production of palytoxin-like compounds reported by one of the studied isolate, in particular the one isolated in 2008 in Ancona (Adriatic Sea). Only this strain reported the absence of two (ovatoxin-b and –c) of the five ovatoxins so far known in the toxin profile and a different relative abundance of the other toxins. The last aspect that was studied in this thesis regards the toxin biosythesis. In fact, toxins produced (palytoxin-like compounds) or supposed to be produced (brevetoxin-like compounds) by O. cf. ovata and F. japonica, respectively, are polyketides, which are highly oxygenated compounds synthesized by complex enzymes known as polyketide synthase (PKS) enzymes. These enzymes are multi-domain complexes that structurally and functionally resemble the fatty acid synthases (FASs). This work reports the first study of PKS proteins in the dinoflagellates O. cf. ovata, C. monotis and in the raphidophyte F. japonica. For the first time some PKSs were identified in these species, confirming the presence of PKS proteins predicted by the in silico translation of the transcripts found in K. brevis also in other species. The identification of O. cf. ovata PKSs and the localization of the palytoxin-like compounds produced by this dinoflagellate in a similar location (chloroplast) as that observed for other dinoflagellate and cyanobacterial toxins provides some indication that these proteins may be involved in polyketide biosynthesis. However, their potential function as fatty acid synthases cannot be ruled out, as plant fatty acid synthesis also occurs within chloroplasts. This last hypothesis is also supported by the fact that in all the investigated species, and in particular in F. japonica, PKS proteins were present. Therefore, these results provide an important contribution to the study of the polyketides and of the involvement of PKS proteins in the toxin biosynthesis.

Nanoscopic studies of conjugated polymer blends by (electric) scanning probe microscopy

Conjugated polymers and conjugated polymer blends have attracted great interest due to their potential applications in biosensors and organic electronics. The sub-100 nm morphology of these materials is known to heavily influence their electromechanical properties and the performance of devices they are part of. Electromechanical properties include charge injection, transport, recombination, and trapping, the phase behavior and the mechanical robustness of polymers and blends. Electrical scanning probe microscopy techniques are ideal tools to measure simultaneously electric (conductivity and surface potential) and dielectric (dielectric constant) properties, surface morphology, and mechanical properties of thin films of conjugated polymers and their blends.rnIn this thesis, I first present a combined topography, Kelvin probe force microscopy (KPFM), and scanning conductive torsion mode microscopy (SCTMM) study on a gold/polystyrene model system. This system is a mimic for conjugated polymer blends where conductive domains (gold nanoparticles) are embedded in a non-conductive matrix (polystyrene film), like for polypyrrole:polystyrene sulfonate (PPy:PSS), and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS). I controlled the nanoscale morphology of the model by varying the distribution of gold nanoparticles in the polystyrene films. I studied the influence of different morphologies on the surface potential measured by KPFM and on the conductivity measured by SCTMM. By the knowledge I gained from analyzing the data of the model system I was able to predict the nanostructure of a homemade PPy:PSS blend.rnThe morphologic, electric, and dielectric properties of water based conjugated polymer blends, e.g. PPy:PSS or PEDOT:PSS, are known to be influenced by their water content. These properties also influence the macroscopic performance when the polymer blends are employed in a device. In the second part I therefore present an in situ humidity-dependence study on PPy:PSS films spin-coated and drop-coated on hydrophobic highly ordered pyrolytic graphite substrates by KPFM. I additionally used a particular KPFM mode that detects the second harmonic electrostatic force. With this, I obtained images of dielectric constants of samples. Upon increasing relative humidity, the surface morphology and composition of the films changed. I also observed that relative humidity affected thermally unannealed and annealed PPy:PSS films differently. rnThe conductivity of a conjugated polymer may change once it is embedded in a non-conductive matrix, like for PPy embedded in PSS. To measure the conductivity of single conjugated polymer particles, in the third part, I present a direct method based on microscopic four-point probes. I started with metal core-shell and metal bulk particles as models, and measured their conductivities. The study could be extended to measure conductivity of single PPy particles (core-shell and bulk) with a diameter of a few micrometers.

Power Converters and Electric Drives for Smart Grid Applications

The present dissertation aims to explore, theoretically and experimentally, the problems and the potential advantages of different types of power converters for “Smart Grid” applications, with particular emphasis on multi-level architectures, which are attracting a rising interest even for industrial requests. The models of the main multilevel architectures (Diode-Clamped and Cascaded) are shown. The best suited modulation strategies to function as a network interface are identified. In particular, the close correlation between PWM (Pulse Width Modulation) approach and SVM (Space Vector Modulation) approach is highlighted. An innovative multilevel topology called MMC (Modular Multilevel Converter) is investigated, and the single-phase, three-phase and "back to back" configurations are analyzed. Specific control techniques that can manage, in an appropriate way, the charge level of the numerous capacitors and handle the power flow in a flexible way are defined and experimentally validated. Another converter that is attracting interest in “Power Conditioning Systems” field is the “Matrix Converter”. Even in this architecture, the output voltage is multilevel. It offers an high quality input current, a bidirectional power flow and has the possibility to control the input power factor (i.e. possibility to participate to active and reactive power regulations). The implemented control system, that allows fast data acquisition for diagnostic purposes, is described and experimentally verified.

Potential vorticity and moisture in extratropical cyclones : climatology and sensitivity experiments

The development of extratropical cyclones can be seen as an interplay of three positive potential vorticity (PV) anomalies: an upper-level stratospheric intrusion, low-tropospheric diabatically produced PV, and a warm anomaly at the surface acting as a surrogate PV anomaly. In the mature stage they become vertically aligned and form a “PV tower” associated with strong cyclonic circulation. This paradigm of extratropical cyclone development provides the basis of this thesis, which will use a climatological dataset and numerical model experiments to investigate the amplitude of the three anomalies and the processes leading in particular to the formation of the diabatically produced low-tropospheric PV anomaly.rnrnThe first part of this study, based on the interim ECMWF Re-Analysis (ERA-Interim) dataset, quantifies the amplitude of the three PV anomalies in mature extratropical cyclones in different regions in the Northern Hemisphere on a climatological basis. A tracking algorithm is applied to sea level pressure (SLP) fields to identify cyclone tracks. Surface potential temperature anomalies ∆θ and vertical profiles of PV anomalies ∆PV are calculated at the time of the cyclones’ minimum SLP and during the intensification phase 24 hours before in a vertical cylinder with a radius of 200 km around the surface cyclone center. To compare the characteristics of the cyclones, they are grouped according to their location (8 regions) and intensity, where the central SLP is used as a measure of intensity. Composites of ∆PV profiles and ∆θ are calculated for each region and intensity class at the time of minimum SLP and during the cyclone intensification phase.rnrnDuring the cyclones’ development stage the amplitudes of all three anomalies increase on average. In the mature stage all three anomalies are typically larger for intense than for weak winter cyclones [e.g., 0.6 versus 0.2 potential vorticity units (PVU) at lower levels, and 1.5 versus 0.5 PVU at upper levels].rnThe regional variability of the cyclones’ vertical structure and the profile evolution is prominent (cyclones in some regions are more sensitive to the amplitude of a particular anomaly than in other regions). Values of ∆θ and low-level ∆PV are on average larger in the western parts of the oceans than in the eastern parts. In addition, a large seasonal variability can be identified, with fewer and weaker cyclones especially in the summer, associated with higher low-tropospheric PV values, but also with a higher tropopause and much weaker surface potential temperature anomalies (compared to winter cyclones).rnrnIn the second part, we were interested in the diabatic low-level part of PV towers. Evaporative sources were identified of moisture that was involved in PV production through condensation. Lagrangian backward trajectories were calculated from the region with high PV values at low-levels in the cyclones. PV production regions were identified along these trajectories and from these regions a new set of backward trajectories was calculated and moisture uptakes were traced along them. The main contribution from surface evaporation to the specific humidity of the trajectories is collected 12-72 hours prior to therntime of PV production. The uptake region for weaker cyclones with less PV in the centre is typically more localized with reduced uptake values compared to intense cyclones. However, in a qualitative sense uptakes and other variables along single trajectories do not vary much between cyclones of different intensity in different regions.rnrnA sensitivity study with the COSMO model comprises the last part of this work. The study aims at investigating the influence of synthetic moisture modification in the cyclone environment in different stages of its development. Moisture was eliminated in three regions, which were identified as important moisture source regions for PV production. Moisture suppression affected the cyclone the most in its early phase. It led to cyclolysis shortly after its genesis. Nevertheles, a new cyclone formed on the other side of a dry box and developed relatively quickly. Also in other experiments, moisture elimination led to strong intensity reduction of the surface cyclone, limited upper-level development, and delayed or missing interaction between the two.rnrnIn summary, this thesis provides novel insight into the structure of different intensity categories of extratropical cyclones from a PV perspective, which corroborates the findings from a series of previous case studies. It reveals that all three PV anomalies are typically enhanced for more intense cyclones, with important regional differences concerning the relative amplitude of the three anomalies. The moisture source analysis is the first of this kind to study the evaporation-condensation cycle related to the intensification of extratropical cyclones. Interestingly, most of the evaporation occurs during the 3 days prior to the time of maximum cyclone intensity and typically extends over fairly large areas along the track of the cyclone. The numerical model case study complements this analysis by analyzing the impact of regionally confined moisture sources for the evolution of the cyclone.

An integrated nano-scale approach to profile miRNAs in limited clinical samples

Profiling miRNA expression in cells that directly contribute to human disease pathogenesis is likely to aid the discovery of novel drug targets and biomarkers. However, tissue heterogeneity and the limited amount of human diseased tissue available for research purposes present fundamental difficulties that often constrain the scope and potential of such studies. We established a flow cytometry-based method for isolating pure populations of pathogenic T cells from bronchial biopsy samples of asthma patients, and optimized a high-throughput nano-scale qRT-PCR method capable of accurately measuring 96 miRNAs in as little as 100 cells. Comparison of circulating and airway T cells from healthy and asthmatic subjects revealed asthma-associated and tissue-specific miRNA expression patterns. These results establish the feasibility and utility of investigating miRNA expression in small populations of cells involved in asthma pathogenesis, and set a precedent for application of our nano-scale approach in other human diseases. The microarray data from this study (Figure 7) has been submitted to the NCBI Gene Expression Omnibus (GEO; http://ncbi.nlm.nih.gov/geo) under accession no. GSE31030.

Prioritization of Potential Mates' History of Sexual Fidelity During a Conjoint Ranking Task

This series of studies is the first to use conjoint analysis to examine how individuals make trade-offs during mate selection when provided information about a partner's history of sexual infidelity. Across three studies, participants ranked profiles of potential mates, with each profile varying across five attributes: financial stability, physical attractiveness, sexual fidelity, emotional investment, and similarity. They also rated each attribute separately for importance in an ideal mate. Overall, we found that for a long-term mate, participants prioritized a potential partner's history of sexual fidelity over other attributes when profiles were ranked conjointly. For a short-term mate, sexual fidelity, physical attractiveness, and financial stability were equally important, and each was more important than emotional investment and similarity. These patterns contrast with participants' self-reported importance ratings of each individual attribute. Our results are interpreted within the context of previous literature examining how making trade-offs affect mate selection.

Bacterial profile and burden of periodontal infection in subjects with a diagnosis of acute coronary syndrome

BACKGROUND: Periodontitis has been identified as a potential risk factor in cardiovascular diseases. It is possible that the stimulation of host responses to oral infections may result in vascular damage and the inducement of blood clotting. The aim of this study was to assess the role of periodontal infection and bacterial burden as an explanatory variable to the activation of the inflammatory process leading to acute coronary syndrome (ACS). METHODS: A total of 161 consecutive surviving cases admitted with a diagnosis of ACS and 161 control subjects, matched with cases according to their gender, socioeconomic level, and smoking status, were studied. Serum white blood cell (WBC) counts, high- and low-density lipoprotein (HDL/LDL) levels, high-sensitivity C-reactive protein (hsC-rp) levels, and clinical periodontal routine parameters were studied. The subgingival pathogens were assayed by the checkerboard DNA-DNA hybridization method. RESULTS: Total oral bacterial load was higher in the subjects with ACS (mean difference: 17.4x10(5); SD: 10.8; 95% confidence interval [CI]: 4.2 to 17.4; P<0.001), and significant for 26 of 40 species including Porphyromonas gingivalis, Tannerella forsythensis, and Treponema denticola. Serum WBC counts, hsC-rp levels, Streptococcus intermedius, and Streptococcus sanguis, were explanatory factors to acute coronary syndrome status (Nagelkerke r2=0.49). CONCLUSION: The oral bacterial load of S. intermedius, S. sanguis, Streptococcus anginosus, T. forsythensis, T. denticola, and P. gingivalis may be concomitant risk factors in the development of ACS.