Porosity and sealing ability of root fillings with gutta-percha and BioRoot RCS or AH Plus sealers. Evaluation by three ex vivo methods

To investigate the ability of BioRoot RCS, a tricalcium silicate-based root canal sealer and AH Plus to effectively fill the root canals of contralateral teeth using three evaluation methods, and to investigate also the correlation between the methods. The prepared root canals of ten pairs of contralateral mandibular premolar teeth were filled with gutta-percha and sealer using lateral compaction. The percentage of voids within the root canal was assessed by micro-computed tomography, whilst sealing ability was investigated by fluid transport and leakage of fluorescent microspheres. The interaction of sealer with dentine, and sealer penetration were assessed by confocal microscopy. The void volume, fluid flow, microsphere leakage and sealer interaction with dentine for both materials were compared. Nonparametric (Mann-Whitney) tests were used to compare the % void and fluid transport of the two sealers. Spearman correlation was used to assess the pairwise relationships between the techniques. The level of significance was set to 0.05. BioRoot RCS exhibited significantly more percentage of voids than AH Plus. There was no difference in fluid flow and microsphere penetration. BioRoot RCS exhibited a different pattern of sealer penetration and interaction with the dentine walls compared to AH Plus. For both materials, the pairwise correlations between the three techniques were close to zero, indicating weak relationships. MicroCT analysis revealed a higher void volume for BioRoot RCS. The other techniques did not show a difference between the sealing ability of the sealers. The correlation between the three ex vivo methods of assessment was weak demonstrating their complementarity rather than their concordance.

Adrenomedullin induces pulmonary vasodilation but does not attenuate pulmonary hypertension in a sheep model of acute pulmonary embolism

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito de diferentes frações inspiradas de oxigênio sobre a ação anti-hipertensiva pulmonar do sildenafil

Pós-graduação em Biotecnologia Animal - FMVZ

Efeitos da adrenomodulina no tratamento da hipertensão pulmonar induzida por microesferas em ovinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Pre- and Posttransplant Monitoring of Alloantibodies by Complement-Dependent Cytotoxicity and Luminex Methodologies in Liver Transplantation

Background. This study evaluated the influence of circulating anti-HLA antibodies on outcomes of 97 liver allografts from deceased donors. Methods. Human leukocyte antigen (HLA) antibody screening was performed by both complement-dependent cytotoxicity (CDC) and multiparameter Luminex microsphere-based assays (Luminex assay). Results. The agreements between T- and B- cell CDC and Luminex assays were 67% and 77% for pre- and posttransplant specimens, respectively. Graft dysfunction was not associated with either positive pretransplant CDC or Luminex panel-reactive antibody (PRA) values. Likewise, positive posttransplant T- or B- cell CDC PRA values were not associated with graft dysfunction. In contrast, posttransplant Luminex PRA values were significantly higher among patients with graft dysfunction compared with subjects with good outcomes (P = .017). Conclusion. Posttransplant monitoring of HLA antibodies with Luminex methodology allowed identification of patients at high-risk for poor graft outcomes.

Anomalous diffusion evidenced by particle tracking at high frame rates.

Diffusion is a common phenomenon in nature and generally is associated with a system trying to reach a local or a global equilibrium state, as a result of highly irregular individual particle motion. Therefore it is of fundamental importance in physics, chemistry and biology. Particle tracking in complex fluids can reveal important characteristics of its properties. In living cells, we coat the microbead with a peptide (RGD) that binds to integrin receptors at the plasma membrane, which connects to the CSK. This procedure is based on the hypothesis that the microsphere can move only if the structure where it is attached move as well. Then, the observed trajectory of microbeads is a probe of the cytoskeleton (CSK), which is governed by several factors, including thermal diffusion, pressure gradients, and molecular motors. The possibility of separating the trajectories into passive and active diffusion may give information about the viscoelasticity of the cell structure and molecular motors activity. And also we could analyze the motion via generalized Stokes-Einstein relation, avoiding the use of any active techniques. Usually a 12 to 16 Frames Per Second (FPS) system is used to track the microbeads in cell for about 5 minutes. Several factors make this FPS limitation: camera computer communication, light, computer speed for online analysis among others. Here we used a high quality camera and our own software, developed in C++ and Linux, to reach high FPS. Measurements were conducted with samples for 10£ and 20£ objectives. We performed sequentially images with different intervals, all with 2 ¹s exposure. The sequences of intervals are in milliseconds: 4 5 ms (maximum speed) 14, 25, 50 and 100 FPS. Our preliminary results highlight the difference between passive and active diffusion, since the passive diffusion is represented by a Gaussian in the distribution of displacements of the center of mass of individual beads between consecutive frames. However, the active process, or anomalous diffusion, shows as long tails in the distribution of displacements.

Multiplex suspension array for human anti-carbohydrate antibody profiling

Glycan-binding antibodies form a significant subpopulation of both natural and acquired antibodies and play an important role in various immune processes. They are for example involved in innate immune responses, cancer, autoimmune diseases, and neurological disorders. In the present study, a microsphere-based flow-cytometric immunoassay (suspension array) was applied for multiplexed detection of glycan-binding antibodies in human serum. Several approaches for immobilization of glycoconjugates onto commercially available fluorescent microspheres were compared, and as the result, the design based on coupling of end-biotinylated glycopolymers has been selected. This method requires only minute amounts of glycans, similar to a printed glycan microarray. The resulting glyco-microspheres were used for detection of IgM and IgG antibodies directed against ABO blood group antigens. The possibility of multiplexing this assay was demonstrated with mixtures of microspheres modified with six different ABO related glycans. Multiplexed detection of anti-glycan IgM and IgG correlated well with singleplex assays (Pearson's correlation coefficient r = 0.95-0.99 for sera of different blood groups). The suspension array in singleplex format for A/B trisaccharide, H(di) and Le(x) microspheres corresponded well to the standard ELISA (r > 0.94). Therefore, the described method is promising for rapid, sensitive, and reproducible detection of anti-glycan antibodies in a multiplexed format.

Dielectrophoresis-based analyte separation and analysis

Dielectrophoresis—the tendency of a material of high dielectric permittivity to migrate in an electrical field gradient to a region of maximum field strength—provides an ideal motive force for manipulating small volumes of biological analytes in microfluidic microsystems. The work described in this thesis was based on the hypothesis that dielectrophoresis could be exploited to provide high-resolution cell separations in microsystems as well as a means for the electrically-controllable manipulation of solid supports for molecular analysis. To this end, a dielectrophoretic/gravitational field-flow-fractionation (DEP/G-FFF) system was developed and the separation performance evaluated using various types and sizes of polystyrene microspheres as model particles. It was shown that separation of the polystyrene beads was based on the differences in their effective dielectrophoretic properties. The ability of an improved DEP/G-FFF system to separate genetically identical, but phenotypically dissimilar cell types was demonstrated using mixtures of 6m2 mutant rat kidney cells grown under transforming and non-transforming culture conditions. Additionally, a panel of engineered dielectric microspheres was designed with specific, predetermined dielectrophoretic properties such that their dielectrophoretic behaviors would be controllable and predictable. The fabrication method involved the use of gold-coated polystyrene microsphere cores coated with a self-assembled monolayer of alkanethiol and, optionally, a self-assembled monolayer of phospholipid to form a thin-insulating-shell-over-conductive-interior structure. The successful development of the DEP/G-FFF separation system and the dielectrically engineered microspheres provides proof-of-principle demonstrations of enabling dielectrophoresis-based microsystem technology that should provide powerful new methods for the manipulation, separation and identification of analytes in many diverse fields. ^

Dissolved lithium and silicon through time in microbial dissolution experiments on ODP Leg 209 peridotite

Using peridotite drilled during Ocean Drilling Program Leg 209, a series of enrichment cultures were initiated on board the ship to stimulate microbially enhanced dissolution of olivine. Dissolution was estimated by measured changes in dissolved Li and Si in the media through time (up to 709 days). The results suggest that there was no significant difference between the amounts of dissolved Li and Si in most of the inoculated microbial cultures compared to the control cultures. Alternative explanations for this are that 1. No microbes are living in the culture tubes that can affect the dissolution rates of olivine, 2. The control cultures have microbes effecting the dissolution of olivine as well as the inoculated cultures, 3. Not enough time has passed to build up a large enough microbial population to effect the dissolution of the olivine in the culture tubes, 4. Microbes act to suppress dissolution of olivine instead of enhancing dissolution, and 5. Abiotic dissolution overshadows microbially enhanced dissolution. Further work is required to test these alternatives.

Distribution of biological, anthropogenic, and volcanic constituents in the San Francisco Bay Coastal System

Although conventional sediment parameters (mean grain size, sorting, and skewness) and provenance have typically been used to infer sediment transport pathways, most freshwater, brackish, and marine environments are also characterized by abundant sediment constituents of biological, and possibly anthropogenic and volcanic, origin that can provide additional insight into local sedimentary processes. The biota will be spatially distributed according to its response to environmental parameters such as water temperature, salinity, dissolved oxygen, organic carbon content, grain size, and intensity of currents and tidal flow, whereas the presence of anthropogenic and volcanic constituents will reflect proximity to source areas and whether they are fluvially- or aerially-transported. Because each of these constituents have a unique environmental signature, they are a more precise proxy for that source area than the conventional sedimentary process indicators. This San Francisco Bay Coastal System study demonstrates that by applying a multi-proxy approach, the primary sites of sediment transport can be identified. Many of these sites are far from where the constituents originated, showing that sediment transport is widespread in the region. Although not often used, identifying and interpreting the distribution of naturally-occurring and allochthonous biologic, anthropogenic, and volcanic sediment constituents is a powerful tool to aid in the investigation of sediment transport pathways in other coastal systems.

Estudio microestructural y de los procesos de hidratación de cementos con adiciones

En los últimos años, las sociedades industrializadas han tomado una mayor conciencia sobre el problema que suponen las emisiones indiscriminadas de gases de efecto invernadero a la atmósfera. El hormigón, cuyo principal componente es el cemento, es probablemente el material más utilizado en construcción. En la actualidad, las emisiones globales de CO2 debidas a la combustión del CaCO3 del cemento Pórtland representan entre el 5% y el 10% respecto del total. Estos valores son de gran interés si se considera que el compromiso aceptado al firmar el Protocolo de Kioto es de una reducción del 5% antes del año 2020, sobre el total de gases producidos. El principal objetivo del presente trabajo es el estudio microestructural y de los procesos de hidratación de los cementos con adiciones. Para ello se propone contribuir a la investigación sobre nuevos productos cementicios basados en micropartículas esféricas vítreas que pueden adicionarse al cemento antes del proceso de amasado. Los resultados obtenidos se han contrastado con las adiciones convencionales de más uso en la actualidad. El nuevo material basa su composición en la química del aluminio y el silicio. Al disminuir la cantidad de CaCO3, se contribuye al desarrollo sostenible y a la reducción de emisiones de CO2. La patente creada por el Grupo Cementos Pórtland Valderrivas (GCPV), describe el proceso de producción de las cemesferas (WO 2009/007470, 2010). Los productos que forman la materia prima para la elaboración de las cemesferas son arcillas, calizas, margas o productos o subproductos industriales, que tras su molienda, son fundidos mediante un fluido gaseoso a elevada temperatura (entre 1250ºC y 1600ºC). Este proceso permite obtener un producto final en forma esférica maciza o microesfera, que tras estabilizarse mediante un enfriamiento rápido, consigue una alta vitrificación idónea para su reactividad química, con una mínima superficie específica en relación a su masa. El producto final obtenido presenta prácticamente la finura requerida y no precisa ser molido, lo que reduce las emisiones de CO2 por el ahorro de combustible durante el proceso de molienda. El proceso descrito permite obtener un amplio abanico de materiales cementantes que, no solo pueden dar respuesta a los problemas generados por las emisiones de CO2, sino también a la disponibilidad de materiales en países donde hasta el momento no se puede fabricar cemento debido a la falta de calizas. Complementariamente se ha optimizado el método de cálculo del grado de hidratación a partir de los resultados del ensayo de ATD-TG en base a los modelos de cálculo de Bhatty y Pane. El método propuesto permite interpretar el comportamiento futuro del material a partir de la interpolación numérica de la cantidad de agua químicamente enlazada. La evolución del grado de hidratación tiene una relación directa con el desarrollo de la resistencia mecánica del material. Con el fin de caracterizar los materiales de base cemento, se ha llevado a cabo una amplia campaña experimental en pasta de cemento, mortero y hormigón. La investigación abarca tres niveles: caracterización microestructural, macroestructural y caracterización del comportamiento a largo plazo, fundamentalmente durabilidad. En total se han evaluado ocho adiciones diferentes: cuatro adiciones convencionales y cuatro tipos de cemesferas con diferente composición química. Los ensayos a escala microscópica comprenden la caracterización química, granulométrica y de la superficie específica BET de los materiales anhidros, análisis térmico diferencial y termogravimétrico en pasta de cemento y mortero, resonancia magnética de silicio en pasta de cemento, difracción de rayos X de los materiales anhidros y de las probetas de pasta, microscopía electrónica de barrido con analizador de energía dispersiva por rayos X en pasta y mortero, y porosimetría por intrusión de mercurio en mortero. La caracterización macroscópica del material comprende ensayos de determinación del agua de consistencia normal y de los tiempos de inicio y fin de fraguado en pasta de cemento, ensayos de resistencia mecánica a flexión y compresión en probetas prismáticas de mortero, y ensayos de resistencia a compresión en probetas de hormigón. Para caracterizar la durabilidad se han desarrollado ensayos de determinación del coeficiente de migración de cloruros y ensayos de resistividad eléctrica en probetas de mortero. Todos los ensayos enumerados permiten clarificar el comportamiento de las cemesferas y compararlo con las distintas adiciones de uso convencional. Los resultados reflejan un buen comportamiento resistente y durable de los materiales con adición de cemesferas. La caracterización microscópica refleja su relación con las propiedades mesoscópicas y permite comprender mejor la evolución en los procesos de hidratación de las cemesferas. In recent years industrialised societies have become increasingly aware of the problem posed by indiscriminate emission of greenhouse gases into the atmosphere. Concrete, with a main component being cement, is arguably the most widely used construction material. At present, global emissions of CO2 due to the combustion of CaCO3 from Portland cement represent between 5% and 10% of the total. If the requirement of the Kyoto Protocol of a reduction of 5% of the total gas produced before 2020 is considered, then such values are of significant interest. The main objective of this work is the assessment of the microstructure and the hydration processes of cements with additions. Such an examination proposes research into new cementitious products based on vitreous spherical microparticles that may be added to the cement before the mixing process. The results are compared with the most commonly used conventional additions. The new material bases its composition on the chemistry of aluminium and silicates. By decreasing the amount of CaCO3, it is possible both to contribute to sustainable development and reduce CO2 emissions. The patent created by Grupo Cementos Portland Valderrivas (GCPV) describes the production process of microspheres (WO 2009/007470, 2010). The products that form the raw material for manufacture are clays, lime-stone, marl and industrial products or by-products that melt after being ground and fed into a gaseous fluid at high temperatures (1250°C and 1600°C). This process allows the obtaining of a product with a solid-spherical or micro-spherical shape and which, after being stabilised in a solid state by rapid cooling, obtains a high vitrification suitable for chemical reactivity, having a minimal surface in relation to its mass. Given that the final product has the fineness required, it prevents grinding that reduces CO2 emissions by saving fuel during this process. The process, which allows a wide range of cementitious materials to be obtained, not only addresses the problems caused by CO2 emissions but also enhances the availability of materials in countries that until the time of writing have not produced cement due to a lack of limestone. In addition, the calculation of the degree of hydration from the test results of DTA-TG is optimised and based on Bhatty and Pane calculation models. The proposed method allows prediction of the performance of the material from numerical interpolation of the amount of chemically bound water. The degree of hydration has a direct relationship with the development of material mechanical strength. In order to characterise the cement-based materials, an extensive experimental campaign in cement paste, concrete and mortar is conducted. The research comprises three levels: micro-structural characterisation, macro-structural and long-term behaviour (mainly durability). In total, eight additions are assessed: four conventional additions and four types of microspheres with different chemical compositions. The micro-scale tests include characterisation of chemical composition, particle size distribution and the BET specific surface area of anhydrous material, differential thermal and thermogravimetric analysis in cement paste and mortar, silicon-29 nuclear magnetic resonance in cement paste, X-ray diffraction of the anhydrous materials and paste specimens, scanning of electron microscopy with energy dispersive X-ray analyser in cement paste and mortar, and mercury intrusion porosimetry in mortar. The macroscopic material characterisation entails determination of water demand for normal consistency, and initial and final setting times of cement paste, flexural and compressive mechanical strength tests in prismatic mortar specimens, and compressive strength tests in concrete specimens. Tests for determining the chloride migration coefficient are performed to characterise durability, together with electrical resistivity tests in mortar specimens. All the tests listed allow clarification of the behaviour of the microspheres and comparison with the various additions of conventional use. The results show good resistance and durable behaviour of materials with a microsphere addition. Microscopic characterisation reflects their relationship with mesoscopic properties and provides insights into the hydration processes of the microspheres.

Portosystemic shunting and persistent fetal vascular structures in aryl hydrocarbon receptor-deficient mice

A physiological examination of mice harboring a null allele at the aryl hydrocarbon (Ah) locus revealed that the encoded aryl hydrocarbon receptor plays a role in the resolution of fetal vascular structures during development. Although the aryl hydrocarbon receptor is more commonly studied for its role in regulating xenobiotic metabolism and dioxin toxicity, a developmental role of this protein is supported by the observation that Ah null mice display smaller livers, reduced fecundity, and decreased body weights. Upon investigating the liver phenotype, we found that the decrease in liver size is directly related to a reduction in hepatocyte size. We also found that smaller hepatocyte size is the result of massive portosystemic shunting in null animals. Colloidal carbon uptake and microsphere perfusion studies indicated that 56% of portal blood flow bypasses the liver sinusoids. Latex corrosion casts and angiography demonstrated that shunting is consistent with the existence of a patent ductus venosus in adult animals. Importantly, fetal vascular structures were also observed at other sites. Intravital microscopy demonstrated an immature sinusoidal architecture in the liver and persistent hyaloid arteries in the eyes of adult Ah null mice, whereas corrosion casting experiments described aberrations in kidney vascular patterns.

Electrochemical performance of carbon gels with variable surface chemistry and physics

This study describes the electrochemical characterization of N-doped carbon xerogels in the form of microspheres and of carbon aerogels with varied porosities and surface oxygen complexes. The interfacial capacitance of N-doped carbon xerogels decreased with increased micropore surface area as determined by N2 adsorption at −196 °C. The interfacial capacitance showed a good correlation with the areal NXPS concentration, and the best correlation with the areal concentration of pyrrolic or pyridonic nitrogen functionalities. The gravimetric capacitance decreased with greater xerogel microsphere diameter. The interfacial capacitance of carbon aerogels increased with higher percentage of porosity as determined from particle and true densities. The interfacial capacitance showed a linear relationship with the areal oxygen concentration and with the areal concentrations of CO- and CO2-evolving groups.

The effect of complexation on characteristics and drug release from PLGA microspheres loaded by cyclosporine-cyclodextrin complex

The purpose of this study was to evaluate the effect of cyclosporine (CyA)-cyclodextrin (CD) complex incorporated within PLGA inicrospheres on microsphere characteristics, with particular emphasis on drug release kinetics. For this purpose, microspheres encapsulated with CyA and those loaded by CyA-CD complex were prepared by solvent evaporation and multiple emulsification solvent evaporation methods, respectively. Morphology, size, encapsulation efficiency and drug release pattern from microspheres were evaluated. Also, physicochemical properties of drug inside microspheres were characterized by differential scanning calorimetry (DSC) and infrared spectroscopy (IR) studies. Scanning electron microscopy (SEM) studies showed that microspheres encapsulated with CyA had islands on the microsphere surface but the islands were not seen on the surface of microspheres loaded by complex. Size range varied from 1 to 25 mu m for CyA encapsulated microspheres and 1 to 50 mu m for complex loaded microspheres. The release of CyA was biphasic with an initial more rapid release phase followed by a slower phase but drug release was twice as fast for complex loaded microspheres. IR studies did not indicate any chemical interaction between the components of microspheres and DSC thermograms revealed that CyA was present either in its amorphous state in microspheres or the presence of CyA as an inclusion complex within microspheres loaded by complex. In conclusion, using CyA as an inclusion complex with CD within microspheres can affect microsphere characteristics and drug release and it is possible to modify microsphere properties like drug release by incorporating CDs as complexing agents.