地球化学新方法在隐状矿床找矿预测中的应用研究

With the progress of prospecting, the need for the discovery of blind ore deposits become more and more urgent. To study and find out the method and technology for the discovery of blind and buried ores is now a priority task. New geochemical methods are key technology to discover blind ores. Information of mobile components related to blind ores were extracted using this new methods. These methods were tested and applied based on element' s mobile components migrating and enriched in geophysical-geochemical process. Several kinds of partial extraction techniques have tested based on element' s occurrence in hypergenic zone. Middle-large scale geochemical methods for exploration in forest and swamp have been tested. A serious of methods were tested and applied effetely about evaluation of regional geochemical anomaly, 1:25000 bedrock or soil geochemical methods sampling based on the net in dendritic water system instead of the normal net. 1. Element related with ores can be mobiled to migrate upwards and be absorpted by surface soil. These abnomal components can be concentrated by natural or artificial methods. These trace metalic ions partially exist in dissovlvable ion forms of active state, and partially have been absorbed by Fe-Mn oxide, soil and organic matter in the soil so that a series of reaction such as complex reaction have take place. Employing various partial extraction techniques, metallic ions related with the phase of the blind ores can be extracted, such as the technique of organic complex extraction, Fe-Mn oxide extraction and the extraction technique of metallic ions of various absorption phases. 2.1:200000 regional geochemical evaluation anomaly methods: Advantageous ore-forming areas were selected firstly. Center, concentration, morphological feature, belt of anomaly were choosed then. Geological and geochemical anomalies were combined. And geological and geochemical background information were restrained. Xilekuduke area in Fuyun sheet , Zhaheba area in Qiakuerte sheet, the west-north part in Ertai sheet and Hongshanzui anomaly in Daqiao sheet were selected as target areas, in Alertai, in the north of Xinjiang. in Xilekuduke area, 1:25000 soil geochemical methods sampling based on the net in dendritic water system was carried out. Cu anomaly and copper mineralization were determined in the center area. Au , Cu anomalies and high polarization anomaly were determined in the south part. Prospecting by primary halo and organic complex extraction were used to prognosis blind ore in widely rang outcrop of bedrock. 1:25000 bedrock or soil geochemical methods sampling based on the net in dendritic water system were used in transported overburden outside of mining area. Shallow seismic method and primary halo found a new blind orebody in mining area. A mineralization site was fou and outside of Puziwan gold mine, in the north of Shanxi province. Developing middle-large scale geochemical exploration method is a key technique based 1:200000 regional geochemical exploration. Some conditions were tested as Sampling density , distribution sites of sample, grain size of sample and occurrence of element for exploration. 1:50000 exploration method was advanced to sample clast sediment supplement clast sediment in valley. 1:25000 bedrock or soil geochemical methods sampling based on the net in dendritic water system was applied to sample residual material in A or C horizon. 1:2000 primary or soil halo methods used to check anomalies and determine mineralization. Daliang gold mineralization in the northern Moerdaoga was found appling these methods. Thermomagnetic method was tested in miniqi copper-polymetallic ore. Process methods such as grain size of sample, heated temperature, magnetic separating technique were tested. A suite of Thermomagnetic geochemical method was formed. This method was applied in Xiangshan Cu~Ni deposit which is cover by clast or Gobi in the eastern Xinjiang. Element's content and contrast of anomaly with Thermomagnetic geochemical method were higher than soil anomaly. Susceptibility after samples were heated could be as a assessment conference for anomaly. In some sectors thermo-magnetic Cu, Ni, Ti anomalious were found outside deposits area. There were strong anomal ies response up ore tested by several kind of partial extraction methods include Thermomagnetic, enzyme leach and other partial extractions in Kalatongke Cu-Ni deposit in hungriness area in the northern of Xinjiang. Element's anomalies of meobile were mainly in Fe-Mn oxide and salt. A Copper mineralization site in Xilekuduke anomaly area had been determined. A blind ore was foung by shallow seismic and geochemical method and a mineralization site was found outside this mining area in Puziwan gold deposit in shanxi province. A Gold mineralization site was found by 1:50000 geochemical exploration in Daliang, Inner Mongolia.

On Directional Selectivity in Vertebrate Retina: An Experimental and Computational Study

This thesis describes an investigation of retinal directional selectivity. We show intracellular (whole-cell patch) recordings in turtle retina which indicate that this computation occurs prior to the ganglion cell, and we describe a pre-ganglionic circuit model to account for this and other findings which places the non-linear spatio-temporal filter at individual, oriented amacrine cell dendrites. The key non-linearity is provided by interactions between excitatory and inhibitory synaptic inputs onto the dendrites, and their distal tips provide directionally selective excitatory outputs onto ganglion cells. Detailed simulations of putative cells support this model, given reasonable parameter constraints. The performance of the model also suggests that this computational substructure may be relevant within the dendritic trees of CNS neurons in general.

Imunoterapia Tumoral com Células Dendríticas

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Development and characterization of novel transgenic techniques for the study of neuronal circuitry

Modern neuroscience relies heavily on sophisticated tools that allow us to visualize and manipulate cells with precise spatial and temporal control. Transgenic mouse models, for example, can be used to manipulate cellular activity in order to draw conclusions about the molecular events responsible for the development, maintenance and refinement of healthy and/or diseased neuronal circuits. Although it is fairly well established that circuits respond to activity-dependent competition between neurons, we have yet to understand either the mechanisms underlying these events or the higher-order plasticity that synchronizes entire circuits. In this thesis we aimed to develop and characterize transgenic mouse models that can be used to directly address these outstanding biological questions in different ways. We present SLICK-H, a Cre-expressing mouse line that can achieve drug-inducible, widespread, neuron-specific manipulations in vivo. This model is a clear improvement over existing models because of its particularly strong, widespread, and even distribution pattern that can be tightly controlled in the absence of drug induction. We also present SLICK-V::Ptox, a mouse line that, through expression of the tetanus toxin light chain, allows long-term inhibition of neurotransmission in a small subset (<1%) of fluorescently labeled pyramidal cells. This model, which can be used to study how a silenced cell performs in a wildtype environment, greatly facilitates the in vivo study of activity-dependent competition in the mammalian brain. As an initial application we used this model to show that tetanus toxin-expressing CA1 neurons experience a 15% - 19% decrease in apical dendritic spine density. Finally, we also describe the attempt to create additional Cre-driven mouse lines that would allow conditional alteration of neuronal activity either by hyperpolarization or inhibition of neurotransmission. Overall, the models characterized in this thesis expand upon the wealth of tools available that aim to dissect neuronal circuitry by genetically manipulating neurons in vivo.

Generation and characterisation of biologically active milk-derived protein and peptide fractions

In recent years, extensive research has been carried out on the health benefits of milk proteins and peptides. Biologically active peptides are defined as specific protein fragments which have a positive impact on the physiological functions of the body; such peptides are produced naturally in vivo, but can also be generated by physical and/or chemical processes, enzymatic hydrolysis and/or microbial fermentation. The aims of this thesis were to investigate not only the traditional methods used for the generation of bioactive peptides, but also novel processes such as heat treatment, and the role of indigenous milk proteases, e.g., in mastitic milk, in the production of such peptides. In addition, colostrum was characterised as a source of bioactive proteins and peptides. Firstly, a comprehensive study was carried out on the composition and physical properties of colostrum throughout the early-lactation period. Marked differences in the physico-chemical properties of colostrum compared with milk were observed. Various fractions of colostrum were also tested for their effect on the secretion of pro- and anti-inflammatory cytokines from a macrophage cell line and bone marrow dendritic cells, as well as insulin secretion from a pancreatic beta cell line. A significant reduction in the secretion of the pro-inflammatory cytokines, TNF-α, IL-6, IL-1β and IL-12, a significant increase in the secretion of the anti-inflammatory cytokine, IL-10, as well as a significant increase in insulin secretion were observed for various colostrum fractions. Another study examined the early proteomic changes in the milk of 8 cows in response to infusion with the endotoxin lipopolysaccharide (LPS) at quarter level in a model mastitic system; marked differences in the protein and peptide profile of milk from LPS challenged cows were observed, and a pH 4.6-soluble fraction of this milk was found to cause a substantial induction in the secretion of IL-10 from a murine macrophage cell line. Heat-induced hydrolysis of sodium caseinate was investigated from the dual viewpoints of protein breakdown and peptide formation, and, a peptide fraction produced in this manner was found to cause a significant increase in the secretion of the anti-inflammatory cytokine, IL-10, from a murine macrophage cell line. The effects of sodium caseinate hydrolysed by chymosin on the gut-derived satiety hormone glucagon-like peptide-1 (GLP-1) were investigated; the resulting casein-derived peptides displayed good in vitro and in vivo secretion of GLP-1. Overall, the studies described in this thesis expand on current knowledge and provide good evidence for the use of novel methods for the isolation, generation and characterisation of bioactive proteins and/or peptides.

Direct TLR2 signaling is critical for NK cell activation and function in response to vaccinia viral infection.

Natural killer (NK) cells play an essential role in innate immune control of poxviral infections in vivo. However, the mechanism(s) underlying NK cell activation and function in response to poxviruses remains poorly understood. In a mouse model of infection with vaccinia virus (VV), the most studied member of the poxvirus family, we identified that the Toll-like receptor (TLR) 2-myeloid differentiating factor 88 (MyD88) pathway was critical for the activation of NK cells and the control of VV infection in vivo. We further showed that TLR2 signaling on NK cells, but not on accessory cells such as dendritic cells (DCs), was necessary for NK cell activation and that this intrinsic TLR2-MyD88 signaling pathway was required for NK cell activation and played a critical role in the control of VV infection in vivo. In addition, we showed that the activating receptor NKG2D was also important for efficient NK activation and function, as well as recognition of VV-infected targets. We further demonstrated that VV could directly activate NK cells via TLR2 in the presence of cytokines in vitro and TLR2-MyD88-dependent activation of NK cells by VV was mediated through the phosphatidylinositol 3-kinase (PI3K)-extracellular signal-regulated kinase (ERK) pathway. Taken together, these results represent the first evidence that intrinsic TLR signaling is critical for NK cell activation and function in the control of a viral infection in vivo, indicate that multiple pathways are required for efficient NK cell activation and function in response to VV infection, and may provide important insights into the design of effective strategies to combat poxviral infections.

Primary vascularization of the graft determines the immunodominance of murine minor H antigens during organ transplantation.

Grafts can be rejected even when matched for MHC because of differences in the minor histocompatibility Ags (mH-Ags). H4- and H60-derived epitopes are known as immunodominant mH-Ags in H2(b)-compatible BALB.B to C57BL/6 transplantation settings. Although multiple explanations have been provided to explain immunodominance of Ags, the role of vascularization of the graft is yet to be determined. In this study, we used heart (vascularized) and skin (nonvascularized) transplantations to determine the role of primary vascularization of the graft. A higher IFN-γ response toward H60 peptide occurs in heart recipients. In contrast, a higher IFN-γ response was generated against H4 peptide in skin transplant recipients. Peptide-loaded tetramer staining revealed a distinct antigenic hierarchy between heart and skin transplantation: H60-specific CD8(+) T cells were the most abundant after heart transplantation, whereas H4-specific CD8(+) T cells were more abundant after skin graft. Neither the tissue-specific distribution of mH-Ags nor the draining lymph node-derived dendritic cells correlated with the observed immunodominance. Interestingly, non-primarily vascularized cardiac allografts mimicked skin grafts in the observed immunodominance, and H60 immunodominance was observed in primarily vascularized skin grafts. However, T cell depletion from the BALB.B donor prior to cardiac allograft induces H4 immunodominance in vascularized cardiac allograft. Collectively, our data suggest that immediate transmigration of donor T cells via primary vascularization is responsible for the immunodominance of H60 mH-Ag in organ and tissue transplantation.

B cells in rheumatoid synovitis.

In rheumatoid arthritis, T cells, B cells, macrophages, and dendritic cells invade the synovial membranes, establishing complex microstructures that promote inflammatory/tissue destructive lesions. B cell involvement has been considered to be limited to autoantibody production. However, recent studies suggest that B cells support rheumatoid disease through other mechanisms. A critical element of rheumatoid synovitis is the process of ectopic lymphoid neogenesis, with highly efficient lymphoid architectures established in a nonlymphoid tissue site. Rheumatoid synovitis recapitulates the pathways of lymph node formation, and B cells play a key role in this process. Furthermore, studies of rheumatoid lesions implanted in immunodeficient mice suggest that T cell activation in synovitis is B cell dependent, indicating the role played by B cells in presenting antigens and providing survival signals.

The effects of thermo-electrically induced convection in alloy solidification

The effects of a constant uniform magnetic field on thermoelectric currents during dendritic solidification were investigated using a 2-dimensional enthalpy based numerical model. Using an approximation of the dendrite growing in free space it was found that the resulting Lorentz force generates a circulating flow influencing the solidification pattern. As the magnetic field strength increases it was found that secondary growth on the clockwise side of the primary arm of the dendrite was encouraged, while the anticlockwise side is suppressed due to a reduction in local free energy. The preferred direction of growth rotated in the clockwise sense under an anti-clockwise flow for both the binary alloy and pure material. The tip velociy is significantly increased compared to growth in stagnant flow. This is due to a small recirculation that follows the tip of the dendrite; bringing in colder liquid and lower concentrations of solute. The recirculation being not normally incident on the tip is most likely the cause for the rotation. Grain growth consisting of multiple seeds with the same anisotropy growing in the same plane, gives a competition to release latent heat resulting in stunted growth. The initial growth for each dendrite is very similar to the single seed cases indicating that dendrites must become before the thermoelectric interactions are significant.

Effects of magnetic fields on crystal growth

The effects of a constant uniform magnetic field on dendritic solidification were investigated using a 2-dimensional enthalpy based numerical model. The interaction between thermoelectic currents and the magnetic field generates a Lorentz force that creates a flow. This flow causes a change in the morphology of the dendrite; secondary growth is promoted on one side of the dendrite arm and the tip velocity of the primary arm is increased.

Thermoelectric effects on alloy solidification microstructure

Thermoelectric currents in the presence of a magnetic field generate Lorentz forces which can drive fluid flow. In the case of dendritic growth a naturally occurring thermoelectric current exists and in the presence of a high magnetic field micro convections are generated. Experimental evidence has attributed changes in microstructure to this effect. A numerical model has been developed to study the flow field around an unconstricted equiaxed dendrite growing under these conditions. The growth is modeled in 2D and 3D by an enthalpy based method and a complex flow structure has been predicted. Using a pseudo-3D approximation for economy, realistic 2D simulations are obtained where a fully coupled transient scheme reveals significant changes to the dendrite morphology reflecting experimental evidence. There is a rotation of the preferred direction of growth and increased secondary branching.

Multiscale modeling of the onset of freckle formation during vacuum arc remelting

A multiscale model for the Vacuum Arc Remelting process (VAR) was developed to simulate dendritic microstructures during solidification and investigate the onset of freckle formation. On the macroscale, a 3D multi-physics model of VAR was used to study complex physical phenomena, including liquid metal flow with turbulence, heat transfer, and magnetohydrodynamics. The results showed that unsteady fluid flow in the liquid pool caused significant thermal perturbation at the solidification front. These results were coupled into a micromodel to simulate dendritic growth controlled by solute diffusion, including local remelting. The changes in Rayleigh number as the microstructure remelts was quantified to provide an indicator of when fluid flow channels (i.e. freckles) will initiate in the mushy zone. By examining the simulated microstructures, it was found that the Rayleigh number increased more than 300 times during remelting, which suggests that thermal perturbation could be responsible for the onset of freckle formation.

Effects of magnetic fields on crystal growth

The effects of a constant uniform magnetic field on thermoelectric currents during dendritic solidification were investigated using an enthalpy based numerical model. It was found that the resulting Lorentz force generates a complex flow influencing the solidification pattern. Experimental work of material processing under high magnetic field conditions has shown that the microstructure can be significantly altered. There is evidence that these effects can be atrtributed to the Lorentz force created through the thermoelectric magentohydrodynamic interactions.[1,2] However the mechanism of how this occurs is not very well understood. In this paper, our aim is to investigate the flow field created from the Lorentz force and how this influences the morphology of dendritic growth for both pure materials and binary alloys.

Effects of magnetic fields on crystal growth

The effects of a constant uniform magnetic field on thermoelectric currents during dendritic solidification were investigated using a two-dimensional enthalpy based numerical model. Using an approximation for three-dimensional unconstricted growth, the resulting Lorentz forces generate a circulating flow influencing the solidification pattern. Under the presence of a strong magnetic field secondary growth on the clockwise side of the primary arm of the dendrite was encouraged, whereas the anticlockwise side is suppressed due to a reduction in local free energy. The preferred direction of growth rotated in the clockwise sense under an anticlockwise flow. The tip velocity is significantly increased compared with growth in stagnant flow. This is due to a small recirculation at the tip of the dendrite; bringing in colder liquid and lowering the concentration of solute.

IMPACT OF RNA VIRUSES ON THE REGULATION OF IL-23 IN MOUSE AND HUMAN MODELS OF INFECTION.

Interluekin-23 (IL-23) is a pro-inflammatory cytokine critical to the regulation of innate and adaptive immune responses. The main role for this cytokine is in the proliferation and differentiation of the IL-17 producing CD4 T helper cell, Th17. Virus infection deregulates IL-23 expression and function, but little is known about the mechanism behind this phenomena. Here, I demonstrate a reduction of Toll like receptor (TLR) ligand-induced IL-23 expression in lymphocytic choriomeningitis virus (LCMV)-infected bone marrow-derived dendritic cells (BMDCs), indicating that a function of these cells is disrupted during virus infection. I propose a mechanism of TLR ligand-induced IL-23 expression inhibition upon LCMV infection via the deactivation of p38, AP-1, and NF-κB. Further analysis revealed a direct relationship between LCMV infection with the IL-10 and SOCS3 expression. To understand IL-23 function, I characterized IL-23-induced JAK/STAT signalling pathway and IL-23 receptor expression on human CD4 T cells. My results demonstrate that IL-23 induces activation of p-JAK2, p-Tyk2, p-STAT1, p-STAT3, and p-STAT4 in CD4 T cells. For the first time I show that IL-23 alone induces the expression of its own receptor components, IL-12Rβ1 and IL-23Rα, in CD4 T cells. Blocking JAK2, STAT1, and STAT3 activation with specific inhibitors detrimentally effected expression of IL-23 receptor demonstrating that activation of JAK/STAT signalling is important for IL-23 receptor expression. I also addressed the effect of viral infection on IL-23 function and receptor expression in CD4 T cells using cells isolated from HIV positive individuals. These studies were based on earlier reports that the expression of IL-23 and the IL-23 receptor are impaired during HIV infection. I demonstrate that the phosphorylation of JAK2, STAT1, and STAT3 induced by IL-23, as well as IL-23 receptor expression are deregulated in CD4 T cells isolated from HIV positive individuals. This study has furthered the understanding of how the expression and function of IL-23 is regulated during viral infections.