Electrodeposition of catalytic and magnetic gold nanoparticles on dendrimer-carbon nanotube layer-by-layer films

Magnetic and catalytic gold nanoparticles were electrodeposited through potential pulse on dendrimer-carbon nanotube layer-by-layer (LbL) films. A plasmon absorption band at about 550 nm revealed the presence of nanoscale gold in the film. The location of the Au nanoparticles in the film was clearly observed by selecting the magnetic force microscopy mode. To our knowledge, this is the first report on the electrochemical synthesis of magnetic Au nanoparticles. In addition to the magnetic properties, the Au nanoparticles also exhibited high catalytic activity towards ethanol and glycerol oxidation in alkaline medium.

Activation pattern of neutrophils from blood of elderly individuals with -related denture stomatitis

We have identified impaired neutrophils in elderly individuals which could be involved with -related denture stomatitis (DS), an oral infection predominantly caused by , affecting especially elderly individuals using dental prosthesis. However, specific mechanisms performed by neutrophil contributing to the susceptibility of the elderly to DS are not fully understood. This study evaluated activation features of blood neutrophils from elderly and young individuals with DS. Blood neutrophils cultured with . from elderly subjects secreted decreased levels of CXCL8. However, . challenged-neutrophils from DS patients produced high IL-4 and IL-10, and low GM-CSF levels, regardless of age. Additional elastase activity of neutrophils from both elderly groups was detected after incubation with . , but only neutrophils from elderly DS demonstrated high myeloperoxidase activity. Therefore, DS patients have affected neutrophils, and the advance of age intensifies these damages. In sumamry, individuals with -related denture stomatitis presented variation in the neutrophil phenotype and activation. Such alterations were more intense in neutrophils from infected elderly individuals.

Self-organized criticality in MHD driven plasma edge turbulence

We analyze long-range time correlations and self-similar characteristics of the electrostatic turbulence at the plasma edge and scrape-off layer in the Tokamak Chauffage Alfven Bresillien (TCABR), with low and high Magnetohydrodynamics (MHD) activity. We find evidence of self-organized criticality (SOC), mainly in the region near the tokamak limiter. Comparative analyses of data before and during the MHD activity reveals that during the high mHD activity the Hurst parameter decreases. Finally, we present a cellular automaton whose parameters are adjusted to simulate the analyzed turbulence SOC change with the MHD activity variation. (C) 2011 Published by Elsevier B.V.

Evaluation of immobilized lipases on poly-hydroxybutyrate beads to catalyze biodiesel synthesis

Five microbial lipase preparations from several sources were immobilized by hydrophobic adsorption on small or large poly-hydroxybutyrate (PHB) beads and the effect of the support particle size on the biocatalyst activity was assessed in the hydrolysis of olive oil, esterification of butyric acid with butanol and transesterification of babassu oil (Orbignya sp.) with ethanol. The catalytic activity of the immobilized lipases in both olive oil hydrolysis and biodiesel synthesis was influenced by the particle size of PHB and lipase source. In the esterification reaction such influence was not observed. Geobacillus thermocatenulatus lipase (BTL2) was considered to be inadequate to catalyze biodiesel synthesis, but displayed high esterification activity. Butyl butyrate synthesis catalyzed by BTL2 immobilized on small PHB beads gave the highest yield (approximate to 90 mmol L-1). In biodiesel synthesis, the catalytic activity of the immobilized lipases was significantly increased in comparison to the free lipases. Full conversion of babassu oil into ethyl esters was achieved at 72 h in the presence of Pseudozyma antarctica type B (CALB), Thermomyces lanuginosus lipase (Lipex (R) 100L) immobilized on either small or large PHB beads and Pseudomonas fluorescens (PFL) immobilized on large PHB beads. The latter preparation presented the highest productivity (40.9 mg of ethyl esters mg(-1) immobilized protein h(-1)). (C) 2012 Elsevier B.V. All rights reserved.

The impact of asymptomatic vertebral fractures on quality of life in older community-dwelling women: the Sao Paulo Ageing & Health Study

OBJECTIVES: The aim of this study was to investigate the impact of asymptomatic vertebral fractures on the quality of life in older women as part of the Sao Paulo Ageing & Health Study. METHODS: This study was a cross-sectional study with a random sample of 180 women 65 years of age or older with or without vertebral fractures. The Quality of Life Questionnaire of the European Foundation for Osteoporosis was administered to all subjects. Anthropometric data were obtained by physical examination, and the body mass index was calculated. Lateral thoracic and lumbar spine X-ray scans were obtained to identify asymptomatic vertebral fractures using a semi-quantitative method. RESULTS: Women with asymptomatic vertebral fractures had lower total scores [61.4(15.3) vs. 67.1(14.2), p = 0.03] and worse physical function domain scores [69.5(20.1) vs. 77.3(17.1), p = 0.02] for the Quality of Life Questionnaire of the European Foundation for Osteoporosis compared with women without fractures. The total score of this questionnaire was also worse in women classified as obese than in women classified as overweight or normal. High physical activity was related to a better total score for this questionnaire (p = 0.01). Likewise, lower physical function scores were observed in women with higher body mass index values (p < 0.05) and lower physical activity levels (p < 0.05). Generalized linear models with gamma distributions and logarithmic link functions, adjusted for age, showed that lower total scores and physical function domain scores for the Quality of Life Questionnaire of the European Foundation for Osteoporosis were related to a high body mass index, lower physical activity, and the presence of vertebral fractures (p < 0.05). CONCLUSION: Vertebral fractures are associated with decreased quality of life mainly physical functioning in older community-dwelling women regardless of age, body mass index, and physical activity. Therefore, the results highlight the importance of preventing and controlling asymptomatic vertebral fractures to reduce their impact on quality of life among older women.

Scorpiodinipora costulata (Canu & Bassler, 1929) (Bryozoa, Cheilostomata), a taxonomic and biogeographic dilemma: complex of cryptic species or human-mediated cosmopolitan colonizer?

Despite implausible cosmopolitanism, the species Scorpiodinipora costulata (Canu & Bassler, 1929) has been attributed with reservations to small encrusting colonies with similar morphological features whose known distribution is scattered in tropical and subtropical seas: Pacific Ocean (Philippines), Indian Ocean (Oman), Red Sea, SE Mediterranean, SE Atlantic (Ghana) and SW Atlantic (Brazil). This material raised questions about its generic assignment. The genus Scorpiodinipora Balavoine, 1959 is redescribed with Schizoporella costulata Canu & Bassler, 1929, from the Philippines as the type species, as Balavoine misidentified the specimens to define the genus as Cellepora bernardii Audouin, 1826. Moreover, SEM examination of the cotypes of S. costulata showed that Canu & Bassler confused two genera among them. A lectotype and paralectorype were thus chosen from Canu & Bassler's syntypes corresponding with the present morphotype. Hippodiplosia ottomuelleriana var. parva Marcus, 1938, from Brazil, which presents the same morphotype, is provisionally considered as the junior synonym of S. costulata. Considering the broad allopatric distribution of this morphotype across the oceans and the low capacity of dispersal of species with short-lived larvae, it is likely that this material includes several sibling species. However, the role of man-mediated dispersal is not excluded, at least in regions with high shipping activity, such as that comprising the Suez Canal.

The processing of polyelectrolyte-covered magnetite nanoparticles in the form of nanostructured thin films

Magnetic nanoparticles are promising for a variety of applications, such as biomedical devices, spin electronics, magnetic data storage media, to name a few. However, these goals may only be reached if stable and organized structures are fabricated. In this article, we report on a single-step synthetic route with the coprecipitation method, in which iron oxide magnetic nanoparticles (Fe3O4 NPs) were stabilized in aqueous media using the poly(diallyldimethylammonium chloride) (PDAC) polyelectrolyte. The Fe3O4 NPs had a diameter of ca. 5 nm, according to transmission electron microscopy (TEM) images, being arranged in an inverse spinel structure typical of magnetite. An investigation with infrared spectroscopy indicated that the mechanisms of stabilization in the polymer matrix were based on the interaction between quaternary amide groups from PDAC and the nanoparticle surface. The Fe3O4-PDAC NPs exhibited considerable magnetic susceptibility, with a monotonic increase in the magnetization with decreasing temperature. These Fe3O4-PDAC NPs were immobilized in layer-by-layer (LbL) films, being alternated with layers of poly(vinylsulfonic acid) (PVS). The LbL films were much rougher than typical films made with polyelectrolytes, and Fe3O4-PDAC NPs have been responsible for the high electrocatalytic activity toward H2O2 reduction, with an overpotential shift of 0.69 V. Overall, the stability, magnetic properties and film-forming ability indicate that the Fe3O4-PDAC NPs may be used for nanoelectronics and bioelectrochemical devices requiring reversible and magnetic redox materials.

Spectroscopic, morphological and electrochromic characterization of layer-by-layer hybrid films of polyaniline and hexaniobate nanoscrolls

The combination of semiconducting oxides and polyaniline in the nanoscale range may result in hybrid materials having enhanced properties, such as electrochromism and charge capacity. This paper reports the spectroscopic, morphological and electrochromic characterization of hybrid films made up of hexaniobate one-dimensional (1D) nanoscrolls and polyaniline prepared by the layer-by-layer assembly technique (LbL). Secondary electron imaging and backscattered electron imaging techniques performed using a scanning electron microscope showed that polyaniline is adsorbed on the hexaniobate nanoscrolls, which confirms the combination of the components in the nanoscale domain. UV-VIS-NIR electronic spectra of the LbL hybrid films showed the absorption tail in the NIR region, assigned to delocalized polarons of the polyaniline. Resonance Raman spectra in the 1000-1700 cm(-1) range indicated that hybrid films present a higher relative intensity of polaron bands at 1337 and 1508 cm(-1) than pristine polyaniline in the emeraldine salt form. These results suggest that hexaniobate nanoscrolls induce a secondary doping of polyaniline. The cyclic voltammetry (CV) data for the hybrid film showed a specific capacity of 870 C cm(-3). According to CV results, the synergistic effect on charge storage properties of the hybrid material is attributed to the enhanced electroactivity of the hexaniobate component in the LbL film. Spectroelectrochemical experiments showed that the electrochromic efficiencies at 420 nm are ca. -41 and 24 cm(2) C-1 as the potential changes from 0.8 to -0.9 V and from -0.9 to -1.8 V, respectively, whereas at 800 nm the efficiencies are ca. -55 and 8 cm(2) C-1 for the same potential ranges. The electrochromic efficiencies and multi-colour character of the LbL film of hexaniobate nanoscrolls and polyaniline indicate that this novel hybrid material is an interesting modified electrode for electrochromic devices.

The impact of asymptomatic vertebral fractures on quality of life in older community-dwelling women: the São Paulo Ageing & Health Study

OBJECTIVES: The aim of this study was to investigate the impact of asymptomatic vertebral fractures on the quality of life in older women as part of the Sao Paulo Ageing & Health Study. METHODS: This study was a cross-sectional study with a random sample of 180 women 65 years of age or older with or without vertebral fractures. The Quality of Life Questionnaire of the European Foundation for Osteoporosis was administered to all subjects. Anthropometric data were obtained by physical examination, and the body mass index was calculated. Lateral thoracic and lumbar spine X-ray scans were obtained to identify asymptomatic vertebral fractures using a semi-quantitative method. RESULTS: Women with asymptomatic vertebral fractures had lower total scores [61.4(15.3) vs. 67.1(14.2), p = 0.03] and worse physical function domain scores [69.5(20.1) vs. 77.3(17.1), p = 0.02] for the Quality of Life Questionnaire of the European Foundation for Osteoporosis compared with women without fractures. The total score of this questionnaire was also worse in women classified as obese than in women classified as overweight or normal. High physical activity was related to a better total score for this questionnaire (p = 0.01). Likewise, lower physical function scores were observed in women with higher body mass index values (p<0.05) and lower physical activity levels (p,0.05). Generalized linear models with gamma distributions and logarithmic link functions, adjusted for age, showed that lower total scores and physical function domain scores for the Quality of Life Questionnaire of the European Foundation for Osteoporosis were related to a high body mass index, lower physical activity, and the presence of vertebral fractures (p<0.05). CONCLUSION: Vertebral fractures are associated with decreased quality of life mainly physical functioning in older community-dwelling women regardless of age, body mass index, and physical activity. Therefore, the results highlight the importance of preventing and controlling asymptomatic vertebral fractures to reduce their impact on quality of life among older women.

Multiwavelenght study of cluster mergers and consequences for the radio emission properties of galaxy clusters

In the present thesis a thourough multiwavelength analysis of a number of galaxy clusters known to be experiencing a merger event is presented. The bulk of the thesis consists in the analysis of deep radio observations of six merging clusters, which host extended radio emission on the cluster scale. A composite optical and X–ray analysis is performed in order to obtain a detailed and comprehensive picture of the cluster dynamics and possibly derive hints about the properties of the ongoing merger, such as the involved mass ratio, geometry and time scale. The combination of the high quality radio, optical and X–ray data allows us to investigate the implications of the ongoing merger for the cluster radio properties, focusing on the phenomenon of cluster scale diffuse radio sources, known as radio halos and relics. A total number of six merging clusters was selected for the present study: A3562, A697, A209, A521, RXCJ 1314.4–2515 and RXCJ 2003.5–2323. All of them were known, or suspected, to possess extended radio emission on the cluster scale, in the form of a radio halo and/or a relic. High sensitivity radio observations were carried out for all clusters using the Giant Metrewave Radio Telescope (GMRT) at low frequency (i.e. ≤ 610 MHz), in order to test the presence of a diffuse radio source and/or analyse in detail the properties of the hosted extended radio emission. For three clusters, the GMRT information was combined with higher frequency data from Very Large Array (VLA) observations. A re–analysis of the optical and X–ray data available in the public archives was carried out for all sources. Propriety deep XMM–Newton and Chandra observations were used to investigate the merger dynamics in A3562. Thanks to our multiwavelength analysis, we were able to confirm the existence of a radio halo and/or a relic in all clusters, and to connect their properties and origin to the reconstructed merging scenario for most of the investigated cases. • The existence of a small size and low power radio halo in A3562 was successfully explained in the theoretical framework of the particle re–acceleration model for the origin of radio halos, which invokes the re–acceleration of pre–existing relativistic electrons in the intracluster medium by merger–driven turbulence. • A giant radio halo was found in the massive galaxy cluster A209, which has likely undergone a past major merger and is currently experiencing a new merging process in a direction roughly orthogonal to the old merger axis. A giant radio halo was also detected in A697, whose optical and X–ray properties may be suggestive of a strong merger event along the line of sight. Given the cluster mass and the kind of merger, the existence of a giant radio halo in both clusters is expected in the framework of the re–acceleration scenario. • A radio relic was detected at the outskirts of A521, a highly dynamically disturbed cluster which is accreting a number of small mass concentrations. A possible explanation for its origin requires the presence of a merger–driven shock front at the location of the source. The spectral properties of the relic may support such interpretation and require a Mach number M < ∼ 3 for the shock. • The galaxy cluster RXCJ 1314.4–2515 is exceptional and unique in hosting two peripheral relic sources, extending on the Mpc scale, and a central small size radio halo. The existence of these sources requires the presence of an ongoing energetic merger. Our combined optical and X–ray investigation suggests that a strong merging process between two or more massive subclumps may be ongoing in this cluster. Thanks to forthcoming optical and X–ray observations, we will reconstruct in detail the merger dynamics and derive its energetics, to be related to the energy necessary for the particle re–acceleration in this cluster. • Finally, RXCJ 2003.5–2323 was found to possess a giant radio halo. This source is among the largest, most powerful and most distant (z=0.317) halos imaged so far. Unlike other radio halos, it shows a very peculiar morphology with bright clumps and filaments of emission, whose origin might be related to the relatively high redshift of the hosting cluster. Although very little optical and X–ray information is available about the cluster dynamical stage, the results of our optical analysis suggest the presence of two massive substructures which may be interacting with the cluster. Forthcoming observations in the optical and X–ray bands will allow us to confirm the expected high merging activity in this cluster. Throughout the present thesis a cosmology with H0 = 70 km s−1 Mpc−1, m=0.3 and =0.7 is assumed.

Nuovi catalizzatori per l'idrogenazione, l'idrogenolisi/ring-opening di composti aromatici policiclici

The removal of aromatic hydrocarbons from diesel has received considerable attention after environmental regulations that require petroleum refiners to raise cetane number and to limit aromatics in diesel fuel in order to improve combustion efficiency and reduce particulate and NOx emissions. An alternative is blending with Fischer–Tropsch (FT) gas-to-liquid diesel fuel; however, this option may not be economically viable solution in case of extensive blend. Another alternative is to incorporate in the diesel pool a greater fraction of the so-called light cycle oil (LCO). Due to its high aromatics content and its low cetane number (typically between 20 and 30), the incorporation of LCO may have a negative impact on the quality of diesel. Current technologies for LCO improvement are based on hydrogenation to adjust both sulphur and cetane number but while an important fraction of the aromatics present in LCO can be saturated in a deep hydrogenation process, the cetane number may still be lower than the target values specified in diesel legislations, so further upgrading is needed. An interesting technology for improving the cetane number of diesels and maintaining meanwhile high diesel yields is achieved by combining a complete hydrogenation process with a selective ring opening (SRO) reaction of the naphthenic rings. The SRO can be defined as naphthene ring-opening to form compounds with high cetane number, but without any carbon losses. Controlling the interconversion of six- and five- membered rings via an acid-catalyzed ring-contraction step is also of great importance, since selective conversion of six-membered to five-membered naphthene rings greatly influences ring-opening rates and selectivity. High intrinsic activity may be enhanced by deposition of noble metals on acidic, high surface area supports, because it is possible to arrange close proximity of the metal and acid sites. Moreover, in large-pore supports, the diffusion resistance of liquid reactants into the pores is minimized. In addition to metal centres, the acid sites of support also plays role in aromatics hydrogenation. However, the functions of different kinds of acid sites (Brønsted vs. Lewis acidity), and their optimal concentrations and strengths, remain unclear. In the present study we investigated the upgrading of an aromatic-rich feedstock over different type of metal supported on mesoporous silica-alumina. The selective hydrogenolysis and ring opening of tetrahydronaphthalene (THN or tetralin) was carried out as representative of LCO fractions after deep hydrogenation process. In this regards the aim of this study is to evaluate both the effect of metals and that of the supports characterized by different acid distribution and strength, on conversion and selectivity. For this purpose a series of catalysts were prepared by impregnation. The catalysts were characterized and conversion tests of THN were performed in a lab-scale plant operating in the pressure range from 7.0-5.0 MPa and in the temperature range from 300 to 360°C.

Determination of bioactive and potentially toxic compounds in food: raw material analysis and shelf life evaluation

"Bioactive compounds" are extranutritional constituents that typically occur in small quantities in food. They are being intensively studied to evaluate their effects on health. Bioactive compounds include both water soluble compounds, such as phenolics, and lipidic substances such as n-3 fatty acids, tocopherols and sterols. Phenolic compounds, tocopherols and sterols are present in all plants and have been studied extensively in cereals, nuts and oil. n-3 fatty acids are present in fish and all around the vegetable kingdom. The aim of the present work was the determination of bioactive and potentially toxic compounds in cereal based foods and nuts. The first section of this study was focused on the determination of bioactive compounds in cereals. Because of that the different forms of phytosterols were investigated in hexaploid and tetraploid wheats. Hexaploid cultivars were the best source of esterified sterols (40.7% and 37.3% of total sterols for Triticum aestivum and Triticum spelta, respectively). Significant amounts of free sterols (65.5% and 60.7% of total sterols for Triticum durum and Triticum dicoccon, respectively) were found in the tetraploid cultivars. Then, free and bound phenolic compounds were identified in barley flours. HPLCESI/ MSD analysis in negative and positive ion mode established that barley free flavan-3- ols and proanthocyanidins were four dimers and four trimers having (epi)catechin and/or (epi)gallocatechin (C and/or GC) subunits. Hydroxycinnamic acids and their derivatives were the main bound phenols in barley flours. The results obtained demonstrated that barley flours were rich in phenolic compounds that showed high antioxidant activity. The study also examined the relationships between phenolic compounds and lipid oxidation of bakery. To this purpose, the investigated barley flours were used in the bakery production. The formulated oven products presented an interesting content of phenolic compounds, but they were not able to contain the lipid oxidation. Furthermore, the influence of conventional packaging on lipid oxidation of pasta was evaluated in n-3 enriched spaghetti and egg spaghetti. The results proved that conventional packaging was not appropriated to preserve pasta from lipid oxidation; in fact, pasta that was exposed to light showed a high content of potentially toxic compounds derived from lipid oxidation (such as peroxide, oxidized fatty acids and COPs). In the second section, the content of sterols, phenolic compounds, n-3 fatty acids and tocopherols in walnuts were reported. Rapid analytical techniques were used to analyze the lipid fraction and to characterize phenolic compounds in walnuts. Total lipid chromatogram was used for the simultaneous determination of the profile of sterols and tocopherols. Linoleic and linolenic acids were the most representative n-6 and n-3 essential dietary fatty acids present in these nuts. Walnuts contained substantial amounts of γ- and δ-tocopherol, which explained their antioxidant properties. Sitosterol, Δ5-avenasterol and campesterol were the major free sterols found. Capillary electrophoresis coupled to DAD and microTOF was utilized to determine phenolic content of walnut. A new compound in walnut ((2E,4E)- 8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 6-O-β-D-glucopiranosyl ester, [M−H]− 403.161m/z) with a structure similar to glansreginins was also identified. Phenolic compounds corresponded to 14–28% of total polar compounds quantified. Aglycone and glycosylated ellagic acid represented the principal components and account for 64–75% of total phenols in walnuts. However, the sum of glansreginins A, B and ((2E,4E)-8-hydroxy- 2,7-dimethyl-2,4-decadiene-1,10-dioic acid 6-O-β-D-glucopiranosyl ester was in the range of 72–86% of total quantified compounds.

A candidate gene approach to identify DNA markers associated with meat quality and production traits: investigation of several cathepsin genes in Italian heavy pigs.

The cathepsin enzymes represent an important family of lysosomal proteinases with a broad spectrum of functions in many, if not in all, tissues and cell types. In addition to their primary role during the normal protein turnover, they possess highly specific proteolytic activities, including antigen processing in the immune response and a direct role in the development of obesity and tumours. In pigs, the involvement of cathepsin enzymes in proteolytic processes have important effects during the conversion of muscle to meat, due to their influence on meat texture and sensory characteristics, mainly in seasoned products. Their contribution is fundamental in flavour development of dry-curing hams. However, several authors have demonstrated that high cathepsin activity, in particular of cathepsin B, is correlated to defects of these products, such as an excessive meat softness together with abnormal free tyrosine content, astringent or metallic aftertastes and formation of a white film on the cut surface. Thus, investigation of their genetic variability could be useful to identify DNA markers associated with these dry cured hams parameters, but also with meat quality, production and carcass traits in Italian heavy pigs. Unfortunately, no association has been found between cathepsin markers and meat quality traits so far, in particular with cathepsin B activity, suggesting that other genes, besides these, affect meat quality parameters. Nevertheless, significant associations were observed with several carcass and production traits in pigs. A recent study has demonstrated that different single nucleotide polymorphisms (SNPs) localized in cathepsin D (CTSD), F (CTSF), H and Z genes were highly associated with growth, fat deposition and production traits in an Italian Large White pig population. The aim of this thesis was to confirm some of these results in other pig populations and identify new cathepsin markers in order to evaluate their effects on cathepsin activity and other production traits. Furthermore, starting from the data obtained in previous studies on CTSD gene, we also analyzed the known polymorphism located in the insulin-like growth factor 2 gene (IGF2 intron3-g.3072G>A). This marker is considered the causative mutation for the quantitative trait loci (QTL) affecting muscle mass and fat deposition in pigs. Since IGF2 maps very close to CTSD on porcine chromosome (SSC) 2, we wanted to clarify if the effects of the CTSD marker were due to linkage disequilibrium with the IGF2 intron3-g.3072G>A mutation or not. In the first chapter, we reported the results from these two SSC2 gene markers. First of all, we evaluated the effects of the IGF2 intron3-g.3072G>A polymorphism in the Italian Large White breed, for which no previous studies have analysed this marker. Highly significant associations were identified with all estimated breeding values for production and carcass traits (P<0.00001), while no effects were observed for meat quality traits. Instead, the IGF2 intron3-g.3072G>A mutation did not show any associations with the analyzed traits in the Italian Duroc pigs, probably due to the low level of variability at this polymorphic site for this breed. In the same Duroc pig population, significant associations were obtained for the CTSD marker for all production and carcass traits (P < 0.001), after excluding possible confounding effects of the IGF2 mutation. The effects of the CTSD g.70G>A polymorphism were also confirmed in a group of Italian Large White pigs homozygous for the IGF2 intron3-g.3072G allele G (IGF2 intron3-g.3072GG) and by haplotype analysis between the markers of the two considered genes. Taken together, all these data indicated that the IGF2 intron3-g.3072G>A mutation is not the only polymorphism affecting fatness and muscle deposition in pigs. In the second chapter, we reported the analysis of two new SNPs identified in cathepsin L (CTSL) and cathepsin S (CTSS) genes and the association results with meat quality parameters (including cathepsin B activity) and several production traits in an Italian Large White pig population. Allele frequencies of these two markers were evaluated in 7 different pig breeds. Furthermore, we mapped using a radiation hybrid panel the CTSS gene on SSC4. Association studies with several production traits, carried out in 268 Italian Large White pigs, indicated positive effects of the CTSL polymorphism on average daily gain, weight of lean cuts and backfat thickness (P<0.05). The results for these latter traits were also confirmed using a selective genotype approach in other Italian Large White pigs (P<0.01). In the 268 pig group, the CTSS polymorphism was associated with feed:gain ratio and average daily gain (P<0.05). Instead, no association was observed between the analysed markers and meat quality parameters. Finally, we wanted to verify if the positive results obtained for the cathepsin L and S markers and for other previous identified SNPs (cathepsin F, cathepsin Z and their inhibitor cystatin B) were confirmed in the Italian Duroc pig breed (third chapter). We analysed them in two groups of Duroc pigs: the first group was made of 218 performance-tested pigs not selected by any phenotypic criteria, the second group was made of 100 Italian Duroc pigs extreme and divergent for visible intermuscular fat trait. In the first group, the CTSL polymorphism was associated with weight of lean cuts (P<0.05), while suggestive associations were obtained for average daily gain and backfat thickness (P<0.10). Allele frequencies of the CTSL gene marker also differed positively among the visible intermuscular extreme tails. Instead, no positive effects were observed for the other DNA markers on the analysed traits. In conclusion, in agreement with the present data and for the biological role of these enzymes, the porcine CTSD and CTSL markers: a) may have a direct effect in the biological mechanisms involved in determining fat and lean meat content in pigs, or b) these markers could be very close to the putative functional mutation(s) present in other genes. These findings have important practical applications, in particular the CTSD and CTSL mutations could be applied in a marker assisted selection (MAS) both in the Italian Large White and Italian Duroc breeds. Marker assisted selection could also increase in efficiency by adding information from the cathepsin S genotype, but only in the Italian Large White breed.

Selektive und duale COX-1-, COX-2-Inhibitoren aus der Reihe der Methanone

Zusammenfassung: Prostaglandine (PG) sind wichtige biologische Entzündungsmediatoren, die aus der Arachidonsäure (AA) durch das Enzym Cyclooxygenase (COX) entstehen. Trotz einiger unerwünschter Wirkungen, sind Cyclooxygenase-Hemmer Mittel der Wahl zur Unterdrückung entzündlicher Prozesse. Von der Cyclooxygenase existieren zwei Isoenzyme: COX-1 und COX-2. Eine selektive Hemmung der COX-2 bzw. eine duale Hemmung der COX-1 und COX-2 wird als erfolgversprechendes Prinzip zur Behandlung von entzündlichen Erkrankungen diskutiert.Ziel der Arbeit war die Synthese und in vitro Testung sowie die Erstellung von Struktur-Wirkungs-Beziehungen selektiver bzw. dualer Hemmstoffe der COX-1/-2. Zusätzlich wurden die Substanzen auf inhibitorische Aktivität gegenüber der 5- und 12-Lipoxygenase untersucht.Ausgehend von der Struktur selektiver Hemmstoffe der COX-2 bzw. von dualen COX-1/ COX-2-Inhibitoren sowie von marktüblichen nichtsteroidalen Antirheumatika (NSAR), wurde das Diarylmethanon-Element als Basis gewählt. An diesem Strukturelement wurden Modifikationen vorgenommen, um selektive Hemmstoffe der COX-2 bzw. duale COX-1/ COX-2-Hemmstoffe zu erhalten.Die synthetisierten Verbindungen lassen sich in [4-(Methylsulfanyl)phenyl]- und [4-(Methylsulfonyl)phenyl](aryl)methanone, N-(Aroylphenyl)sulfonamide und -amide sowie (Hydroxyphenyl)(2-thienyl)methanone unterteilen.In der Reihe der [4-(Methylsulfanyl)phenyl](aryl)methanone sind potente Hemmstoffe sowohl der COX-1 als auch der COX-2 erhalten worden. Im Gegensatz dazu zeigen die [4-(Methylsulfonyl)phenyl](aryl)methanone gegenüber COX-1 und COX-2 keine inhibitorische Aktivität. Mit dem 2-Thienylderivat wurde ein potenter, dualer Hemmstoff beider Cyclooxygenase-Isoenzyme identifiziert, dessen Wirkstärke (bezüglich der COX-2) auf den Austausch von Phenyl gegen 2-Thienyl zurückzuführen ist.Die N-(Aroylphenyl)sulfonamide und -amide bilden die umfangreichste Gruppe bei den durchgeführten Untersuchungen, wobei besonders die regioisomeren N-(2-Aroylphenyl)sulfonamide und -amide eingehender studiert wurden. Auf der Basis der (2-Aroylphenyl)sulfonamide läßt sich für die Hemmung der COX-1 eine Struktur-Wirkungs-Beziehungen formulieren, die anhand Hilfe geeigneter Verbindungen überprüft wurde. Die Untersuchungen wurden zum Teil auch auf die 3- und 4-Regioisomeren ausgedehnt, wobei sich die erhaltenen Struktur-Wirkungs-Beziehungen bestätigten. Die Arylsulfonamide inhibieren bevorzugt die COX-1. Auch (4-Aroylphenyl)sulfonamide wurden auf mögliche inhibitorische Aktivität untersucht. Die Einbindung des Amidstickstoffs in ein Indolin- bzw. Tetrahydrochinolin-Ringsystem oder des Sulfonamids in ein 1,3-Propansultam führte in jedem Falle zu wenig aktiven Verbindungen gegenüber der COX-1. N-(2-Aroylphenyl)amide zeigten in Übereinstimmung mit der Hypothese an der COX-1 eine gute inhibitorische Aktivität.Aus der Reihe der (Hydroxyphenyl)(2-thienyl)methanone wurden die freien Alkohole, die Methylether und verschiedene Ester dargestellt und auf COX-1-Aktivität untersucht. Acetate, aber auch Phenole sind die potentesten Inhibitoren der COX-1. Als günstigte Positionen für die 2-Thienylcarbonyl-Einheit am Hydroxyphenylrest erweist sich die ortho- bzw. para-Position.

Differenzielle Expression von Tubulin-Genen bei der Entwicklung von Blattzellen der Gerste (Hordeum vulgare L.)

Die Morphogenese einer Pflanzenzelle wird in großem Maße durch die Dynamik kortikaler Mikrotubuli (MT) bestimmt, die auf die Zellwandsynthese Einfluß nehmen. In dieser Arbeit wurden die Transkriptmengen der alpha-Tubulin-Isotypen und des gamma-Tubulin während der Entwicklung des Gerstenblattes analysiert, um Zusammenhänge zu bereits beschriebenen Umwandlungen im kortikalen MT-Cytoskelett der Mesophyllzellen aufzudecken. Erstmals konnte bei einer höheren Pflanze die Genexpression auf RNA-Ebene innerhalb einer Tubulin-Multigenfamilie im Verlauf der Blattentwicklung umfassend dargestellt werden.Es wurden blattspezifische cDNA-Bibliotheken erstellt und mittels RT-PCR homologe DNA-Gensonden für die Screeningprozesse der cDNA-Bibliotheken hergestellt. cDNA-Sequenzen von alpha-, beta-, und gamma-Tubulin konnten isoliert werden. Weitere, weniger abundante alpha-Tubulin-Sequenzen wurden während zusätzlicher Screeningrunden über PCR-Ausschluß häufig vertretener, bereits bekannter Isotypen isoliert.Die cDNA-Sequenzen von insgesamt fünf verschiedenen Isotypen des alpha-Tubulin konnten aufgeklärt werden, drei Isotypen wiesen bis zu fünf im nicht kodierenden 3´-Bereich verkürzte Varianten auf, die aber in ihrer Anzahl deutlich unterrepräsentiert waren. Die abgeleiteten Aminosäuresequenzen umfassten bei drei Isotypen 451 Aminosäuren (AS), zwei Isotypen waren im C-Terminus um eine bzw. um zwei AS kürzer. Die fünf alpha-Tubulin-Isotypen wiesen charakteristische Expressionsmuster auf, die in drei Klassen unterteilbar waren. Die Isotypen HVATUB1 und HVATUB5 (MT-Band-Isotypen) hatten den maximalen Gehalt in Blattbereichen, in denen auch hauptsächlich Mesophyllzellen mit kortikalen MT-Bänderungen vorkommen, wobei HVATUB5 den am schwächsten exprimierte Isotyp darstellte. HVATUB3 (Random-MT-Isotyp) zeigte die stärksten Expressionsraten. Die im Meristem und meristemnahen Bereichen bereits recht hohe Abundanz erreichte erst nach der Zellstreckungszone in einer Blattzone das Maximum, in dem hauptsächlich Mesophyllzellen mit zerstreut angeordneten MT anzutreffen sind. Die Isotypen HVATUB2 und HVATUB4 (MImax-Isotypen) waren in mitotisch aktiven, basalen Blattbereichen dominant.Die cDNA-Sequenz vom gamma-Tubulin der Gerste, HVGTUB, wurde ermittelt; die abgeleitete Aminosäuresequenz bestand aus 469 AS. Das Auftreten einer im nicht kodierenden 3´-Bereich kürzeren Variante konnte erstmals bei pflanzlichem gamma-Tubulin beschrieben werden. Southernblot-Analysen ließen darauf schließen, daß gamma-Tubulin nur als Einzelkopie im Genom der Gerste vorkommt. gamma-Tubulin wurde im mitosereichen Meristem der Blattbasis am stärksten exprimiert. Da die Abnahme der Transkriptmenge weitaus langsamer verlief als die Abnahme der Zellteilungsaktivität, ist anzunehmen, daß gamma-Tubulin neben der Erfüllung von mitose- und zellteilungsspezifischen Funktionen auch eine Rolle im Zusammenhang mit der Dynamik des kortikalen MT-Cytoskeletts spielt. Einen ersten Schritt zur Aufklärung der Genfamilie des beta-Tubulin bei Gerste stellt die Isolierung drei verschiedener cDNA-Sequenzen von beta-Tubulin dar.