A new PET prototype for proton therapy: comparison of data and Monte Carlo simulations

Ion beam therapy is a valuable method for the treatment of deep-seated and radio-resistant tumors thanks to the favorable depth-dose distribution characterized by the Bragg peak. Hadrontherapy facilities take advantage of the specific ion range, resulting in a highly conformal dose in the target volume, while the dose in critical organs is reduced as compared to photon therapy. The necessity to monitor the delivery precision, i.e. the ion range, is unquestionable, thus different approaches have been investigated, such as the detection of prompt photons or annihilation photons of positron emitter nuclei created during the therapeutic treatment. Based on the measurement of the induced β+ activity, our group has developed various in-beam PET prototypes: the one under test is composed by two planar detector heads, each one consisting of four modules with a total active area of 10 × 10 cm2. A single detector module is made of a LYSO crystal matrix coupled to a position sensitive photomultiplier and is read-out by dedicated frontend electronics. A preliminary data taking was performed at the Italian National Centre for Oncological Hadron Therapy (CNAO, Pavia), using proton beams in the energy range of 93–112 MeV impinging on a plastic phantom. The measured activity profiles are presented and compared with the simulated ones based on the Monte Carlo FLUKA package.

Análisis del comportamiento de uniones metálicas viga-soporte mediante modelo de elementos finitos y comparación de resultados con el modelo de nudos y barras

Para el proyecto y cálculo de estructuras metálicas, fundamentalmente pórticos y celosías de cubierta, la herramienta más comúnmente utilizada son los programas informáticos de nudos y barras. En estos programas se define la geometría y sección de las barras, cuyas características mecánicas son perfectamente conocidas, y sobre las cuales obtenemos unos resultados de cálculo concretos en cuanto a estados tensionales y de deformación. Sin embargo el otro componente del modelo, los nudos, presenta mucha mayor complejidad a la hora de establecer sus propiedades mecánicas, fundamentalmente su rigidez al giro, así como de obtener unos resultados de estados tensionales y de deformación en los mismos. Esta “ignorancia” sobre el comportamiento real de los nudos, se salva generalmente asimilando a los nudos del modelo la condición de rígidos o articulados. Si bien los programas de cálculo ofrecen la posibilidad de introducir nudos con una rigidez intermedia (nudos semirrígidos), la rigidez de cada nudo dependerá de la geometría real de la unión, lo cual, dada la gran variedad de geometrías de uniones que en cualquier proyecto se nos presentan, hace prácticamente inviable introducir los coeficientes correspondientes a cada nudo en los modelos de nudos y barras. Tanto el Eurocódigo como el CTE, establecen que cada unión tendrá asociada una curva momento-rotación característica, que deberá ser determinada por los proyectistas mediante herramientas de cálculo o procedimientos experimentales. No obstante, este es un planteamiento difícil de llevar a cabo para cada proyecto. La consecuencia de esto es, que en la práctica, se realizan extensas comprobaciones y justificaciones de cálculo para las barras de las estructuras, dejando en manos de la práctica común la solución y puesta en obra de las uniones, quedando sin justificar ni comprobar la seguridad y el comportamiento real de estas. Otro aspecto que conlleva la falta de caracterización de las uniones, es que desconocemos como afecta el comportamiento real de éstas en los estados tensionales y de deformación de las barras que acometen a ellas, dudas que con frecuencia nos asaltan, no sólo en la fase de proyecto, sino también a la hora de resolver los problemas de ejecución que inevitablemente se nos presentan en el desarrollo de las obras. El cálculo mediante el método de los elementos finitos, es una herramienta que nos permite introducir la geometría real de perfiles y uniones, y nos permite por tanto abordar el comportamiento real de las uniones, y que está condicionado por su geometría. Por ejemplo, un caso típico es el de la unión de una viga a una placa o a un soporte soldando sólo el alma. Es habitual asimilar esta unión a una articulación. Sin embargo, el modelo por elementos finitos nos ofrece su comportamiento real, que es intermedio entre articulado y empotrado, ya que se transmite un momento y el giro es menor que el del apoyo simple. No obstante, la aplicación del modelo de elementos finitos, introduciendo la geometría de todos los elementos estructurales de un entramado metálico, tampoco resulta en general viable desde un punto de vista práctico, dado que requiere invertir mucho tiempo en comparación con el aumento de precisión que obtenemos respecto a los programas de nudos y barras, mucho más rápidos en la fase de modelización de la estructura. En esta tesis se ha abordado, mediante la modelización por elementos finitos, la resolución de una serie de casos tipo representativos de las uniones más comúnmente ejecutadas en obras de edificación, como son las uniones viga-pilar, estableciendo el comportamiento de estas uniones en función de las variables que comúnmente se presentan, y que son: •Ejecución de uniones viga-pilar soldando solo por el alma (unión por el alma), o bien soldando la viga al pilar por todo su perímetro (unión total). •Disposición o no de rigidizadores en los pilares •Uso de pilares de sección 2UPN en cajón o de tipo HEB, que son los tipos de soporte utilizados en casi el 100% de los casos en edificación. Para establecer la influencia de estas variables en el comportamiento de las uniones, y su repercusión en las vigas, se ha realizado un análisis comparativo entre las variables de resultado de los casos estudiados:•Estados tensionales en vigas y uniones. •Momentos en extremo de vigas •Giros totales y relativos en nudos. •Flechas. Otro de los aspectos que nos permite analizar la casuística planteada, es la valoración, desde un punto de vista de costos de ejecución, de la realización de uniones por todo el perímetro frente a las uniones por el alma, o de la disposición o no de rigidizadores en las uniones por todo el perímetro. Los resultados a este respecto, son estrictamente desde un punto de vista económico, sin perjuicio de que la seguridad o las preferencias de los proyectistas aconsejen una solución determinada. Finalmente, un tercer aspecto que nos ha permitido abordar el estudio planteado, es la comparación de resultados que se obtienen por el método de los elementos finitos, más próximos a la realidad, ya que se tiene en cuenta los giros relativos en las uniones, frente a los resultados obtenidos con programas de nudos y barras. De esta forma, podemos seguir usando el modelo de nudos y barras, más versátil y rápido, pero conociendo cuáles son sus limitaciones, y en qué aspectos y en qué medida, debemos ponderar sus resultados. En el último apartado de la tesis se apuntan una serie de temas sobre los que sería interesante profundizar en posteriores estudios, mediante modelos de elementos finitos, con el objeto de conocer mejor el comportamiento de las uniones estructurales metálicas, en aspectos que no se pueden abordar con los programas de nudos y barras. For the project and calculation of steel structures, mainly building frames and cover lattices, the tool more commonly used are the node and bars model computer programs. In these programs we define the geometry and section of the bars, whose mechanical characteristics are perfectly known, and from which we obtain the all calculation results of stresses and displacements. Nevertheless, the other component of the model, the nodes, are much more difficulty for establishing their mechanical properties, mainly the rotation fixity coefficients, as well as the stresses and displacements. This "ignorance" about the real performance of the nodes, is commonly saved by assimilating to them the condition of fixed or articulated. Though the calculation programs offer the possibility to introducing nodes with an intermediate fixity (half-fixed nodes), the fixity of every node will depend on the real connection’s geometry, which, given the great variety of connections geometries that in a project exist, makes practically unviable to introduce the coefficients corresponding to every node in the models of nodes and bars. Both Eurocode and the CTE, establish that every connection will have a typical moment-rotation associated curve, which will have to be determined for the designers by calculation tools or experimental procedures. Nevertheless, this one is an exposition difficult to carry out for each project. The consequence of this, is that in the practice, in projects are extensive checking and calculation reports about the bars of the structures, trusting in hands of the common practice the solution and execution of the connections, resulting without justification and verification their safety and their real behaviour. Another aspect that carries the lack of the connections characterization, is that we don´t know how affects the connections real behaviour in the stresses and displacements of the bars that attack them, doubts that often assault us, not only in the project phase, but also at the moment of solving the execution problems that inevitably happen in the development of the construction works. The calculation by finite element model is a tool that allows us to introduce the real profiles and connections geometry, and allows us to know about the real behaviour of the connections, which is determined by their geometry. Typical example is a beam-plate or beam-support connection welding only by the web. It is usual to assimilate this connection to an articulation or simple support. Nevertheless, the finite element model determines its real performance, which is between articulated and fixed, since a moment is transmitted and the relative rotation is less than the articulation’s rotation. Nevertheless, the application of the finite element model, introducing the geometry of all the structural elements of a metallic structure, does not also turn out to be viable from a practical point of view, provided that it needs to invest a lot of time in comparison with the precision increase that we obtain opposite the node and bars programs, which are much more faster in the structure modelling phase. In this thesis it has been approached, by finite element modelling, the resolution of a representative type cases of the connections commonly used in works of building, since are the beam-support connections, establishing the performance of these connections depending on the variables that commonly are present, which are: •Execution of beam-support connections welding only the web, or welding the beam to the support for the whole perimeter. •Disposition of stiffeners in the supports •Use 2UPN in box section or HEB section, which are the support types used in almost 100% building cases. To establish the influence of these variables in the connections performance, and the repercussion in the beams, a comparative analyse has been made with the resulting variables of the studied cases: •Stresses states in beams and connections. •Bending moments in beam ends. •Total and relative rotations in nodes. •Deflections in beams. Another aspect that the study allows us to analyze, is the valuation, from a costs point of view, of the execution of connections for the whole perimeter opposite to the web connections, or the execution of stiffeners. The results of this analyse, are strictly from an economic point of view, without prejudice that the safety or the preferences of the designers advise a certain solution. Finally, the third aspect that the study has allowed us to approach, is the comparison of the results that are obtained by the finite element model, nearer to the real behaviour, since the relative rotations in the connections are known, opposite to the results obtained with nodes and bars programs. So that, we can use the nodes and bars models, more versatile and quick, but knowing which are its limitations, and in which aspects and measures, we must weight the results. In the last part of the tesis, are relationated some of the topics on which it would be interesting to approach in later studies, with finite elements models, in order to know better the behaviour of the structural steel connections, in aspects that cannot be approached by the nodes and bars programs.

A genetic selection for Caenorhabditis elegans synaptic transmission mutants.

We have isolated 165 Caenorhabditis elegans mutants, representing 21 genes, that are resistant to inhibitors of cholinesterase (Ric mutants). Since mutations in 20 of the genes appear not to affect acetylcholine reception, we suggest that reduced acetylcholine release contributes to the Ric phenotype of most Ric mutants. Mutations in 15 of the genes lead to defects in a gamma-aminobutyric acid-dependent behavior; these genes are likely to encode proteins with general, rather than cholinergic-specific, roles in synaptic transmission. Ten of the genes have been cloned. Seven encode homologs of proteins that function in the synaptic vesicle cycle: two encode cholinergic-specific proteins, while five encode general presynaptic proteins. Two other Ric genes encode homologs of G-protein signaling molecules. Our assessment of synaptic function in Ric mutants, combined with the homologies of some Ric mutants to presynaptic proteins, suggests that the analysis of Ric genes will continue to yield insights into the regulation and functioning of synapses.

Rapid preparation of giant unilamellar vesicles.

We report here a rapid evaporation method that produces in high yield giant unilamellar vesicles up to 50 microns in diameter. The vesicles are obtained after only 2 min and can be prepared from different phospholipids, including L-alpha-phosphatidylcholine (lecithin), dipalmitoleoyl L-alpha-phosphatidylcholine, and beta-arachidonoyl gamma-palmitoyl L-alpha-phosphatidylcholine. Vesicles can be produced in distilled water and in Hepes, phosphate, and borate buffers in the pH range of 7.0 to 11.5 with ionic strengths up to 50 mM. The short preparation time allows encapsulation of labile molecular targets or enzymes with high catalytic activities. Cell-sized proteoliposomes have been prepared in which gamma-glutamyltransferase (EC 2.3.2.2) was functionally incorporated into the membrane wall.

The structure of bovine F1-ATPase complexed with the peptide antibiotic efrapeptin.

In the previously determined structure of mitochondrial F1-ATPase determined with crystals grown in the presence of adenylyl-imidodiphosphate (AMP-PNP) and ADP, the three catalytic beta-subunits have different conformations and nucleotide occupancies. AMP-PNP and ADP are bound to subunits beta TP and beta DP, respectively, and the third beta-subunit (beta E) has no bound nucleotide. The efrapeptins are a closely related family of modified linear peptides containing 15 amino acids that inhibit both ATP synthesis and hydrolysis by binding to the F1 catalytic domain of F1F0-ATP synthase. In crystals of F1-ATPase grown in the presence of both nucleotides and inhibitor, efrapeptin is bound to a unique site in the central cavity of the enzyme. Its binding is associated with small structural changes in side chains of F1-ATPase around the binding pocket. Efrapeptin makes hydrophobic contacts with the alpha-helical structure in the gamma-subunit, which traverses the cavity, and with subunit beta E and the two adjacent alpha-subunits. Two intermolecular hydrogen bonds could also form. Intramolecular hydrogen bonds probably help to stabilize efrapeptin's two domains (residues 1-6 and 9-15, respectively), which are connected by a flexible region (beta Ala-7 and Gly-8). Efrapeptin appears to inhibit F1-ATPase by blocking the conversion of subunit beta E to a nucleotide binding conformation, as would be required by an enzyme mechanism involving cyclic interconversion of catalytic sites.

Norepinephrine transporters have channel modes of conduction.

Neurotransmitter transporters couple to existing ion gradients to achieve reuptake of transmitter into presynaptic terminals. For coupled cotransport, substrates and ions cross the membrane in fixed stoichiometry. This is in contrast to ion channels, which carry an arbitrary number of ions depending on the channel open time. Members of the gamma-aminobutyric acid transporter gene family presumably function with fixed stoichiometry in which a set number of ions cotransport with one transmitter molecule. Here we report channel-like events from a presumably fixed stoichiometry [norepinephrine (NE)+, Na+, and Cl-], human NE (hNET) in the gamma-aminobutyric acid transporter gene family. These events are stimulated by NE and by guanethidine, an hNET substrate, and they are blocked by cocaine and the antidepressant desipramine. Voltage-clamp data combined with NE uptake data from these same cells indicate that hNETs have two functional modes of conduction: a classical transporter mode (T-mode) and a novel channel mode (C-mode). Both T-mode and C-mode are gated by the same substrates and antagonized by the same blockers. T-mode is putatively electrogenic because the transmitter and cotransported ions sum to one net charge. However, C-mode carries virtually all of the transmitter-induced current, even though it occurs with low probability. This is because each C-mode opening transports hundreds of charges per event. The existence of a channel mode of conduction in a previously established fixed-stoichiometry transporter suggests the appearance of an aqueous pore through the transporter protein during the transport cycle and may have significance for transporter regulation.

Kinetics of cytokine expression during primary human immunodeficiency virus type 1 infection.

In the present study, we have determined the kinetics of constitutive expression of a panel of cytokines [interleukin (IL) 2, IL-4, IL-6, IL-10, interferon gamma (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha)] in sequential peripheral blood mononuclear cell samples from nine individuals with primary human immunodeficiency virus infection. Expression of IL-2 and IL-4 was barely detected in peripheral blood mononuclear cells. However, substantial levels of IL-2 expression were found in mononuclear cells isolated from lymph node. Expression of IL-6 was detected in only three of nine patients, and IL-6 expression was observed when transition from the acute to the chronic phase had already occurred. Expression of IL-10 and TNF-alpha was consistently observed in all patients tested, and levels of both cytokines were either stable or progressively increased over time. Similar to IL-10 and TNF-alpha, IFN-gamma expression was detected in all patients; however, in five of nine patients, IFN-gamma expression peaked very early during primary infection. The early peak in IFN-gamma expression coincided with oligoclonal expansions of CD8+ T cells in five of six patients, and CD8+ T cells mostly accounted for the expression of this cytokine. These results indicate that high levels of expression of proinflammatory cytokines are associated with primary infection and that the cytokine response during this phase of infection is strongly influenced by oligoclonal expansions of CD8+ T cells.

Potent immunogenicity of the B subunits of Escherichia coli heat-labile enterotoxin: receptor binding is essential and induces differential modulation of lymphocyte subsets.

The importance of receptor binding in the potent immunogenicity of Escherichia coli heat-labile enterotoxin B subunit (EtxB) was tested by comparing its immunogical properties with those of a receptor binding mutant, EtxB(G33D). Subcutaneous immunization of EtxB(G33D) resulted in 160-fold reduction in antibody titer compared with wild-type EtxB, whereas its oral delivery failed to provoke any detectable secretory or serum anti-B subunit responses. Moreover, the two proteins induced strikingly different effects on lymphocyte cultures in vitro. EtxB, in comparison with EtxB(G33D), caused an increase in the proportion of B cells, many of which were activated (CD25+); the complete depletion of CD8+ T cells; an increase in the activation of CD4+ T cells; and an increase in interleukin 2 and a decrease in interferon gamma. These data indicate that EtxB exerts profound effects on immune cells, suggesting that its potent immunogenicity is dependent not only on efficient receptor-mediated uptake, but also on direct receptor-mediated immunomodulation of lymphocyte subsets.

Estudo dos espectros vibracionais de poliacetilenos substituídos

Neste trabalho foram sintetizados a poli-2-etinilpiridina (P2EP), poli-4etinilpiridina (P4EP), o iodeto de poli(2-N-t-butilpiridiniumilacetileno) (P2EPtBu) e a poli-β-etinilnaftaleno (Pβ:EN), os quais são poliacetilenos substituídos. Estes polímeros, juntamente com o cloreto de poli(2-piridínio-2-piridilacetileno) (P2EPH), foram caracterizados por espectroscopia vibracional no infravermelho e Raman. Estes polímeros apresentaram variação na posição das bandas Raman com a energia da radiação excitante - chamada dispersão Raman ou fotosseletividade - da ordem de 10 cm-1, bem inferior ao apresentado pelo poliacetileno (cerca de 60 cm-1). Este deslocamento foi interpretado utilizando-se dois dos modelos existentes para descrever este fenômeno: o Modelo de Modo de Amplitude (AMM) e o Modelo de Coordenada de Conjugação Efetiva (ECCM), os quais fornecem informações sobre a estrutura polimérica e sobre seus níveis eletrônicos. Utilizando-se o AMM foi possível obter informações sobre os níveis eletrônicos excitados de mesma simetria que o estado eletrônico fundamental. Por outro lado, o ECCM, com a ajuda de cálculos DFT, mostrou diferenças na extensão da conjugação e no grau de dimerização entre o P2EP na forma cis e trans e indicou que este polímero apresentava, predominantemente, a estrutura cis, fato este confirmado pelos espectros no infravermelho. A dopagem com I2 provocou efeitos diferentes na estrutura dos polímeros. Os espectros no infravermelho dos polímeros dopados indicaram que o P2EP e o P2EPH apresentaram aumento na quantidade de segmentos cis enquanto o P2EPtBu apresentou diminuição na quantidade desses segmentos. Os espectros Raman dos polímeros dopados confirmaram os dados dos espectros no infravermelho. Esta diferença foi interpretada como sendo devida à diferença no volume do substituinte, pois grupos volumosos favorecem o isômero trans-cisóide onde a distância entre os substituintes é maior. A dopagem também levou a um aumento na condutividade dos polímeros, porém os valores de condutividade obtidos foram bem inferiores que os apresentados pelo poliacetileno dopado (10-5 a 10-7 contra 102 S cm-1, tipicamente).

NO adsorption on Pt (111)/Bi surfaces

The adsorption of nitric oxide (NO) on a Pt (111) surface modified with irreversible adsorbed bismuth adatoms is reported. While the voltammetric results reveal a close interaction between the two co-adsorbed compounds. In-situ infrared spectroscopy and scanning tunnelling microscopy indicate the formation of segregated adlayers. Formation of compressed Bi adlayers with modified redox properties is proposed to reconcile both results. This agrees with the observation of Bi islands in the STM images when NO is coadsorbed, not observed in the absence of NO.

Study of dopamine reactivity on platinum single crystal electrode surfaces

Dopamine is the biological molecule responsible, among other functions, of the heart beat and blood pressure regulation. Its loss, in the human body, can result in serious diseases such as Parkinson's, schizophrenia or depression. Structurally, this molecule belongs to the group of catecholamines, together with epinephrine (adrenaline) and norepinephrine (noradrenaline). The hydroquinone moiety of the molecule can be easily oxidized to quinone, rendering the electrochemical methods a convenient approach for the development of dopamine biosensors. The reactivity of similar aromatic molecules, such as catechol and hydroquinone, at well-ordered platinum surfaces, has recently been investigated in our group. In this paper, we extend these studies to the structurally related molecule dopamine. The study has been performed in neutral pH, since this is closer to the natural conditions for these molecules in biological media. Cyclic voltammetry and in situ infra-red spectroscopy have been combined to extract information about the behavior of this molecule on well-defined platinum surfaces. Dopamine appears to be electrochemically active and reveals interesting adsorption phenomena at low potentials (0.15–0.25 V vs RHE), sensitive to the single crystal orientation. The adsorption of dopamine on these surfaces is very strong, taking place at much lower potentials than the electron transfer from solution species. Specifically, the voltammetry of Pt(1 1 1) and Pt(1 0 0) in dopamine solutions shows an oxidation peak at potentials close to the onset of hydrogen evolution, which is related to the desorption of hydrogen and the adsorption of dopamine. On the other hand, adsorption on Pt(1 1 0) is irreversible and the surface appears totally blocked. Spectroscopic results indicate that dopamine is adsorbed flat on the surface. At potentials higher than 0.6 V vs RHE the three basal planes show a common redox process. The initial formation of the quinone moiety is followed by a chemical step resulting in the formation of 5,6-dihydroxyindoline quinone as final product. This oxidation process has also been investigated by vibrational spectroscopy.

Phylogenetic and physiological characterization of a heterotrophic, chemolithoautotrophic Thiothrix strain isolated from activated sludge

The sheathed filamentous bacterium known as strain CT3, isolated by micromanipulation from an activated sludge treatment plant in Italy, is a member of the genus Thiothrix in the gamma-Proteobacteria according to 16S rDNA sequence analysis. The closest phylogenetic neighbours of strain CT3 are strains I and Q(T), which were also isolated from activated sludge and belong to the species Thiothrix fructosivorans. These strains have respectively 99.2 and 99.4 % similarity to CT3 by 16S rDNA sequence comparison. CT3 shows 63-67 % DNA-DNA hybridization with strain I, which is the only currently viable strain of T. fructosivorans. CT3 is the second strain in the genus Thiothrix that has been shown to be capable of growing autotrophically with reduced sulfur compounds as the sole energy source; autotrophy was also confirmed in strain I. The first reported chemolithoautotrophic isolate of this genus was a strain of 'Thiothrix ramosa' that was isolated from a hydrogen sulfide spring and is morphologically distinguishable from all other described strains of Thiothrix, including CT3. CT3 is an aerobic organism that is non-fermentative, not capable of denitrification and able to grow heterotrophically. Autotrophy in the genus Thiothrix should be investigated more fully to better define the taxonomy of this genus.