Incidence of lower limb bone fractures in female individuals of Aracaju (Sergipe), Brazil

Studies indicate that the number of hip bone fractures caused by osteoporosis may rise from 1.66 to 6.26 million until 2050, worldwide. For this reason, implementation of preventive measures becomes a necessity. Female individuals are usually more affected due to a variety of factors including old age, early menopause, chronicle disease in the family history, calcium deficit, as well as the lack of physical exercise (sedentary individual). The aim of this study was to estimate the incidence of hip and lower limb fracture in female individuals’ resident in Aracaju city. From the period of January 2008-2009, around of 300 fracture cases were of lower limb analyzed from females. The incidence of femur fractures in women increased according to age group, 66.17 individuals per 10,000 inhabitants (over 60 years-old). These findings allow us to conclude that the incidence of hip and lower limb bone fractures among women over 60 years were more significant in the femur.

Exercise and caloric restriction alter the immune system of mice submitted to a high-fat diet

As the size of adipocytes increases during obesity, the establishment of resident immune cells in adipose tissue becomes an important source of proinflammatory mediators. Exercise and caloric restriction are two important, nonpharmacological tools against body mass increase. To date, their effects on the immune cells of adipose tissue in obese organisms, specifically when a high-fat diet is consumed, have been poorly investigated. Thus, after consuming a high-fat diet, mice were submitted to chronic swimming training or a 30% caloric restriction in order to investigate the effects of both interventions on resident immune cells in adipose tissue. These strategies were able to reduce body mass and resulted in changes in the number of resident immune cells in the adipose tissue and levels of cytokines/chemokines in serum. While exercise increased the number of NK cells in adipose tissue and serum levels of IL-6 and RANTES, caloric restriction increased the CD4+/CD8+ cell ratio and MCP-1 levels. Together, these data demonstrated that exercise and caloric restriction modulate resident immune cells in adipose tissues differently in spite of an equivalent body weight reduction. Additionally, the results also reinforce the idea that a combination of both strategies is better than either individually for combating obesity

Efecto del color del oponente en el comportamiento agresivo del Grapsus grapsus (Linnaeus, 1758) (Crustacea, Decapoda)

[ES] El comportamiento agresivo del cangrejo moro (Grapsus grapsus) ante un modelo de competidor potencial no varió en función del color corporal de este último. No obstante, se observaron cambios en dicho comportamiento en función del tamaño del individuo residente, aumentado la intensidad de los ataques cuanto menores eran las asimetrías en talla

Territorio, participación ciudadana y nivel de confianza en las instituciones oficiales

[ES] Canarias es una de las regiones españolas que registra mayores niveles de abstención en las diferentes consultas electorales. Este comportamiento puede explicarse por razones de diverso tipo, pero sin duda, la falta de confianza en los cargos públicos electos y en las instituciones es uno de los factores de mayor peso. Con este fin, en la presente comunicación se expone resumidamente una reciente experiencia realizada en Canarias por el Grupo de Investigación en Geografía Social y Económica de la ULPGC consistente en situar las diferentes respuestas que dio una muestra de ciudadanos residentes en las Islas a una encuesta de opinión confeccionada al efecto sobre el grado de confianza que le inspiran las instituciones oficiales. A su vez, los opinantes fueron desagregados por islas, comarcas y municipios, poniéndose en evidencia la enorme pluralidad y complejidad que es capaz de albergar una sociedad moderna como la canaria. 

An insight into the populations of blainville's and curvier's beaked whales off El Hierro (Canary Islands)

Trabajo realizado por: Reyes, C., Schiavi, A., Aguilar del Soto,

RNA Interference and cyclooxygenase-2 (COX-2) regulation in colon cancer cells

Despite new methods and combined strategies, conventional cancer chemotherapy still lacks specificity and induces drug resistance. Gene therapy can offer the potential to obtain the success in the clinical treatment of cancer and this can be achieved by replacing mutated tumour suppressor genes, inhibiting gene transcription, introducing new genes encoding for therapeutic products, or specifically silencing any given target gene. Concerning gene silencing, attention has recently shifted onto the RNA interference (RNAi) phenomenon. Gene silencing mediated by RNAi machinery is based on short RNA molecules, small interfering RNAs (siRNAs) and microRNAs (miRNAs), that are fully o partially homologous to the mRNA of the genes being silenced, respectively. On one hand, synthetic siRNAs appear as an important research tool to understand the function of a gene and the prospect of using siRNAs as potent and specific inhibitors of any target gene provides a new therapeutical approach for many untreatable diseases, particularly cancer. On the other hand, the discovery of the gene regulatory pathways mediated by miRNAs, offered to the research community new important perspectives for the comprehension of the physiological and, above all, the pathological mechanisms underlying the gene regulation. Indeed, changes in miRNAs expression have been identified in several types of neoplasia and it has also been proposed that the overexpression of genes in cancer cells may be due to the disruption of a control network in which relevant miRNA are implicated. For these reasons, I focused my research on a possible link between RNAi and the enzyme cyclooxygenase-2 (COX-2) in the field of colorectal cancer (CRC), since it has been established that the transition adenoma-adenocarcinoma and the progression of CRC depend on aberrant constitutive expression of COX-2 gene. In fact, overexpressed COX-2 is involved in the block of apoptosis, the stimulation of tumor-angiogenesis and promotes cell invasion, tumour growth and metastatization. On the basis of data reported in the literature, the first aim of my research was to develop an innovative and effective tool, based on the RNAi mechanism, able to silence strongly and specifically COX-2 expression in human colorectal cancer cell lines. In this study, I firstly show that an siRNA sequence directed against COX-2 mRNA (siCOX-2), potently downregulated COX-2 gene expression in human umbilical vein endothelial cells (HUVEC) and inhibited PMA-induced angiogenesis in vitro in a specific, non-toxic manner. Moreover, I found that the insertion of a specific cassette carrying anti-COX-2 shRNA sequence (shCOX-2, the precursor of siCOX-2 previously tested) into a viral vector (pSUPER.retro) greatly increased silencing potency in a colon cancer cell line (HT-29) without activating any interferon response. Phenotypically, COX-2 deficient HT-29 cells showed a significant impairment of their in vitro malignant behaviour. Thus, results reported here indicate an easy-to-use, powerful and high selective virus-based method to knockdown COX-2 gene in a stable and long-lasting manner, in colon cancer cells. Furthermore, they open up the possibility of an in vivo application of this anti-COX-2 retroviral vector, as therapeutic agent for human cancers overexpressing COX-2. In order to improve the tumour selectivity, pSUPER.retro vector was modified for the shCOX-2 expression cassette. The aim was to obtain a strong, specific transcription of shCOX-2 followed by COX-2 silencing mediated by siCOX-2 only in cancer cells. For this reason, H1 promoter in basic pSUPER.retro vector [pS(H1)] was substituted with the human Cox-2 promoter [pS(COX2)] and with a promoter containing repeated copies of the TCF binding element (TBE) [pS(TBE)]. These promoters were choosen because they are partculary activated in colon cancer cells. COX-2 was effectively silenced in HT-29 and HCA-7 colon cancer cells by using enhanced pS(COX2) and pS(TBE) vectors. In particular, an higher siCOX-2 production followed by a stronger inhibition of Cox-2 gene were achieved by using pS(TBE) vector, that represents not only the most effective, but also the most specific system to downregulate COX-2 in colon cancer cells. Because of the many limits that a retroviral therapy could have in a possible in vivo treatment of CRC, the next goal was to render the enhanced RNAi-mediate COX-2 silencing more suitable for this kind of application. Xiang and et al. (2006) demonstrated that it is possible to induce RNAi in mammalian cells after infection with engineered E. Coli strains expressing Inv and HlyA genes, which encode for two bacterial factors needed for successful transfer of shRNA in mammalian cells. This system, called “trans-kingdom” RNAi (tkRNAi) could represent an optimal approach for the treatment of colorectal cancer, since E. Coli in normally resident in human intestinal flora and could easily vehicled to the tumor tissue. For this reason, I tested the improved COX-2 silencing mediated by pS(COX2) and pS(TBE) vectors by using tkRNAi system. Results obtained in HT-29 and HCA-7 cell lines were in high agreement with data previously collected after the transfection of pS(COX2) and pS(TBE) vectors in the same cell lines. These findings suggest that tkRNAi system for COX-2 silencing, in particular mediated by pS(TBE) vector, could represent a promising tool for the treatment of colorectal cancer. Flanking the studies addressed to the setting-up of a RNAi-mediated therapeutical strategy, I proposed to get ahead with the comprehension of new molecular basis of human colorectal cancer. In particular, it is known that components of the miRNA/RNAi pathway may be altered during the progressive development of colorectal cancer (CRC), and it has been already demonstrated that some miRNAs work as tumor suppressors or oncomiRs in colon cancer. Thus, my hypothesis was that overexpressed COX-2 protein in colon cancer could be the result of decreased levels of one or more tumor suppressor miRNAs. In this thesis, I clearly show an inverse correlation between COX-2 expression and the human miR- 101(1) levels in colon cancer cell lines, tissues and metastases. I also demonstrate that the in vitro modulating of miR-101(1) expression in colon cancer cell lines leads to significant variations in COX-2 expression, and this phenomenon is based on a direct interaction between miR-101(1) and COX-2 mRNA. Moreover, I started to investigate miR-101(1) regulation in the hypoxic environment since adaptation to hypoxia is critical for tumor cell growth and survival and it is known that COX-2 can be induced directly by hypoxia-inducible factor 1 (HIF-1). Surprisingly, I observed that COX-2 overexpression induced by hypoxia is always coupled to a significant decrease of miR-101(1) levels in colon cancer cell lines, suggesting that miR-101(1) regulation could be involved in the adaption of cancer cells to the hypoxic environment that strongly characterize CRC tissues.

Numerical simulations of magma chamber dynamics at Campi Flegrei, and associated seismicity, deformation and gravity changes

Understanding the complex relationships between quantities measured by volcanic monitoring network and shallow magma processes is a crucial headway for the comprehension of volcanic processes and a more realistic evaluation of the associated hazard. This question is very relevant at Campi Flegrei, a volcanic quiescent caldera immediately north-west of Napoli (Italy). The system activity shows a high fumarole release and periodic ground slow movement (bradyseism) with high seismicity. This activity, with the high people density and the presence of military and industrial buildings, makes Campi Flegrei one of the areas with higher volcanic hazard in the world. In such a context my thesis has been focused on magma dynamics due to the refilling of shallow magma chambers, and on the geophysical signals detectable by seismic, deformative and gravimetric monitoring networks that are associated with this phenomenologies. Indeed, the refilling of magma chambers is a process frequently occurring just before a volcanic eruption; therefore, the faculty of identifying this dynamics by means of recorded signal analysis is important to evaluate the short term volcanic hazard. The space-time evolution of dynamics due to injection of new magma in the magma chamber has been studied performing numerical simulations with, and implementing additional features in, the code GALES (Longo et al., 2006), recently developed and still on the upgrade at the Istituto Nazionale di Geofisica e Vulcanologia in Pisa (Italy). GALES is a finite element code based on a physico-mathematical two dimensional, transient model able to treat fluids as multiphase homogeneous mixtures, compressible to incompressible. The fundamental equations of mass, momentum and energy balance are discretised both in time and space using the Galerkin Least-Squares and discontinuity-capturing stabilisation technique. The physical properties of the mixture are computed as a function of local conditions of magma composition, pressure and temperature.The model features enable to study a broad range of phenomenologies characterizing pre and sin-eruptive magma dynamics in a wide domain from the volcanic crater to deep magma feeding zones. The study of displacement field associated with the simulated fluid dynamics has been carried out with a numerical code developed by the Geophysical group at the University College Dublin (O’Brien and Bean, 2004b), with whom we started a very profitable collaboration. In this code, the seismic wave propagation in heterogeneous media with free surface (e.g. the Earth’s surface) is simulated using a discrete elastic lattice where particle interactions are controlled by the Hooke’s law. This method allows to consider medium heterogeneities and complex topography. The initial and boundary conditions for the simulations have been defined within a coordinate project (INGV-DPC 2004-06 V3_2 “Research on active volcanoes, precursors, scenarios, hazard and risk - Campi Flegrei”), to which this thesis contributes, and many researchers experienced on Campi Flegrei in volcanological, seismic, petrological, geochemical fields, etc. collaborate. Numerical simulations of magma and rock dynamis have been coupled as described in the thesis. The first part of the thesis consists of a parametric study aimed at understanding the eect of the presence in magma of carbon dioxide in magma in the convection dynamics. Indeed, the presence of this volatile was relevant in many Campi Flegrei eruptions, including some eruptions commonly considered as reference for a future activity of this volcano. A set of simulations considering an elliptical magma chamber, compositionally uniform, refilled from below by a magma with volatile content equal or dierent from that of the resident magma has been performed. To do this, a multicomponent non-ideal magma saturation model (Papale et al., 2006) that considers the simultaneous presence of CO2 and H2O, has been implemented in GALES. Results show that the presence of CO2 in the incoming magma increases its buoyancy force promoting convection ad mixing. The simulated dynamics produce pressure transients with frequency and amplitude in the sensitivity range of modern geophysical monitoring networks such as the one installed at Campi Flegrei . In the second part, simulations more related with the Campi Flegrei volcanic system have been performed. The simulated system has been defined on the basis of conditions consistent with the bulk of knowledge of Campi Flegrei and in particular of the Agnano-Monte Spina eruption (4100 B.P.), commonly considered as reference for a future high intensity eruption in this area. The magmatic system has been modelled as a long dyke refilling a small shallow magma chamber; magmas with trachytic and phonolitic composition and variable volatile content of H2O and CO2 have been considered. The simulations have been carried out changing the condition of magma injection, the system configuration (magma chamber geometry, dyke size) and the resident and refilling magma composition and volatile content, in order to study the influence of these factors on the simulated dynamics. Simulation results allow to follow each step of the gas-rich magma ascent in the denser magma, highlighting the details of magma convection and mixing. In particular, the presence of more CO2 in the deep magma results in more ecient and faster dynamics. Through this simulations the variation of the gravimetric field has been determined. Afterward, the space-time distribution of stress resulting from numerical simulations have been used as boundary conditions for the simulations of the displacement field imposed by the magmatic dynamics on rocks. The properties of the simulated domain (rock density, P and S wave velocities) have been based on data from literature on active and passive tomographic experiments, obtained through a collaboration with A. Zollo at the Dept. of Physics of the Federici II Univeristy in Napoli. The elasto-dynamics simulations allow to determine the variations of the space-time distribution of deformation and the seismic signal associated with the studied magmatic dynamics. In particular, results show that these dynamics induce deformations similar to those measured at Campi Flegrei and seismic signals with energies concentrated on the typical frequency bands observed in volcanic areas. The present work shows that an approach based on the solution of equations describing the physics of processes within a magmatic fluid and the surrounding rock system is able to recognise and describe the relationships between geophysical signals detectable on the surface and deep magma dynamics. Therefore, the results suggest that the combined study of geophysical data and informations from numerical simulations can allow in a near future a more ecient evaluation of the short term volcanic hazard.

Expression and cellular localization of Copper transporter 2 (Ctr2) in Mus musculus

The Ctr family is an essential part of the copper homeostasis machinery and its members share sequence homology and structural and functional features. Higher eukaryotes express two members of this family Ctr1 and Ctr2. Numerous structural and functional studies are available for Ctr1, the only high affinity Cu(I) transporter thus far identified. Ctr1 holigotrimers mediate cellular copper uptake and this protein was demonstrated to be essential for embryonic development and to play a crucial role in dietary copper acquisition. Instead very little is known about Ctr2, it bears structural homology to the yeast vacuolar copper transporter, which mediates mobilization of vacuolar copper stores. Recent studies using over-expressed epitope-tagged forms of human Ctr2 suggested a function as a low affinity copper transporter that can mediate either copper uptake from the extracellular environment or mobilization of lysosomal copper stores. Using an antibody that recognizes endogenous mouse Ctr2, we studied the expression and localization of endogenous mouse Ctr2 in cell culture and in mouse models to understand its regulation and function in copper homeostasis. By immunoblot we observed a regulation of mCtr2 protein levels in a copper and Ctr1 dependent way. Our observations in cells and transgenic mice suggest that lack of Ctr1 induces a strong downregulation of Ctr2 probably by a post-translational mechanism. By indirect immunofluorescence we observed an exclusive intracellular localization in a perinuclear compartment and no co-localization with lysosomal markers. Immunofluorescence experiments in Ctr1 null cells, supported by sequence analysis, suggest that lysosomes may play a role in mCtr2 biology not as resident compartment, but as a degradation site. In appendix a LC-mass method for analysis of algal biotoxins belonging to the family of PsP (paralytic shellfish poisoning) is described.

Drugstore Bologna. Una lettura del rapporto tra la città e la droga

The main task of this research is to investigate the situation of drugs in the city of Bologna. A first discussion pertains the method to adopt studying an ethical question as drug actually is. In fact it is widely known that drugs problem involves many political and religious considerations which are misleading in a scientific point of view. After a methodological chapter supposed to show the purpose of this research, it is discussed a logical definition of drugs. There it is examined an aristotelian definition of drugs with semantic instruments from philosophy of the language to fulfil meaning of terms. The following chapter discusses personal stories of different people involved in drug in the city, who actually represent the main characters of drug subculture. Afterwards the official statistics concerning drug enforcement is discussed and compared with a specific police action which allows to criticize that data, and to make some hypothesis about drug quantities circulating in town. Next step is investigating drugs addicted in town, with a validation technique of data base queries. The result is a statistics of users in which there is evidence of main presence of foreigners and not resident Italians who use to practice drugs in this city. Demographic analysis of identified people shows that drug addiction is widely diffused among all range of age and mainly pertains males, with an increasing trend. Then is examined the geographic distribution of users residence and use places, showing that drugs abuse is spread among all classes of population, while drugs squares are located in some points of town which realise a kind of drug area with a concentration of dealers not organised together. With some detailed queries in police reports statistics is studied some specific subject on nowadays drug abuse, the phenomenon of multi-use, the relation between drug and crime, the relation between drug and mental disease, recording some evidence in such topics. Finally a survey on city media along last two years shows the interest about this topic and gives an idea of public opinion’s information about drugs. The study refers to the city of Bologna only, and pertains data recorded along last ten years by the local metropolitan police corp.

Vascular wall stem cells. Selection and conditioning of progenitors useful for cell therapy. A pathological case study

The arterial wall contains MSCs with mesengenic and angiogenic abilities. These multipotent precursors have been isolated from variously-sized human adult segments, belying the notion that vessel wall is a relatively quiescent tissue. Recently, our group identified in normal human arteries a vasculogenic niche and subsequently isolated and characterized resident MSCs (VW-MSCs) with angiogenic ability and multilineage potential. To prove that VW-MSCs are involved in normal and pathological vascular remodeling, we used a long-term organ culture system; this method was of critical importance to follow spontaneous 3-D vascular remodeling without any influence of blood cells. Next we tried to identify and localize in situ the VW-MSCs and to understand their role in the vascular remodeling in failed arterial homografts. Subsequently, we isolated this cell population and tested in vitro their multilineage differentiation potential through immunohistochemical, immunofluorescence, RT-PCR and ultrastructural analysis. From 25-30cm2 of each vascular wall homograft sample, we isolated a cell population with MSCs properties; these cells expressed MSC lineage molecules (CD90, CD44, CD105, CD29, CD73), stemness (Notch-1, Oct-4, Sca-1, Stro-1) and pericyte markers (NG2) whilst were negative for hematopoietic and endothelial markers (CD34, CD133, CD45, KDR, CD146, CD31 and vWF). MSCs derived from failed homografts (H-MSCs) exhibited adipogenic, osteogenic and chondrogenic potential but scarce propensity to angiogenic and leiomyogenic differentiation. The present study demonstrates that failed homografts contain MSCs with morphological, phenotypic and functional MSCs properties; H-MSCs are long-lived in culture, highly proliferating and endowed with prompt ability to differentiate into adipocytes, osteocytes and chondrocytes; compared with VW-MSCs from normal arteries, H-MSCs show a failure in angiogenic and leiomyogenic differentiation. A switch in MSCs plasticity could be the basis of pathological remodeling and contribute to aneurysmal failure of arterial homografts. The study of VW-MSCs in a pathological setting indicate that additional mechanisms are involved in vascular diseases; their knowledge will be useful for opening new therapeutic options in cardiovascular diseases.

Valutazione dell'impatto della farmacogenetica in ambito di Sanità Pubblica. Analisi del polimorfismo genetico dell'enzima CYP2D6 metabolizzante xenobiotici.

Pharmacogenetic testing provides an outstanding opportunity to improve prescribing safety and efficacy. In Public health policy pharmacogenetics is relevant for personalized therapy and to maximize therapeutic benefit minimizing adverse events. CYP2D6 is known to be a key enzyme responsible for the biotransformation of about 25-30% of extensively used drugs and genetic variations in genes coding for drug-metabolizing enzymes might lead to adverse drug reactions, toxicity or therapeutic failure of pharmacotherapy. Significant interethnic differences in CYP2D6 allele distribution are well established, but immigration is reshaping the genetic background due to interethnic admixture which introduces variations in individual ancestry resulting in distinct level of population structure. The present thesis deals with the genetic determination of the CYP2D6 alleles actually present in the Emilia-Romagna resident population providing insights into the admixture process. A random sample of 122 natives and 175 immigrants from Africa, Asia and South America where characterized considering the present scenario of migration and back migration events. The results are consistent with the known interethnic genetic variation, but introduction of ethnic specific variants by immigrants predicts a heterogeneous admixed population scenario requiring, for drugs prescription and pharmacogenetics studies, an interdisciplinary approach applied in a properly biogeographical and anthropological frame. To translate pharmacogenetics knowledge into clinical practice requires appropriated public health policies, possibly guiding clinicians to evaluate prospectively which patients have the greatest probability of expressing a variant genotype.

Characterization of vascular wall progenitor cells and their role in therapeutic angiogenesis and Monckeberg's sclerosis

Recently, the existence of a capillary-rich vasculogenic zone has been identified in adult human arteries between the tunica media and adventitia; in this area it has been postulated that Mesenchymal Stem Cells (MSCs) may be present amidst the endothelial progenitors and hematopoietic stem cells. This hypothesis is supported by several studies claiming to have found the in vivo reservoir of MSCs in post-natal vessels and by the presence of ectopic tissues in the pathological artery wall. We demonstrated that the existence of multipotent progenitors is not restricted to microvasculature; vascular wall resident MSCs (VW-MSCs) have been isolated from multidistrict human large and middle size vessels (aortic arch, thoracic aorta and femoral artery) harvested from healthy multiorgan donors. Each VW-MSC population shows characteristics of embryonic-like stem cells and exhibits angiogenic, adipogenic, chondrogenic and leiomyogenic potential but less propensity to osteogenic ifferentiation. Human vascular progenitor cells are also able to engraft, differentiate into mature endothelial cells and support muscle function when injected in a murine model of hind limb ischemia. Conversely, VW-MSCs isolated from calcified femoral arteries display a good response to osteogenic commitment letting us to suppose that VW-MSCs could have an important role in the onset of vascular pathologies such as Mönckeberg sclerosis. Taken together these results show two opposite roles of vascular progenitor cells and underline the importance of establishing their in vivo pathological and regenerative potential to better understand pathological events and promote different therapeutic strategies in cardiovascular research and clinical applications.

Influence of habitat and climate change on European bird communities

Die Fragmentierung von Waldgebieten, der Verlust geeigneter Habitate, die Invasion exotischer Arten und globale Klimaveränderung haben auf Artengemeinschaften erhebliche Auswirkungen. Vögel dienen in vielen Fällen als Indikatorarten für Umweltveränderungen und, besonders, für Veränderungen im Zusammenhang mit globaler Erwärmung. In meiner Arbeit habe ich zuerst einen Literaturüberblick über die Auswirkungen globaler Klimaveränderung auf die Verbreitungsgebiete, den Artenreichtum und die Zusammensetzung von Vogelgemeinschaften dargestellt. Zahlreiche Untersuchungen zeigen, daß die Grenzen der Verbreitungsgebiete der meisten Vogelarten mit klimatischen Faktoren korrelieren. Verschiebungen der Verbreitungsgebiete in nördliche Richtung oder in höhere Regionen im Gebirge konnten bereits für viele temperate Vogelarten beobachtet werden. Weiterhin wurde ein zunehmender Artenreichtum besonders in nördlichen Breiten und in höheren Lagen für viele temperate Vogelgemeinschaften vorhergesagt. In trockenen Gebieten ist dagegen mit einer Abnahme des Artenreichtums zu rechnen. Im zweiten Teil meiner Arbeit habe ich untersucht, ob beobachtete Veränderungen in der Zusammensetzung europäischer Vogelgemeinschaften tatsächlich durch aktuelle Klimaveränderungen beeinflußt werden. Das Zugverhalten der Arten war dabei ein Schwerpunkt der Untersuchung, weil zu erwarten war, daß Vogelarten mit verschiedenem Zugverhalten unterschiedlich auf Klimaveränderungen reagieren. Ich habe ein Regressionsmodell genutzt, welches die räumliche Beziehung zwischen dem Anteil von Langstreckenziehern, Kurzstreckenziehern und Standvögeln in europäischen Vogelgemeinschaften und verschiedenen Klimavariablen beschreibt. Für 21 Gebiete in Europa habe ich Daten über beobachtete Veränderungen in der Struktur der Vogelgemeinschaften und isochrone Klimaveränderungen zusammengetragen. Mit Hilfe dieser Klimaveränderungen und dem räumlichen Regressionsmodell konnte ich berechnen, welche Veränderungen in den Vogelgemeinschaften aufgrund der veränderten Klimabedingungen zu erwarten wären und sie mit beobachteten Veränderungen vergleichen. Beobachtete und berechnete Veränderungen korrelierten signifikant miteinander. Die beobachteten Veränderungen konnten nicht durch räumliche Autokorrelationseffekte oder durch alternative Faktoren, wie z.B. Veränderungen in der Landnutzung, erklärt werden. Im dritten Teil der Arbeit untersuchte ich für eine mitteleuropäische Vogelgemeinschaft welchen Einfluß Habitatveränderungen, die Invasion exotischer Arten und die Klimaveränderung auf Veränderungen der Häufigkeit und Verbreitungsgröße der 159 Vogelarten am Bodensee zwischen 1980-1981 und 2000-2002 hatten. Dabei konnte gezeigt werden, daß Veränderungen in der regionalen Abundanz sowohl durch Habitatveränderungen als auch durch Klimavänderungen hervorgerufen wurden. Exotische Arten schienen in dieser Zeit keinen bedeutenden Einfluß zu haben. Besonders bei Agrarlandarten, Arten mit nördlicheren Verbreitungsgebieten und bei Langstreckenziehern konnten signifikante Abnahmen in der Abundanz beobachtet werden. Vor allem die anhaltenden negativen Bestandsveränderungen bei Langstreckenziehern und die in den letzten zehn Jahren aufgetretenen Abnahmen nördlicher verbreiteter Vogelarten deuten darauf hin, daß die Klimaveränderung aktuell als der größte Einfluß für Vögel in Europa angesehen werden muß. Insgesamt zeigen die Ergebnisse dieser Arbeit, daß sich der anhaltende Druck auf die Umwelt in erster Linie durch Habitat- und Klimaveränderungen manifestiert.

Distacco di retina e lavoro manuale: utilizzo delle schede di dimissione ospedaliera per uno studio di incidenza

Introduzione. La movimentazione manuale di carichi è stata recentemente proposta come un possibile determinante del distacco di retina. Al fine di confortare quest’ipotesi, sono stati analizzati i tassi di incidenza di distacco di retina regmatogeno (DRR) idiopatico, trattato chirurgicamente, tra i residenti in Toscana addetti ad attività lavorative manuali, non manuali e casalinghe. Metodi. Le schede di dimissione ospedaliera (SDO) della Toscana contengono anche informazioni codificate sulla categoria generica di impiego. Sono stati utilizzati i dati di tutti i pazienti residenti in Toscana con una SDO emessa da un qualsiasi ospedale italiano nel periodo 1997-2009, con diagnosi principale di DRR (ICD-9: 361,0-361,07 e 361,9) e con DRG 36 (“interventi sulla retina”). Dopo l’eliminazione dei soggetti che non soddisfacevano i criteri di eligibilità, è stato deciso di restringere la popolazione in studio ai soggetti di età 25-59 anni, successivamente classificati in addetti ad attività lavorative manuali, non manuali o casalinghe. Risultati. Sono stati identificati 1.946 casi. Tra gli uomini, gli addetti ad attività lavorative manuali hanno riportato un tasso di incidenza standardizzato per età 1,8 volte più alto rispetto agli addetti ad attività lavorative non manuali (17,4 [IC95%, 16,1–18,7] vs. 9,8 [IC95%, 8,8–10,8]). Tra le donne, i tassi di incidenza standardizzati per età erano 1,9 volte più alti negli addetti ad attività lavorative manuali (11,1 [IC95%, 9,8–12,3]) e 1,7 volte più alti nelle casalinghe (9,5 [IC95%, 8,3–10,8]) rispetto agli addetti ad attività lavorative non manuali (5,7 [IC95%, 4,8–6,6]). Conclusioni. Lo studio mette in evidenza come gli addetti ad attività lavorative manuali siano maggiormente affetti da DRR idiopatico rispetto agli addetti ad attività lavorative non manuali. Questi risultati supportano l’ipotesi che la movimentazione manuale di carichi, che difficilmente può ritrovarsi come compito di attività lavorative non manuali, possa avere un ruolo causale nella genesi della patologia.

Induktion und Analyse spezifischer Immunantworten nach intranodaler Immunisierung mit RNA im murinen Modell

Die Wirksamkeit einer Vakzine ist von vielen Parametern abhängig. Dazu gehören unter anderen: das ausgewählte Antigen, die Formulation in der das Antigen benutzt wird sowie die Applikationsroute. Antigen-kodierende Ribonukleinsäuren (RNA) gilt heutzutage als eine sichere und effiziente Alternative zu traditionellen Impfstoff-Formulierungen, wie Peptiden, rekombinanten Proteinen, viralen Systemen oder DNA basierten Impfstoffen. Bezüglich des Applikationsortes repräsentiert der Lymphknoten ein optimales Milieu für die Interaktion zwischen antigenpräsentierenden Zellen und T-Zellen. Vor diesem Hintergrund war die Zielsetzung dieser Arbeit, ein auf direktem in vivo Transfer von Antigen-kodierender in vitro transkribierter RNA (IVT-RNA) basierendes Impfverfahren zu entwickeln, zu charakterisieren und auf seine anti-tumorale Wirksamkeit zu testen. In der vorliegenden Arbeit konnte gezeigt werden, dass dendritische Zellen (DCs) in vitro hocheffizient mit IVT-RNA transfiziert werden können und eine hohe stimulatorische Kapazität besitzen. Durch Sequenzmodifikation der IVT-RNA konnten wir die Transkriptstabilität und Translationseffizienz erhöhen was zu einer Steigerung der stimulatorischen Kapazität in vivo führte. Darüber hinaus untersuchten wir die Auswirkung der Insertion eines Signalpeptides 5’ sowie einer C-terminalen transmembran- und zytosolischen-Domäne eines MHC-Klasse-I-Moleküls am 3’ der Antigen-kodierenden Sequenz auf die Effizienz der MHC-Klasse-I und -II Präsentation. Wir konnten in vitro und in vivo nachweisen, dass diese Modifikation zu einer gesteigerten, simultanen Stimulation von antigenspezifischen CD4+ und CD8+ T-Zellen führt. Auf der Basis der optimierten Vektorkassetten etablierten wir die intranodale (i.n.) Transfektion von antigenpräsentierenden Zellen in der Maus. Dazu nutzten wir verschiedene Reportersysteme (eGFP-RNA, fluoreszensmarkierte RNA) und konnten zeigen, dass die intranodale Applikation von IVT-RNA zu selektiven Transfektion und Maturation lymphknotenresidenter DCs führt. Zur Untersuchung der immunologischen Effekte wurden in erster Linie auf Influenza-Hemagglutinin-A und Ovalbumin basierende Modellantigensysteme verwendet. Beide Antigene wurden als Antigen-MHC-Fusionskonstrukte genutzt. Als Responderzellen wurden TCR-transgene Lymphozyten verwendet, die MHC-Klasse-I oder -Klasse-II restringierte Epitope des Influenza-Hemagglutinin-A bzw. des Ovalbumin-Proteins erkennen. Wir konnten in vivo zeigen, dass die intranodale Immunisierung mit IVT-RNA zu einer effizienten Stimulation und Expansion von antigenspezifischen CD4+ und CD8+ T-Zellen in einer dosisabhängigen Weise führt. Funktionell konnte gezeigt werden, dass diese T-Zellen Zytokine sezernieren und zur Zytolyse befähigt sind. Wir waren in der Lage durch repetitive i.n. RNA Immunisierung ein ‚Priming’ CD8+ T-Zellen in naiven Mäusen sowohl gegen virale als auch gegen Tumor assoziierte Antigene zu erreichen. Die geprimten T-Zellen waren befähigt eine zytolytische Aktivität gegen mit spezifischem Peptid beladene Targetzellen zu generieren. Darüber hinaus waren wir in der Lage Gedächtnisszellen expandieren zu können. Abschließend konnten wir in Tumormodellen sowohl in prophylaktischen als auch in therapeutischen Experimenten zeigen dass die i.n. RNA Vakzination die Potenz zur Induktion einer anti-tumoralen Immunität besitzt.