Anisotropic Brownian motion in ordered phases of DNA fragments

Using Fluorescence Recovery After Photobleaching, we investigate the Brownian motion of DNA rod-like fragments in two distinct anisotropic phases with a local nematic symmetry. The height of the measurement volume ensures the averaging of the anisotropy of the in-plane diffusive motion parallel or perpendicular to the local nematic director in aligned domains. Still, as shown in using a model specifically designed to handle such a situation and predicting a non-Gaussian shape for the bleached spot as fluorescence recovery proceeds, the two distinct diffusion coefficients of the DNA particles can be retrieved from data analysis. In the first system investigated (a ternary DNA-lipid lamellar complex), the magnitude and anisotropy of the diffusion coefficient of the DNA fragments confined by the lipid bilayers are obtained for the first time. In the second, binary DNA-solvent system, the magnitude of the diffusion coefficient is found to decrease markedly as DNA concentration is increased from isotropic to cholesteric phase. In addition, the diffusion coefficient anisotropy measured within cholesteric domains in the phase coexistence region increases with concentration, and eventually reaches a high value in the cholesteric phase.

Field and Laboratory Studies Provide Insights into the Meaning of Day-Time Activity in a Subterranean Rodent (Ctenomys aff. knighti), the Tuco-Tuco

South American subterranean rodents (Ctenomys aff. knighti), commonly known as tuco-tucos, display nocturnal, wheel-running behavior under light-dark (LD) conditions, and free-running periods >24 h in constant darkness (DD). However, several reports in the field suggested that a substantial amount of activity occurs during daylight hours, leading us to question whether circadian entrainment in the laboratory accurately reflects behavior in natural conditions. We compared circadian patterns of locomotor activity in DD of animals previously entrained to full laboratory LD cycles (LD12:12) with those of animals that were trapped directly from the field. In both cases, activity onsets in DD immediately reflected the previous dark onset or sundown. Furthermore, freerunning periods upon release into DD were close to 24 h indicating aftereffects of prior entrainment, similarly in both conditions. No difference was detected in the phase of activity measured with and without access to a running wheel. However, when individuals were observed continuously during daylight hours in a semi-natural enclosure, they emerged above-ground on a daily basis. These day-time activities consisted of foraging and burrow maintenance, suggesting that the designation of this species as nocturnal might be inaccurate in the field. Our study of a solitary subterranean species suggests that the circadian clock is entrained similarly under field and laboratory conditions and that day-time activity expressed only in the field is required for foraging and may not be time-dictated by the circadian pacemaker.

IL-5 and IL-17A are critical for the chronic IgE response and differentiation of long-lived antibody-secreting cells in inflamed tissues

Prolonged survival of long-lived antibody-secreting cells in the BM has been implicated as a key component of long-term humoral immunity. The current study was designed to uncover the extrinsic signals required for the generation and maintenance of ASC in several niches (peritoneum, spleen and bone-marrow). Our results show that protein mixture of the Thalassophryne nattereri venom induced a chronic Th2 humoral response that is characterized by splenic hyperplasia with GC formation and venom retention by follicular DCs. Retention of B1a in the BM were observed. In the late phase (120 d) of chronic venom-response the largest pool of ASC into the peritoneal cavity consisted of B220(neg)CD43(high) phenotype; the largest pool of ASC into spleen was constituted by B220 positive cells (B220(high) and B220(low)), whereas the largest pool of ASC into in the BM was constituted by the B220(high)CD43(low) phenotype; and finally, terminally differentiated cells (B220(neg)CD43(high)) were only maintained in the inflamed peritoneal cavity in late phase. After 120 d a sustained production of cytokines (KC, IL-5, TNF-alpha, IL-6, IL-17A and IL-23) and leukocytes recruitment (eosinophils, mast cells, and neutrophils) were induced. IL-5- and IL-17A-producing CD4+ CD44+ CD40L+ Ly6C+ effector memory T cells were also observed in peritoneal cavity. Finally, treatment of venom-mice with anti-IL-5- and anti-IL17A-neutralizing mAbs abolished the synthesis of specific IgE, without modifying the splenic hyperplasia or GC formation. In addition, IL-5 and IL-17A negatively regulated the expansion of B1a in peritoneal cavity and BM, and promoted the differentiation of these cells in spleen. And more, IL-5 and IL-17A are sufficient for the generation of ASC B220(neg) in the peritoneal cavity and negatively regulate the number of ASC B220(Pos), confirming that the hierarchical process of ASC differentiation triggered by venom needs the signal derived from IL-5 and IL-17A. (C) 2012 Elsevier Ltd. All rights reserved.

Molecular measurement of BCR-ABL transcript variations in chronic myeloid leukemia patients in cytogenetic remission

Abstract Background The monitoring of BCR-ABL transcript levels by real-time quantitative polymerase chain reaction (RT-qPCR) has become important to assess minimal residual disease (MRD) and standard of care in the treatment of chronic myeloid leukemia (CML). In this study, we performed a prospective, sequential analysis using RT-qPCR monitoring of BCR-ABL gene rearrangements in blood samples from 91 CML patients in chronic phase (CP) who achieved complete cytogenetic remission (CCyR) and major molecular remission (MMR) throughout imatinib treatment. Methods The absolute level of BCR-ABL transcript from peripheral blood was serially measured every 4 to 12 weeks by RT-qPCR. Only level variations > 0.5%, according to the international scale, was considered positive. Sequential cytogenetic analysis was also performed in bone marrow samples from all patients using standard protocols. Results Based on sequential analysis of BCR-ABL transcripts, the 91 patients were divided into three categories: (A) 57 (62.6%) had no variation on sequential analysis; (B) 30 (32.9%) had a single positive variation result obtained in a single sample; and (C) 4 (4.39%) had variations of BCR-ABL transcripts in at least two consecutive samples. Of the 34 patients who had elevated levels of transcripts (group B and C), 19 (55.8%) had a < 1% of BCR-ABL/BCR ratio, 13 (38.2%) patients had a 1% to 10% increase and 2 patients had a >10% increase of RT-qPCR. The last two patients had lost a CCyR, and none of them showed mutations in the ABL gene. Transient cytogenetic alterations in Ph-negative cells were observed in five (5.5%) patients, and none of whom lost CCyR. Conclusions Despite an increase levels of BCR-ABL/BCR ratio variations by RT-qPCR, the majority of CML patients with MMR remained in CCyR. Thus, such single variations should neither be considered predictive of subsequent failure and nor an indication for altering imatinib dose or switching to second generation therapy. Changing of imatinib on the basis of BCR-ABL/BCR% sustained increase and mutational studies is a prudent approach for preserving other therapeutic options in imatinib-resistant patients.

In situ effect of chewing gum containing CPP–ACP on the mineral precipitation of eroded bovine enamel

Objectives: Stimulation of salivary flow is considered a preventive strategy for dental erosion. Alternatively, products containing calcium phosphate, such as a complex of casein phosphopeptide–amorphous calcium phosphate (CPP–ACP), have also been tested against dental erosion. Therefore, this in situ study analyzed the effect of chewing gum containing CPP–ACP on the mineral precipitation of initial bovine enamel erosion lesions. Methods: Twelve healthy adult subjects wore palatal appliances with two eroded bovine enamel samples. The erosion lesions were produced by immersion in 0.1% citric acid (pH 2.5) for 7 min. During three experimental crossover in situ phases (1 day each), the subjects chewed a type of gum, 3 times for 30 min, in each phase: with CPP–ACP (trident total), without CPP–ACP (trident), and no chewing gum (control). The Knoop surface microhardness was measured at baseline, after erosion in vitro and the mineral precipitation in situ. The differences in the degree of mineral precipitation were analyzed using repeated measures (RM-) ANOVA and post hoc Tukey’s test ( p < 0.05). Results: Significant differences were found among the remineralizing treatments ( p < 0.0001). Chewing gum (19% of microhardness recovery) improved the mineral precipitation compared to control (10%) and the addition of CPP–ACP into the gum promoted the best mineral precipitation effect (30%). Conclusions: Under this protocol, CPP–ACP chewing gum improved the mineral precipitation of eroded enamel. Clinical significance: Since the prevalence of dental erosion is steadily increasing, CPP–ACP chewing gum might be an important strategy to reduce th eprogression of initial erosion lesions.

Computational chemistry studies of UV induced processes in human skin

This thesis presents and uses the techniques of computational chemistry to explore two different processes induced in human skin by ultraviolet light. The first is the transformation of urocanic acid into a immunosuppressing agent, and the other is the enzymatic action of the 8-oxoguanine glycosylase enzyme. The photochemistry of urocanic acid is investigated by time-dependent density functional theory. Vertical absorption spectra of the molecule in different forms and environments is assigned and candidate states for the photochemistry at different wavelengths are identified. Molecular dynamics simulations of urocanic acid in gas phase and aqueous solution reveals considerable flexibility under experimental conditions, particularly for for the cis isomer where competition between intra- and inter-molecular interactions increases flexibility. A model to explain the observed gas phase photochemistry of urocanic acid is developed and it is shown that a reinterpretation in terms of a mixture between isomers significantly enhances the agreement between theory and experiment , and resolves several peculiarities in the spectrum. A model for the photochemistry in the aqueous phase of urocanic acid is then developed, in which two excited states governs the efficiency of photoisomerization. The point of entrance into a conical intersection seam is shown to explain the wavelength dependence of photoisomerization quantum yield. Finally some mechanistic aspects of the DNA repair enzyme 8-oxoguanine glycosylase is investigated with density functional theory. It is found that the critical amino acid of the active site can provide catalytic power in several different manners, and that a recent proposal involving a SN1 type of mechanism seems the most efficient one.

Structural changes and dynamic behaviour of vanadium oxide-based catalysts for gas-phase selective oxidations

Selective oxidation is one of the simplest functionalization methods and essentially all monomers used in manufacturing artificial fibers and plastics are obtained by catalytic oxidation processes. Formally, oxidation is considered as an increase in the oxidation number of the carbon atoms, then reactions such as dehydrogenation, ammoxidation, cyclization or chlorination are all oxidation reactions. In this field, most of processes for the synthesis of important chemicals used vanadium oxide-based catalysts. These catalytic systems are used either in the form of multicomponent mixed oxides and oxysalts, e.g., in the oxidation of n-butane (V/P/O) and of benzene (supported V/Mo/O) to maleic anhydride, or in the form of supported metal oxide, e.g., in the manufacture of phthalic anhydride by o-xylene oxidation, of sulphuric acid by oxidation of SO2, in the reduction of NOx with ammonia and in the ammoxidation of alkyl aromatics. In addition, supported vanadia catalysts have also been investigated for the oxidative dehydrogenation of alkanes to olefins , oxidation of pentane to maleic anhydride and the selective oxidation of methanol to formaldehyde or methyl formate [1]. During my PhD I focused my work on two gas phase selective oxidation reactions. The work was done at the Department of Industrial Chemistry and Materials (University of Bologna) in collaboration with Polynt SpA. Polynt is a leader company in the development, production and marketing of catalysts for gas-phase oxidation. In particular, I studied the catalytic system for n-butane oxidation to maleic anhydride (fluid bed technology) and for o-xylene oxidation to phthalic anhydride. Both reactions are catalyzed by systems based on vanadium, but catalysts are completely different. Part A is dedicated to the study of V/P/O catalyst for n-butane selective oxidation, while in the Part B the results of an investigation on TiO2-supported V2O5, catalyst for o-xylene oxidation are showed. In Part A, a general introduction about the importance of maleic anhydride, its uses, the industrial processes and the catalytic system are reported. The reaction is the only industrial direct oxidation of paraffins to a chemical intermediate. It is produced by n-butane oxidation either using fixed bed and fluid bed technology; in both cases the catalyst is the vanadyl pyrophosphate (VPP). Notwithstanding the good performances, the yield value didn’t exceed 60% and the system is continuously studied to improve activity and selectivity. The main open problem is the understanding of the real active phase working under reaction conditions. Several articles deal with the role of different crystalline and/or amorphous vanadium/phosphorous (VPO) compounds. In all cases, bulk VPP is assumed to constitute the core of the active phase, while two different hypotheses have been formulated concerning the catalytic surface. In one case the development of surface amorphous layers that play a direct role in the reaction is described, in the second case specific planes of crystalline VPP are assumed to contribute to the reaction pattern, and the redox process occurs reversibly between VPP and VOPO4. Both hypotheses are supported also by in-situ characterization techniques, but the experiments were performed with different catalysts and probably under slightly different working conditions. Due to complexity of the system, these differences could be the cause of the contradictions present in literature. Supposing that a key role could be played by P/V ratio, I prepared, characterized and tested two samples with different P/V ratio. Transformation occurring on catalytic surfaces under different conditions of temperature and gas-phase composition were studied by means of in-situ Raman spectroscopy, trying to investigate the changes that VPP undergoes during reaction. The goal is to understand which kind of compound constituting the catalyst surface is the most active and selective for butane oxidation reaction, and also which features the catalyst should possess to ensure the development of this surface (e.g. catalyst composition). On the basis of results from this study, it could be possible to project a new catalyst more active and selective with respect to the present ones. In fact, the second topic investigated is the possibility to reproduce the surface active layer of VPP onto a support. In general, supportation is a way to improve mechanical features of the catalysts and to overcome problems such as possible development of local hot spot temperatures, which could cause a decrease of selectivity at high conversion, and high costs of catalyst. In literature it is possible to find different works dealing with the development of supported catalysts, but in general intrinsic characteristics of VPP are worsened due to the chemical interaction between active phase and support. Moreover all these works deal with the supportation of VPP; on the contrary, my work is an attempt to build-up a V/P/O active layer on the surface of a zirconia support by thermal treatment of a precursor obtained by impregnation of a V5+ salt and of H3PO4. In-situ Raman analysis during the thermal treatment, as well as reactivity tests are used to investigate the parameters that may influence the generation of the active phase. Part B is devoted to the study of o-xylene oxidation of phthalic anhydride; industrially, the reaction is carried out in gas-phase using as catalysts a supported system formed by V2O5 on TiO2. The V/Ti/O system is quite complex; different vanadium species could be present on the titania surface, as a function of the vanadium content and of the titania surface area: (i) V species which is chemically bound to the support via oxo bridges (isolated V in octahedral or tetrahedral coordination, depending on the hydration degree), (ii) a polymeric species spread over titania, and (iii) bulk vanadium oxide, either amorphous or crystalline. The different species could have different catalytic properties therefore changing the relative amount of V species can be a way to optimize the catalytic performances of the system. For this reason, samples containing increasing amount of vanadium were prepared and tested in the oxidation of o-xylene, with the aim of find a correlations between V/Ti/O catalytic activity and the amount of the different vanadium species. The second part deals with the role of a gas-phase promoter. Catalytic surface can change under working conditions; the high temperatures and a different gas-phase composition could have an effect also on the formation of different V species. Furthermore, in the industrial practice, the vanadium oxide-based catalysts need the addition of gas-phase promoters in the feed stream, that although do not have a direct role in the reaction stoichiometry, when present leads to considerable improvement of catalytic performance. Starting point of my investigation is the possibility that steam, a component always present in oxidation reactions environment, could cause changes in the nature of catalytic surface under reaction conditions. For this reason, the dynamic phenomena occurring at the surface of a 7wt% V2O5 on TiO2 catalyst in the presence of steam is investigated by means of Raman spectroscopy. Moreover a correlation between the amount of the different vanadium species and catalytic performances have been searched. Finally, the role of dopants has been studied. The industrial V/Ti/O system contains several dopants; the nature and the relative amount of promoters may vary depending on catalyst supplier and on the technology employed for the process, either a single-bed or a multi-layer catalytic fixed-bed. Promoters have a quite remarkable effect on both activity and selectivity to phthalic anhydride. Their role is crucial, and the proper control of the relative amount of each component is fundamental for the process performance. Furthermore, it can not be excluded that the same promoter may play different role depending on reaction conditions (T, composition of gas phase..). The reaction network of phthalic anhydride formation is very complex and includes several parallel and consecutive reactions; for this reason a proper understanding of the role of each dopant cannot be separated from the analysis of the reaction scheme. One of the most important promoters at industrial level, which is always present in the catalytic formulations is Cs. It is known that Cs plays an important role on selectivity to phthalic anhydride, but the reasons of this phenomenon are not really clear. Therefore the effect of Cs on the reaction scheme has been investigated at two different temperature with the aim of evidencing in which step of the reaction network this promoter plays its role.

Analisi biomeccanica del MAWASHI GERI JODAN in cinture nere di Karate: modelli tecnici a confronto

In martial arts there are several ways to perform the turning kick . Following the martial arts or different learning models many types of kicks take shape. Mawashi geri is the karate turning kick. At the moment there are two models of mawashi geri, one comes from the traditional karate (OLD), and the other newer (NEW), who agrees to the change of the rules of W.K.F. (World Karate Federation) happened in 2000 (Macan J. et all 2006) . In this study we are focus on the differences about two models the mawashi geri jodan of karate. The purpose of this study is to analyse cinematic and kinetic parameters of mawashi geri jodan. Timing of the striking and supporting leg actions were also evaluated A Vicon system 460 IR with 6 cameras at sample frequency of 200 Hz was used. 37 reflective markers have been set on the skin of the subjects following the “PlugInGait-total body model”. The participants performed five repetitions of mawashi geri jodan at maximum rapidity with their dominant leg against a ball suspended in front of them placed at ear height. Fourteen skilled subjects (mean level black belt 1,7 dan; age 20,9±4,8 yrs; height 171,4±7,3 cm; weight 60,9±10,2 Kg) practicing karate have been split in two group through the hierarchical cluster analysis following their technical characteristics. By means of the Mann Whitney-U test (Spss-package) the differences between the two groups were verified in preparatory and execution phase. Kicking knee at start, kicking hip and knee at take-off were different between the two groups (p < 0,05). Striking hip flexion during the spin of the supporting foot was different between the two groups (p < 0,05). Peak angular velocity of hip flexion were different between the two groups (p < 0,05). Groups showed differences also in timing of the supporting spin movement. While Old group spin the supporting foot at 30% of the trial, instead New start spinning at 44% of the trial. Old group showed a greater supporting foot spin than New (Old 110° Vs New 82°). Abduction values didn’t show any differences between the two groups. At the hit has been evaluated a 120° of double hips abduction, for the entire sample. Striking knee extension happened for everybody after the kicking hip flexion and confirm the proximal-distal action of the striking leg (Sorensen H. 1996). In contrast with Pearson J.N. 1997 and Landeo R 2007, peak velocity of the striking foot is not useful to describe kick performance because affected by the stature. Two groups are different either in preparatory phase or in execution phase. The body is set in difference manner already before the take-off of the kicking foot. The groups differ for the timing of the supporting foot action Trainer should pay attention to starting posture and on abduction capacities of the athletes.

New path integral simulation algorithms and their application to creep in the quantum sine-Gordon chain

A path integral simulation algorithm which includes a higher-order Trotter approximation (HOA)is analyzed and compared to an approach which includes the correct quantum mechanical pair interaction (effective Propagator (EPr)). It is found that the HOA algorithmconverges to the quantum limit with increasing Trotter number P as P^{-4}, while the EPr algorithm converges as P^{-2}.The convergence rate of the HOA algorithm is analyzed for various physical systemssuch as a harmonic chain,a particle in a double-well potential, gaseous argon, gaseous helium and crystalline argon. A new expression for the estimator for the pair correlation function in the HOA algorithm is derived. A new path integral algorithm, the hybrid algorithm, is developed.It combines an exact treatment of the quadratic part of the Hamiltonian and thehigher-order Trotter expansion techniques.For the discrete quantum sine-Gordon chain (DQSGC), it is shown that this algorithm works more efficiently than all other improved path integral algorithms discussed in this work. The new simulation techniques developed in this work allow the analysis of theDQSGC and disordered model systems in the highly quantum mechanical regime using path integral molecular dynamics (PIMD)and adiabatic centroid path integral molecular dynamics (ACPIMD).The ground state phonon dispersion relation is calculated for the DQSGC by the ACPIMD method.It is found that the excitation gap at zero wave vector is reduced by quantum fluctuations. Two different phases exist: One phase with a finite excitation gap at zero wave vector, and a gapless phase where the excitation gap vanishes.The reaction of the DQSGC to an external driving force is analyzed at T=0.In the gapless phase the system creeps if a small force is applied, and in the phase with a gap the system is pinned. At a critical force, the systems undergo a depinning transition in both phases and flow is induced. The analysis of the DQSGC is extended to models with disordered substrate potentials. Three different cases are analyzed: Disordered substrate potentials with roughness exponent H=0, H=1/2,and a model with disordered bond length. For all models, the ground state phonon dispersion relation is calculated.

Idrogenazione selettiva del 5-idrossimetilfurfurale in fase omogenea: studio dell'attività catalitica del complesso di Shvo

Il presente lavoro di tesi è frutto di una collaborazione fra il Dipartimento di Chimica Fisica ed Inorganica (gruppo del Prof. Valerio Zanotti – Mattia Vaccari, Dr. Rita Mazzoni) ed il Dipartimento di Chimica Industriale e dei Materiali (gruppo del Prof. Angelo Vaccari – Dr. Thomas Pasini, Dr. Stefania Albonetti, Prof. Fabrizio Cavani) e si inserisce il un progetto volto a valutare l’attività e la selettività del catalizzatore di idrogenazione di Shvo 1, verso l’idrogenazione selettiva del doppio legame polare del 5-idrossimetilfurfurale (HMF) in fase omogenea. L’HMF è un composto di natura organica facilmente ottenibile dalle biomasse, il quale può essere impiegato come building block per ottenere prodotti ad alto valore aggiunto per la chimica fine o additivi per biocarburanti aventi un elevato potere calorifico. In particolare la nostra attenzione si è rivolta alla produzione del 2,5-diidrossimetilfurano (BHMF), un importante building block per la produzione di polimeri e schiume poliuretaniche. Il lavoro di tesi da me svolto ha riguardato la messa a punto di una nuova metodologia sintetica per la preparazione del catalizzatore di Shvo e lo studio della sua attività catalitica nella riduzione di HMF a BHMF. Il comportamento del catalizzatore è stato monitorato studiando la resa in BHMF in funzione di tutti i parametri di reazione: temperatura, pressione di H2, solvente, rapporto molare substrato/catalizzatore, concentrazione, tempo. Successivamente è stata valutata la possibilità di riciclare il catalizzatore recuperando il prodotto di estrazione con acqua, per precipitazione o eseguendo la reazione in miscela bifasica (toluene/H2O). The present work is a collaboration between the Department of Physics and Inorganic Chemistry (group of Prof. Valerio Zanotti - Mattia Vaccari, Dr. Rita Mazzoni) and the Department of Industrial Chemistry and Materials (Group of Prof. Angelo Vaccari - Dr. Thomas Pasini, Dr. Stefania Albonetti, Prof. Fabrizio Cavani), and it’s a project devoted to evaluate the activity and selectivity of the Shvo catalyst, in the selective hydrogenation of polar double bond of 5 -hydroxymethylfurfural (HMF) in homogeneous phase. The HMF is an organic compound easily obtained from biomass, which can be used as a building block for fine chemicals abd polymer production or additives for biofuels with a high calorific value. In particular, our attention turned to the production of 2.5-bishydroxymethylfuran (BHMF), an important building block for the production of polymers and polyurethane foams. This thesis has involved the development of a new synthetic methodology for the preparation of Shvo’s catalyst and the study of its catalytic activity in the reduction of HMF to BHMF. The behavior of the catalyst was monitored by studying the yield in BHMF as a function of all the reaction parameters: temperature, pressure of H2, solvent, substrate to catalyst molar ratio, concentration, time. Subsequently it was evaluated the possibility of recycling the catalyst recovering the product of extraction with water, by precipitation or performing the reaction in biphasic mixture (toluene/H2O).

Latexpartikel als Trägermaterialien in der durch Postmetallocene katalysierten Ethenpolymerisation

Die vorliegende Arbeit beschäftigte sich mit der Immobilisierung von Postmetallocenen auf einem organischen Trägermaterial für die heterogene Ethenpolymerisation. Dabei konnte gezeigt werden, dass sich mit nukleophilen Gruppen funktionalisierte Latexpartikel als Trägermaterial für empfindliche Bis(phenoxyimin)titankatalysatoren eignen. Durch geschickte Kombination aus Trägermaterial, Katalysator und Cokatalysator wurde ein Katalysatorsysteme erhalten, das ultrahochmolekulares Polyethylen mit einem Molekulargewicht Mw von bis zu 7.000.000 g/mol (GPC, PS-Standard) bei einer enger Molekulargewichtsverteilung von weniger als 3 erzeugt. Die erhaltenen Produktivitäten erreichten dabei die Anforderungen an industriellen Katalysatorsystemen. Erstaunlich war, dass nukleophile Gruppen auf den Latexpartikeln, die dafür bekannt sind, dass sie den Katalysator deaktivieren können, ein wichtige Schutzfunktion für den Titankatalysator bilden. So konnte gezeigt werden, dass Pyridingruppen auf der Oberfläche der Latexpartikel als Scavenger gegen Trimethylaluminium wirken, welches ansonsten den aktiven Titankomplex zersetzen würde. An ausgewählten Systemen auf der Basis unterschiedlicher Postmetallocene und einem Metallocen für die Ethenpolymerisation wurde durch Anwendung verschiedener Methoden das Polymerisationsverhalten der Latex-geträgerten Katalysatorsysteme untersucht. Eine Methode war die so genannte Videomikroskopie, bei der in Gasphase Ethen polymerisiert wird und so das Wachstumsverhalten der Katalysatorpartikel analysiert werden kann. In Kombination mit einer kinetischen Kontrolle des Monomerumsatzes in Suspensionspolymerisation, konnte gezeigt werden, dass die auf Latexpartikel geträgerten Katalysatorsysteme ein anderes Polymerisationsverhalten besitzen wie in der Literatur beschriebene Silica-geträgerte Katalysatoren. Dies kann auf die unterschiedliche Beschaffenheit des Trägermaterials zurückgeführt werden. Während das harte Silica-Material Monomer nur in den Poren aufnimmt und so rasch nach Beginn der Polymerisation in eine diffusionskontrollierte Polymerisation übergeht, quillt das organische Trägermaterial mit dem Monomeren an und kann den aktiven Katalysator damit bedienen bis weiteres Monomer von außen in das Katalysatorpartikel nach diffundiert ist. Durch die weiche Beschaffenheit der organischen Latexpartikel kann das entstehende Polymer das Katalysatorpartikel außerdem leicht auseinandertreiben werden. Die Polymerisation kann so im ganzen Katalysatorpartikel beginnen, während das Silica-Trägermaterial nur langsam von außen nach innen fragmentieren kann.

Entwicklung massenspektrometrischer Isotopenverdünnungstechniken für die zuverlässige Bestimmung von Elementspuren und Schwefelspezies in Erdölprodukten

Einer der Hauptschwerpunkte der Arbeit lag in der Entwicklung einer spezies-spezifischen und einer spezies-unspezifischen GC-ICP-Q-MSIVA von Schwefelspezies in Petroprodukten. Es wurden hierzu Indikatoren, ausgehend von elementarem 34S-angereichertem Schwefel, im Mikromaßstab synthetisiert. Für die spezies-spezifische GC-ICP-Q-MSIVA wurde die erstmalige Synthese von 34S-markiertem Thiophen, Dibenzothiophen und 4-Methyldibenzothiophen verwirklicht. Als Indikatorsynthese für die spezies-unspezifische GC-ICP-Q-MSIVA erfolgte die erstmalige Darstellung von 34S-angereichertem Dimethyldisulid. Mit Hilfe der synthetisierten Verbindungen wurden spezies-spezifische und spezies-unspezifische massenspektrometrische Isotopenverdünnungsanalysen von Schwefelspezies in Petroprodukten durchgeführt. Vor allen GC-ICP-Q-MSIVA-Analysen erfolgte eine umfangreiche Speziesidentifizierung durch Aufstockexperimente mit kommerziell erhältlichen Standards und mit einem mit der GC gekoppelten Elektronenstoß (EI)-MS. Beide ICP-Q-MS Methoden zeichnen sich durch sehr niedrige Nachweisgrenzen (7 ng S/g) aus, welche auch eine Anwendbarkeit auf tiefentschwefelte Kraftstoffe garantieren. Mit der spezies-unspezifischen GC-ICP-Q-MSIVA ist neben einer Speziesanalyse auch eine Gesamtschwefelanalyse durch Aufsummierung aller in der Probe vorhandenen Spezies möglich. Es wurde im Rahmen dieser Arbeit auch der Einfluss möglicher Empfindlichkeitsänderungen des ICP-Q-MS durch koeluierende Kohlenwasserstoffe überprüft, wobei diese erwartungsgemäß auf das Ergebnis der spezies-spezifischen und spezies-unspezifischen GC-ICP-Q-MSIVA keinerlei Einfluss haben. Der zweite Hauptschwerpunkt der Arbeit lag auf der Ausarbeitung routinefähiger, schneller und zuverlässiger Methoden zur Gesamtelementspurenanalytik von Schwefel und Schwermetallen in Erdölen und Petroprodukten. Für die Gesamtschwefelanalyse wurde eine MSIVA nach thermaler Verdampfung mit 34S-markierten Dibenzothiophen als Indikator entwickelt. Die neu entwickelte Methode erlaubt eine sehr schnelle Bestimmung des Gesamtschwefelgehalts, wobei die eigentliche Messung des Isotopenverhältnisses innerhalb von Sekunden nach der Injektion der Probe erfolgt. Weiterhin zeichnet sich die Methode durch Robustheit und eine niedrige Nachweisgrenze (40 ng S/g) aus. Für die Analyse von Schwermetallen wurden erstmals Möglichkeiten einer direkten MSIVA von Erdölproben ohne zeitraubenden, kontaminationsträchtigen Aufschluss bzw. die schwierige Erzeugung einer Mikroemulsion zwischen hydrophober Probe und wässrigem Indikator entwickelt. Um eine homogene Verteilung des Indikators in der hydrophoben Probe zu ermöglichen, musste ausgehend von den zur Verfügung stehenden wässrigen Indikatorlösungen, eine Überführung des Indikators in ein organisches Lösungsmittel erfolgen. Hierzu wurde der jeweilige Metallindikator unter Komplexierung aus wässrigen Metallindikatorlösungen extrahiert. Für die Analyse der mit diesen Indikatorlösungen in organischer Phase versetzten Proben wurden zwei alternative Methoden ausgearbeitet. Bei der mit der Laserablation (LA) kombinierten ICP-SF-MSIVA wird die isotopenverdünnte Probe aus einer eigens für diesen Zweck entwickelten Probenhalterung ablatiert und so dem ICP-SF-MS zugeführt wird. Bei zeitlich sich verändernden Intensitäten der gemessenen Isotope werden aber reproduzierbare und konstante Isotopenverhältnisse erhalten. Im Falle einer homogenen Verteilung der Metallspuren wurde eine hervorragende Übereinstimmung mit Vergleichsmethoden und einem Referenzmaterial festgestellt. Im Falle einer heterogenen partikulären Verteilung der Metallspuren, wie sie z.B. bei Eisenspuren in den Erdölen vorlag, ist die Anwendbarkeit der LA-ICP-SF-MSIVA aufgrund des kleinen Probenvolumens (20 µL) jedoch begrenzt. Als Alternative zur LA-ICP-SF-MSIVA wurde ein System unter Verwendung der Fließinjektion für die Zuführung der isotopenverdünnten Probe zum ICP-SF-MS ausgearbeitet. Die isotopenverdünnte Probe wird hierbei in einen Eluentenstrom von Toluol injiziert und mit Hilfe einer Total-Consumption-Zerstäuber/Sprühkammer-Einheit vollständig bei einer Flussrate von 10 µL/min in das Plasma eingebracht. Neben einer nochmaligen Verkürzung der Analysenzeit und Vereinfachung der Probenvorbereitung bietet diese Methode zusätzlich stark verbesserte Nachweisgrenzen (z.B. Ni 0,9 ng/g). Leider sind mit diesem Verfahren bis jetzt nur Ni und Mo zuverlässig bestimmbar. Das in dieser Arbeit ausgearbeitete Methodenpaket erlaubt erstmals eine breite Einführung der ICP-MSIVA als zuverlässige Methode in die Routineanalytik der Petroindustrie. Durch die bewiesene Zuverlässigkeit, den geringen Zeitaufwand und die Robustheit der Methoden steht ihrem routinemäßigen Einsatz, außer einer weitergehenderen Automatisierung einzelner Verfahrensteile, prinzipiell nichts entgegen.

Quantum gravity effects in rotating black hole spacetimes

The aim of this work is to explore, within the framework of the presumably asymptotically safe Quantum Einstein Gravity, quantum corrections to black hole spacetimes, in particular in the case of rotating black holes. We have analysed this problem by exploiting the scale dependent Newton s constant implied by the renormalization group equation for the effective average action, and introducing an appropriate "cutoff identification" which relates the renormalization scale to the geometry of the spacetime manifold. We used these two ingredients in order to "renormalization group improve" the classical Kerr metric that describes the spacetime generated by a rotating black hole. We have focused our investigation on four basic subjects of black hole physics. The main results related to these topics can be summarized as follows. Concerning the critical surfaces, i.e. horizons and static limit surfaces, the improvement leads to a smooth deformation of the classical critical surfaces. Their number remains unchanged. In relation to the Penrose process for energy extraction from black holes, we have found that there exists a non-trivial correlation between regions of negative energy states in the phase space of rotating test particles and configurations of critical surfaces of the black hole. As for the vacuum energy-momentum tensor and the energy conditions we have shown that no model with "normal" matter, in the sense of matter fulfilling the usual energy conditions, can simulate the quantum fluctuations described by the improved Kerr spacetime that we have derived. Finally, in the context of black hole thermodynamics, we have performed calculations of the mass and angular momentum of the improved Kerr black hole, applying the standard Komar integrals. The results reflect the antiscreening character of the quantum fluctuations of the gravitational field. Furthermore we calculated approximations to the entropy and the temperature of the improved Kerr black hole to leading order in the angular momentum. More generally we have proven that the temperature can no longer be proportional to the surface gravity if an entropy-like state function is to exist.

The challenges and the limitations in Life Cycle Impact Assessment for metal oxide nanoparticles, a case study on nano- TiO2

Life Cycle Assessment (LCA) is a chain-oriented tool to evaluate the environment performance of products focussing on the entire life cycle of these products: from the extraction of resources, via manufacturing and use, to the final processing of the disposed products. Through all these stages consumption of resources and pollutant releases to air, water, soil are identified and quantified in Life Cycle Inventory (LCI) analysis. Subsequently to the LCI phase follows the Life Cycle Impact Assessment (LCIA) phase; that has the purpose to convert resource consumptions and pollutant releases in environmental impacts. The LCIA aims to model and to evaluate environmental issues, called impact categories. Several reports emphasises the importance of LCA in the field of ENMs. The ENMs offer enormous potential for the development of new products and application. There are however unanswered questions about the impacts of ENMs on human health and the environment. In the last decade the increasing production, use and consumption of nanoproducts, with a consequent release into the environment, has accentuated the obligation to ensure that potential risks are adequately understood to protect both human health and environment. Due to its holistic and comprehensive assessment, LCA is an essential tool evaluate, understand and manage the environmental and health effects of nanotechnology. The evaluation of health and environmental impacts of nanotechnologies, throughout the whole of their life-cycle by using LCA methodology. This is due to the lack of knowledge in relation to risk assessment. In fact, to date, the knowledge on human and environmental exposure to nanomaterials, such ENPs is limited. This bottleneck is reflected into LCA where characterisation models and consequently characterisation factors for ENPs are missed. The PhD project aims to assess limitations and challenges of the freshwater aquatic ecotoxicity potential evaluation in LCIA phase for ENPs and in particular nanoparticles as n-TiO2.

Pre-clinical and clinical development of leukemia stem cell inhibitors in acute leukemias

In Leukemias, recent developments have demonstrated that the Hedgehog pathway plays a key-role in the peculiar ability of self renewal of leukemia stem cells. The aim of this research activity was to investigate, through a first in man, Phase I, open label, clinical trial, the role and the impact, mainly in terms of safety profile, adverse events and pharmacokinetics, of a Sonic Hedgehog inhibitor compound on a population of heavely pretreated patients affected by AML, CML, MF, or MDS, resistant or refractory to standard chemotherapy. Thirty-five patients have been enrolled. The drug was administered orally, in 28 days cycles, without rest periods. The compound showed a good safety profile. The half life was of 17-35 hours, justifying the daily administration. Significant signs of activity, in terms of reduction of bone marrow blast cell amount were seen in most of the patients enrolled. Interestingly, correlative biological studies demonstrated that, comparing the gene expression profyiling signature of separated CD34+ cells before and after one cycle of treatment, the most variably expressed genes were involved in the Hh pathway. Moreover, we observed that many genes involved in MDR (multidrug resistance)were significantly down regulated after treatment. These data might lead to future clinical trials based on combinatory approaches, including, for instance, Hh inhibitors and conventional chemotherapy.