Comparison of the larynx-associated lymphoid tissue in the false vocal cords and subglottis in pediatric autopsies

The aim this work was to compare the distribution of cellular phenotypes of the LF in the FVC to the ones in the subglottic region in pediatric autopsy, relating this distribution to age and different causes of death. We analyzed 60 larynges of newborns and children autopsied in the period from 1993 to 2003. The fragments were prepared in order to perform histochemical and immunohistochemical techniques. The morphological analysis showed cases that presented LF only in FVC (35%), LF only in the subglottic region (20%), lack of LF in FVC (30%) and lymphoid aggregates, which did not characterize an LF (15%). The cases of LF in the subglottic region were significantly younger compared to the ones that presented LF in the FVC (p = 0.017). The LF in the subglottic region was bigger than the LF in the FVC (p = 0.020). There was no significant difference between the cause of death and cellular phenotype for both FVC and the subglottic region. In conclusion, the cells that make up the LF in the FVC in newborns and children younger than one year have functional characteristics similar to LF cells in the subglottic region, suggesting that there are similarities with LALT. (c) 2012 Elsevier GmbH. All rights reserved.

Records of sedimentary dynamics in the continental shelf and upper slope between Aveiro-Espinho (N Portugal)

The sedimentary unconsolidated cover of the Aveiro-Espinho continental shelf and upper slope (NW Portugal) records a complex interplay of processes including wave energy and currents, fluvial input, sediment transport alongshore and cross-shelf, geological and oceanographic processes and sediment sources and sinks. In order to study this record, a set of surface sediment samples was studied. Sediment grain size and composition, as well as the mineralogical composition (by XRD) of the fine (<63 mu m) and clay (<2 mu m) fractions and benthic microfaunal (foraminifera) data were analysed. Cluster analysis applied to the sedimentological data (grain size, sediment composition and mineralogy) allowed the establishment of three main zones corresponding to the: inner-, mid- and outer-shelf/upper slope. On the inner-shelf, the sedimentary coverture is composed of siliciclastic fine to very fine sand, essentially comprising modern (immature) terrigenous particles. The sediment grain size, as well as mineralogical and microfaunal composition, denote the high energetic conditions of this sector in which the alongshore transport of sand is predominantly southward and occurs mostly during the spring-summer oceanographic regime, when the main river providing sediments to this area, the River Douro, undergoes periods of drought. This effect may emphasize the erosive character of this coastal sector at present, since the Ria de Aveiro provides the shelf with few sediments. On the mid-shelf, an alongshore siliciclastic band of coarse sand and gravel can be found between the 40 m and 60 m isobaths. This gravelly deposit includes relic sediments deposited during lower sea-level stands. This structure stays on the surface due to the high bottom energy, which promotes the remobilization of the fine-grained sediments, and/or events of sediments bypassing. Benthic foraminifera density and "Benthic Foraminifera High Productivity" (BFHP) proxy values are in general low, which is consistent with the overall small supply of organic matter to the oceanic bottom in the inner- and mid-shelf. However, the Ria de Aveiro outflow, which delivers organic matter to the shelf, leaves its imprint mainly on the mid-shelf, identifiable by the increase in foraminifera density and BFHP values in front of the lagoon mouth. The higher values of BFHP along the 100 m isobath trace the present position of an oceanic thermal front whose situation may have changed in the last 3/5 ka BP. This zone marks a clear difference in the density, diversity and composition of benthic foraminifera assemblages. Here, in addition, sediment composition changes significantly, giving rise to carbonate-rich fine to medium sand in the deeper sector. The low bottom energy and the small sedimentation rate of the outer-shelf contributed to the preservation of a discontinuous carbonate-rich gravel band, between the 100 m and 140 m isobaths, also related to paleo-littorals, following the transgression that has occurred since the Last Glacial Maximum. The winter oceanographic regime favours the transport of fine grained sediments to the outer-shelf and upper slope. The inner- and mid-shelf, however, have low amounts of this kind of sediment and the Cretacic carbonated complexes Pontal da Galega and Pontal da Cartola, rocky outcrops located at the mid- and outer-shelf, act as morphological barriers to the cross-shelf transport of sediments. Thus a reduced sedimentation rate occurs in these deeper sectors, as indicated by the lower abundance of detrital minerals, which is compensated for the high sedimentary content of biogenic carbonates. The relatively high BFHP and Shannon Index values indicate water column stratification, high supply of organic matter and environmental stability, which provide favourable conditions for a diversified benthic fauna to flourish. These conditions also encourage authigenic chemical changes, favourable to glauconite formation, as well as illite and kaolinite degradation. Benthic foraminifera and clay mineral assemblages also reveal the effect of the internal waves pushing upward, and downslope losses of the sediments on the outer-shelf and upper slope.

Micro-structural and compositional variations of hydrothermal epidote-group minerals from a peralkaline granite, Corupá Pluton, Graciosa Province, South Brazil, and their petrological implications

Epidote-group minerals, together with albite, quartz, fluorite, Al-poor and Fe-rich phyllosilicates, zircon, and minor oxides and sulphides, are typical hydrothermal phases in peralkaline alkali-feldspar granites from the Corupá Pluton, Graciosa Province, South Brazil. The epidote-group minerals occur as single crystals and as aggregates filling in rock interstices and miarolitic cavities. They display complex recurrent zoning patterns with an internal zone of ferriallanite-(Ce), followed by allanite-(Ce), then epidote-ferriepidote, and an external zone with allanite-(Ce), with sharp limits, as shown in BSE and X-ray images. REE patterns show decreasing fractionation degrees of LREE over HREE from ferriallanite to epidote. The most external allanite is enriched in MREE. LA-ICP-MS data indicate that ferriallanite is enriched (>10-fold) in Ti, Sr and Ga, and depleted in Mg, Rb, Th and Zr relative to the host granite. Allanite has lower Ga and Mn and higher Zr, Nb and U contents as compared to ferriallanite, while epidote is enriched in Sr, U and depleted in Pb, Zr, Hf, Ti and Ga. The formation of these minerals is related to the variable concentrations of HFSE, Ca, Al, Fe and F in fluids remaining from magmatic crystallization, in an oxidizing environment, close to the HM buffer. L-MREE were in part released by the alteration of chevkinite, their main primary repository in the host rocks.

The influence of both urea and 2,2,2

The influence of external agents on proteins function and structure is essential to elucidate the unfolding pathways and self-assemble properties. The knowledge of the protein amyloid fibril formation process is important due to the fields that this subjected is related, in particular for the neurodegenerative disorders. In the present work we studied the influence of both urea and 2,2,2-Trifluoroethanol (TFE) and temperature on the structure and proteinprotein interactions of Bovine Serum Albumin (BSA), by means of UV-Vis spectroscopy, static fluorescence and small angle X-ray scattering technique. The experiments were performed in samples composed by 10 and 3 mg/ml of BSA at pH 5.8, near the protein pI. First, Thioflavin-T fluorescence measurements indicated that urea, in the absence of TFE, was able to increase the amyloid fibril formation of BSA at 45oC and increasing the urea concentration the rate of amyloid fibril formation also increases. Concerning the presence of TFE, SAXS data suggest that BSA tridimensional structure is not altered by the presence of TFE 5% and 10% v/v in all studied protein concentrations. Interestingly, the presence of TFE on the urea-containing BSA also increases the rate of amyloid fibril formation, as compared to the TFE-free system, indicating that TFE can catalyze the amyloid-fibril formation. The presence of TFE 20% v/v, however, induces the formation of aggregates, but at this time we were not able to infer if such aggregates are amyloidlike or amorphous. Taking together, the results give support to infer that BSA can for fibrils in the presence of urea at 45oC and TFE can act as a stabilizer or as a denaturant agent for BSA.

Morphology and surface sedimentation on the upper and middle continental slope off Uruguay

This paper presents for the first time a morphological and surface sediment characterization of the Uruguayan outer continental shelf and slope. The study is based on a high-resolution coverage using hydrographical, geomorphological and sedimentological sampling and several textural and productivity proxies. Along slope terraces and an important canyon system characterizes continental slope morphology, indicating that across- and down-slope sedimentary processes control large-scale sedimentation. Terraces represent the prolongation of the Argentinean Contouritic Depositional System that vanishes in the study area, presumably as a result of the dynamic of the Brazil-Malvinas confluence. Canyons incised in the upper slope are likely related to low-stand sea level conditions. At the outer shelf and shallow upper slope (170-250 m depth), off-shelf sand transport is inferred from the distribution of relict sand and reworked biogenic gravel. In the upper continental slope, the northern region is characterized by an erosive environment controlled by a steep slope and the southward flowing Brazil current. In the south, a depositional environment is enhanced by the presence of a gentler slope and seaward incised canyons and is mainly controlled by hemipelagic processes associated with nutrient-rich Sub-Antarctic Waters (SAW), by its confluence with South Atlantic Central Waters (SACW) and by the Rio de la Plata’s (RdlP) influence. Additionally, within the upper slope, the occurrence of igneous-metamorphic cobbles and pebbles in canyon and mound lag deposits suggests the influence of glacial fluvial discharge and/or iceberg transport processes. In the middle slope, sedimentation is controlled by thermohaline-induced deep-water bottom currents. The decreasing influence of the erosive Antarctic Intermediate Water (AAIW) is evident in a northward diminution in grain size. The variety of transport and sedimentary processes identified reflect the control of the Brazil-Malvinas confluence zone and the Rio de la Plata’s discharge.

Autophagic programmed cell death in the suspensor and endosperm of Vicia faba : An ultrastructural study

Programmed cell death (PCD) is a widely spread phenomenon among multi-cellular organisms. Without the deletion of cells no longer needed, the organism will not be able to develop in a predicted way. It is now belived that all cells have the capacity to self-destruct and that the survival of the cells is depending on the repression of this suicidal programme. PCD has turned out to show similarities in many different species and there are strong indications that the mechanisms running the programme might, at least in some parts, be evolutionarily conserced. PCD is a generic term for different programmes of cell destruction, such as apoptosis and autophagic PCD. An important tool to determine if a cell is undergoing PCD is the transmitting electron microscope. The aims of my study were to find out if, and in what way, the suspensor and endosperm in Vicia faba (Broad bean), which are short-lived structures, undergoes PCD. The endosperm degradation preceed the suspensor cell death and they differ to some extent ultrastructurally. The cell death occurs in both tissues about 13-14 days after pollination when the embryo proper is mature enough to support itself. It was found that both tissues are committed to autophagic PCD, a cell death characteristic of conspicuous formations of autophagic vacuoles. It was shown by histochemical staining that acid phosphatases are accumulated in these vacuoles but are also present in the cytoplasm. These vacuoles are similar to autophagic vacuoles formed in rat liver cells, indicating that autophagy is a widely spread phenomenon. DNA fragmentation is the first visible sign of PCD in both tissues and it is demonstrated by a labelling technique (TUNEL). In the endosperm nuclei the heterochromatin subsequently appears in the form of a network, while in the suspensor it is more conspicuous, with heterochromatin that forms large electron dense aggregates located close to the nuclear envelope. In the suspensor, the plastids develop into chromoplasts with lycopene crystals at the same time or shortly after DNA fragmentation. This is probably due to the fact that the suspensor plastids function as hormone producing organelles and support the embryo proper with indispensable growth factors. Later the embryo will be able to produce its own growth factors and the synthesis of these, in particular gibberelines, might be suppressed in the suspensor. The precursors can then be used for synthesis of lycopene instead. Both the suspensor and endosperm are going through autophagic PCD, but the process differs in some respects. This is probably due the the different function of the two tissues, and that the signals that trigger the process presumably are different. The embryo proper is probably the source of the death signal affecting the suspensor. The endosperm, which has a different origin and function, might be controlling the death signal within its own cell. The death might in this case be related to the age of the cell.

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.

Mixing properties of ester-containing solutions: a study on the ternary (methyl alkanoate (pentanoate and methanoate)+methanol and the corresponding binaries. New contributions to the (ester+ester) interactions

[EN]This work studies the volumetric (VE m) and energetic (HE m) properties resulting from the mixing processes of binary systems and the corresponding ternary of two methyl esters (methanoate and pentanoate) with methanol. The three binaries produce net endothermic mixing effects, with important energetic interactions, with maximum values of HE m ffi 400 J mol 1 , for the (ester + ester) system. This produces expansive effects VE m > 0, but the binaries of the (methyl esters + methanol) give rise to contractions VE m < 0, due to the formation of molecular aggregates.

Microfluidic device and interfacial transport: application to biomolecules and nanostructures

The aim of my dissertation is to provide new knowledge and applications of microfluidics in a variety of problems, from materials science, devices, and biomedicine, where the control on the fluid dynamics and the local concentration of the solutions containing the relevant molecules (either materials, precursors, or biomolecules) is crucial. The control of interfacial phenomena occurring in solutions at dierent length scales is compelling in nanotechnology for devising new sensors, molecular electronics devices, memories. Microfluidic devices were fabricated and integrated with organic electronics devices. The transduction involves the species in the solution which infills the transistor channel and confined by the microfluidic device. This device measures what happens on the surface, at few nanometers from the semiconductor channel. Soft-lithography was adopted to fabricate platinum electrodes, starting from platinum carbonyl precursor. I proposed a simple method to assemble these nanostructures in periodic arrays of microstripes, and form conductive electrodes with characteristic dimension of 600 nm. The conductivity of these sub-microwires is compared with the values reported in literature and bulk platinum. The process is suitable for fabricating thin conductive patterns for electronic devices or electrochemical cells, where the periodicity of the conductive pattern is comparable with the diusion length of the molecules in solution. The ordering induced among artificial nanostructures is of particular interest in science. I show that large building blocks, like carbon nanotubes or core-shell nanoparticles, can be ordered and self-organised on a surface in patterns due to capillary forces. The eective probability of inducing order with microfluidic flow is modeled with finite element calculation on the real geometry of the microcapillaries, in soft-lithographic process. The oligomerization of A40 peptide in microconfined environment represents a new investigation of the extensively studied peptide aggregation. The added value of the approach I devised is the precise control on the local concentration of peptides together with the possibility to mimick cellular crowding. Four populations of oligomers where distinguished, with diameters ranging from 15 to 200 nm. These aggregates could not be addresses separately in fluorescence. The statistical analysis on the atomic force microscopy images together with a model of growth reveal new insights on the kinetics of amyloidogenesis as well as allows me to identify the minimum stable nucleus size. This is an important result owing to its implications in the understanding and early diagnosis and therapy of the Alzheimer’s disease

Study of the building and decorative materials and techniques used in Hungarian Roman sites

The aims of this research were: - To identify the characteristics, properties and provenance of the building and decorative material found in three Hungarian Roman sites: Nagyharsány, Nemesvámos-Balácapuszta and Aquincum - To provide a database of information on the different sites - To have an overview of main conservation strategies applied in Hungary. Geological studies, macroscopical and microscopical observations, XRD investigations, physical and chemical analyses allowed us to define the characteristics and properties of the different kinds of collected materials. Building stones sampled from Nagyharsány site showed two different kinds of massive limestone belonging to the areas surrounding the villa. Also Building stones sampled from Nemesvámos-Balácapuszta Roman villa proved to be compatible with limestone belonging to local sources. Mural painting fragments show that all samples are units composed of multilayered structures. Mosaic tesserae can be classified as following: -Pale yellow , blackish and pink tesserae are comparable with local limestone; -White tessera, composed of marble, was probably imported from distant regions of the Empire, as the usual practice of Romans. Mortars present different characteristics according to the age, the site and the functions: -Building mortars are generally lime based, white or pale yellow in colour, present a high percentage of aggregates represented by fine sand; -Supporting mortars from both mosaics and mural paintings are reddish or pinkish in colour, due to the presence of high percentage of brick dust and tiles fragments, and present a higher content of MgO. Although the condition of the sites, there is an insignificant content of soluble salts. Database The whole study has allowed us to provide work sheets for each samples, including all characteristics and properties. Furthermore, all sites included in the frame of the research have been described and illustrated on the base of their floor plans, material and construction methodologies. It can be concluded that: 1. In Nagyharsány Archaeological site, it is possible to define a sequence of different construction phases on the base of the study of building material and mortars. The results are comparable with the chronology of the site provided by the archaeologists 2. The material used for construction was of local origin while the more precious ones, used for decorative elements, were probably imported from long distance 3. Construction techniques in Hungary mainly refer to the usual Roman knowledge and practice (Vitruvius); few differences have been found 4. The database will represent an archive for Archaeologists, Historians and Conservators dealing with Roman period in Hungary.

Kinematics of fibrous aggregates

This thesis reports on a research into the progressive development of fibrous aggregates, e.g. calcite, quartz and mica crystals in veins and strain fringes. The study is based on microstructural analysis of natural examples and on computer experiments. Investigation of fibrous looking elongate crystals in striped bedding-veins from the Orobic Alps, Italy indicate that these crystals do not track the opening trajectory of the veins but are oriented at an angle of up to 80° to the opening direction. Microstructural analysis of quartz, calcite and chlorite fibres in antitaxial strain fringes indicate that most strain fringes contain complex intergrowth of tracking (displacement-controlled) and non-tracking (face-controlled) fibres. To explain these growth features the computer program

Analytical challenges in the fields of Nanobioscience and Nanobiotecnology: integrated methods for separation and characterization

Nano(bio)science and nano(bio)technology play a growing and tremendous interest both on academic and industrial aspects. They are undergoing rapid developments on many fronts such as genomics, proteomics, system biology, and medical applications. However, the lack of characterization tools for nano(bio)systems is currently considered as a major limiting factor to the final establishment of nano(bio)technologies. Flow Field-Flow Fractionation (FlFFF) is a separation technique that is definitely emerging in the bioanalytical field, and the number of applications on nano(bio)analytes such as high molar-mass proteins and protein complexes, sub-cellular units, viruses, and functionalized nanoparticles is constantly increasing. This can be ascribed to the intrinsic advantages of FlFFF for the separation of nano(bio)analytes. FlFFF is ideally suited to separate particles over a broad size range (1 nm-1 μm) according to their hydrodynamic radius (rh). The fractionation is carried out in an empty channel by a flow stream of a mobile phase of any composition. For these reasons, fractionation is developed without surface interaction of the analyte with packing or gel media, and there is no stationary phase able to induce mechanical or shear stress on nanosized analytes, which are for these reasons kept in their native state. Characterization of nano(bio)analytes is made possible after fractionation by interfacing the FlFFF system with detection techniques for morphological, optical or mass characterization. For instance, FlFFF coupling with multi-angle light scattering (MALS) detection allows for absolute molecular weight and size determination, and mass spectrometry has made FlFFF enter the field of proteomics. Potentialities of FlFFF couplings with multi-detection systems are discussed in the first section of this dissertation. The second and the third sections are dedicated to new methods that have been developed for the analysis and characterization of different samples of interest in the fields of diagnostics, pharmaceutics, and nanomedicine. The second section focuses on biological samples such as protein complexes and protein aggregates. In particular it focuses on FlFFF methods developed to give new insights into: a) chemical composition and morphological features of blood serum lipoprotein classes, b) time-dependent aggregation pattern of the amyloid protein Aβ1-42, and c) aggregation state of antibody therapeutics in their formulation buffers. The third section is dedicated to the analysis and characterization of structured nanoparticles designed for nanomedicine applications. The discussed results indicate that FlFFF with on-line MALS and fluorescence detection (FD) may become the unparallel methodology for the analysis and characterization of new, structured, fluorescent nanomaterials.

Complexes of polyelectrolytes with defined charge distance and different dendrimer counterions

Complexes of polyelectrolytes with defined charge distance and different dendrimer counterions Magdalena Chelmecka Max Planck Institute for Polymer Research; Ackermannweg 10; D-55128 Mainz ; Tel.: (+49) 06131- 379 – 226 A study of complexes in solution is of interest to investigate whether the formation of well-defined assemblies like in classical surfactant systems is possible. Aim of this thesis is to investigate the electrostatic self-assembly of linear polycations of varying charge distance with “large” counterions of varying architecture. We especially investigate the morphology of objects formed, but also their stability under salt free condition and after low molecular mass salt addition. As polycations, Poly(dialkylimino)-alkylene salts (Ionenes) I65MeBr and I25MeBr were chosen. Ionenes are synthesized via Menschutkin reaction and characterized by standard methods. Counterions are Polyamidoamine (PAMAM) dendrimers of generations G2.5, G5.5, G7.5 with -COONa surface groups and shape-persistent, Polyphenylene dendrimers of generation G1 with surface -COOH groups. A complex interplay of interactions is expected to direct the self assembly via electrostatic interaction, geometric factors, hydrophobic interaction or hydrogen bonds. Methods used for the investigation of complexes are: UV-spectroscopy, pH-metric techniques, dynamic and static light scattering, small angle neutron scattering,  potential measurements and potentiometric titration. Under certain conditions, (i.e. charge ratio of compounds, charge density of ionene and dendrimer also concentration of sample) polyelectrolyte systems composed of ionenes and dendrimers build complexes in solution. System compounds are typical polyelectrolytes, but structures which they build behave not usual for typical polyelectrolytes. In a one diffusion mode regime aggregates of about 100 nm hydrodynamic radius have been found. Such aggregates are core-shell or anisotropic core shell structures in the case of ionenes/PAMAM dendrimers complexes. These complexes are stable even at high ionic strength. In case of ionenes with poly(phenylene) dendrimers, hard sphere-like objects or spherical objects with hairy-like surface have been found in a one diffusion mode regime. Their stability at high ionic strength is lower. For the ionenes/poly(phenylene) dendrimers systems one transition point has been found from one to two diffusion processes, towards increasing ionene concentration, i.e. for the samples with fixed dendrimer concentration towards increasing ionic strength. For the diffusion profile of ionene/PAMAM dendrimers in most cases two transition regimes are observed. One at very low ionene concentration, the second one at high ionene concentrations, which again means for the samples with fixed dendrimer concentration, also at higher ionic strength. Both two mode regimes are separated by the one mode regime. As was confirmed experimentally, the one diffusion mode regime is caused by the motion of well defined assemblies. The two diffusion mode regimes are caused by the movement of different sized species in solution, large aggregates and middle-size aggregates (oligoaggregates). The location and also the number of transition points in the diffusion profiles is dependent on the ionene to dendrimer charge ratio, charge density of the compounds and concentration. No influence of the molecular mass of the ionene has been found. The aggregates are found to be charged on the surface, however this surface charge does not significantly influence the diffusion properties of the system.

Liquid crystalline oligofluorenes and their derivatives

Homo-oligofluorenes (OFn), polyfluorenes (PF2/6) and oligofluorenes with one fluorenenone group in the center (OFnK) were synthesized. They were used as model compounds to understand of the structure-property relationships of polyfluorenes and the origin of the green emission in the photoluminescence (after photooxidation of the PFs) and the electroluminescence (EL) spectra. The electronic, electrochemical properties, thermal behavior, supramolecular self-assembly, and photophysical properties of OFn, PF2/6 and OFnK were investigated. Oligofluorenes with 2-ethylhexyl side chain (OF2-OF7) from the dimer up to the heptamer were prepared by a series of stepwise transition metal mediated Suzuki and Yamamoto coupling reactions. Polyfluorene was synthesized by Yamamoto coupling of 2,7-dibromo-9,9-bis(2-ethylhexyl)fluorene. Oligofluorenes with one fluorenone group in the center (OF3K, OF5K, OF7K) were prepared by Suzuki coupling between the monoboronic fluorenyl monomer, dimer, trimer and 2, 7-dibromofluorenone. The electrochemical and electronic properties of homo-oligofluorenes (OFn) were systematically studied by several combined techniques such as cyclic voltammetry, differential pulse voltammetry, UV-vis absorption spectroscopy, steady and time-resolved fluorescence spectroscopy. It was found that the oligofluorenes behave like classical conjugated oligomers, i.e., with the increase of the chain-length, the corresponding oxidation potential, the absorption and emission maximum, ionization potential, electron affinity, band gap and the photoluminescence lifetime displayed a very good linear relation with the reciprocal number of the fluorene units (1/n). The extrapolation of these linear relations to infinite chain length predicted the electrochemical and electronic properties of the corresponding polyfluorenes. The thermal behavior, single-crystal structure and supramolecular packing, alignment properties, and molecular dynamics of the homo-oligofluorenes (OFn) up to the polymer were studied using techniques such as TGA, DSC, WAXS, POM and DS. The OFn from tetramer to heptamer show a smectic liquid crystalline phase with clearly defined isotropization temperature. The oligomers do show a glass transition which exhibits n-1 dependence and allows extrapolation to a hypothetical glass transition of the polymer at around 64 °C. A smectic packing and helix-like conformation for the oligofluorenes from tetramer to heptamer was supported by WAXS experiments, simulation, and single-crystal structure of some oligofluorene derivatives. Oligofluorenes were aligned more easily than the corresponding polymer, and the alignability increased with the molecular length from tetramer to heptamer. The molecular dynamics in a series of oligofluorenes up to the polymer was studied using dielectric spectroscopy. The photophysical properties of OFn and PF2/6 were investigated by the steady-state spectra (UV-vis absorption and fluorescence spectra) and time-resolved fluorescence spectra both in solution and thin film. The time-resolved fluorescence spectra of the oligofluorenes were measured by streak camera and gate detection technique. The lifetime of the oligofluorenes decreased with the extension of the chain-length. No green emission was observed in CW, prompt and delayed fluorescence for oligofluorenes in m-THF and film at RT and 77K. Phosphorescence was observed for oligofluorenes in frozen dilute m-THF solution at 77K and its lifetime increased with length of oligofluorenes. A linear relation was obtained for triplet energy and singlet energy as a function of the reciprocal degree of polymerization, and the singlet-triplet energy gap (S1-T1) was found to decrease with the increase of degree of polymerization. Oligofluorenes with one fluorenone unit at the center were used as model compounds to understand the origin of the low-energy (“green”) emission band in the photoluminescence and electroluminescence spectra of polyfluorenes. Their electrochemical properties were investigated by CV, and the ionization potential (Ip) and electron affinity (Ea) were calculated from the onset of oxidation and reduction of OFnK. The photophysical properties of OFnK were studied in dilute solution and thin film by steady-state spectra and time-resolved fluorescence spectra. A strong green emission accompanied with a weak blue emission were obtained in solution and only green emission was observed on film. The strong green emission of OFnK suggested that rapid energy transfer takes place from higher energy sites (fluorene segments) to lower energy sites (fluorenone unit) prior to the radiative decay of the excited species. The fluorescence spectra of OFnK also showed solvatochromism. Monoexponential decay behaviour was observed by time-resolved fluorescence measurements. In addition, the site-selective excitation and concentration dependence of the fluorescence spectra were investigated. The ratio of green and blue emission band intensities increases with the increase of the concentration. The observed strong concentration dependence of the green emission band in solution suggests that increased interchain interactions among the fluorenone-containing oligofluorene chain enhanced the emission from the fluorenone defects at higher concentration. On the other hand, the mono-exponential decay behaviour and power dependence were not influenced significantly by the concentration. We have ruled out the possibility that the green emission band originates from aggregates or excimer formation. Energy transfer was further investigated using a model system of a polyfluorene doped by OFnK. Förster-type energy transfer took place from PF2/6 to OFnK, and the energy transfer efficiency increased with increasing of the concentration of OFnK. Efficient funneling of excitation energy from the high-energy fluorene segments to the low-energy fluorenone defects results from energy migration by hopping of excitations along a single polymer chain until they are trapped on the fluorenone defects on that chain or transferred onto neighbouring chains by Förster-type interchain energy transfer process. These results imply that the red-shifted emission in polyfluorenes can originate from (usually undesirable) keto groups at the bridging carbon atoms-especially if the samples have been subject to photo- or electro-oxidation or if fluorenone units are present due to an improper purification of the monomers prior to polymerization.

Impact of ocean acidification on phytoplankton growth and aggregation

Since the industrial revolution, the ocean has absorbed around one third of the anthropogenic CO2, which induced a profound alteration of the carbonate system commonly known as ocean acidification. Since the preindustrial times, the average ocean surface water pH has fallen by 0.1 units, from approximately 8.2 to 8.1 and a further decrease of 0.4 pH units is expected for the end of the century. Despite their microscopic size, marine diatoms are bio-geo-chemically a very important group, responsible for the export of massive amount of carbon to deep waters and sediments. The knowledge of the potential effects of ocean acidification on the phytoplankton growth and on biological pump is still at its infancy. This study wants to investigate the effect of ocean acidification on the growth of the diatom Skeletonema marinoi and on its aggregation, using a mechanistic approach. The experiment consisted of two treatments (Present and Future) representing different pCO2 conditions and two sequential experimental phases. During the cell growth phase a culture of S. marinoi was inoculated into transparent bags and the effect of ocean acidification was studied on various growth parameters, including DOC and TEP production. The aggregation phase consisted in the incubation of the cultures into rolling tanks where the sinking of particles through the water column was simulated and aggregation promoted. Since few studies investigated the effect of pH on the growth of S. marinoi and none used pH ranges that are compatible with the OA scenarios, there were no baselines. I have shown here, that OA does not affect the cell growth of S. marinoi, suggesting that the physiology of this species is robust in respect to the changes in the carbonate chemistry expected for the end of the century. Furthermore, according to my results, OA does not affect the aggregation of S. marinoi in a consistent manner, suggesting that this process has a high natural variability but is not influenced by OA in a predictable way. The effect of OA was tested over a variety of factors including the number of aggregates produced, their size and sinking velocity, the algal, bacterial and TEP content. Many of these variables showed significant treatment effects but none of these were consistent between the two experiments.