Pro-matrix metalloproteinase-2 transfection increases orthotopic primary growth and experimental metastasis of MDA-MB-231 human breast cancer cells in nude mice

The ability to activate pro-matrix metalloproteinase (pro-MMP)-2 via membrane type-MMP is a hallmark of human breast cancer cell lines that show increased invasiveness, suggesting that MMP-2 contributes to human breast cancer progression. To investigate this, we have stably transfected pro-MMP-2 into the human breast cancer cell line MDA-MB-231, which lacks MMP-2 expression but does express its cell surface activator, membrane type 1-MMP. Multiple clones were derived and shown to produce pro-MMP-2 and to activate it in response to concanavalin A. In vitro analysis showed that the pro-MMP-2-transfected clones exhibited an increased invasive potential in Boyden chamber and Matrigel outgrowth assays, compared with the parental cells or those transfected with vector only. When inoculated into the mammary fat pad of nude mice, each of the MMP-2-tranfected clones grew faster than each of the vector controls tested. After intracardiac inoculation into nude mice, pro-MMP-2-transfected clones showed a significant increase in the incidence of metastasis to brain, liver, bone, and kidney compared with the vector control clones but not lung. Increased tumor burden was seen in the primary site and in lung metastases, and a trend toward increased burden was seen in bone, however, no change was seen in brain, liver, or kidney. This data supports a role for MMP-2 in breast cancer progression, both in the growth of primary tumors and in their spread to distant organs. MMP-2 may be a useful target for breast cancer therapy when refinement of MMP inhibitors provides for MMP-specific agents.

Transfection of MDA-MB-231 human breast carcinoma cells with bone sialoprotein (BSP) stimulates migration and invasion in vitro and growth of primary and secondary tumors in nude mice

We have investigated the role of bone sialoprotein (BSP), a secreted glycoprotein normally found in bone, in breast cancer progression. To explore functions for BSP in human breast cancer invasion and metastasis, the full-length BSP cDNA was transfected into the MDA-MB-231-BAG human breast cancer cell line under the control of the CMV promoter. Clones expressing BSP and vector control clones were isolated. BSP producing clones showed increased monolayer wound healing, a faster rate of stellate outgrowth in Matrigel and increased rate of invasion into a collagen matrix when compared to control clones. Clones were also examined in models of breast cancer growth and metastasis in vivo. BSP transfected clones showed an increased rate of primary tumor growth following mammary fat pad injection of nude mice. BSP transfected clones and vector control clones metastasized to soft organs and bone at a similar rate after intra-cardiac injection as determined by real-time PCR and X-ray analysis. Although these organs were targets for both BSP transfected and non-transfected cells, the size of the metastatic lesion was shown to be significantly larger for BSP expressing clones. This was determined by real-time PCR analysis for soft organs and by X-ray analysis of bone lesions. For bone this was confirmed by intra-tibial injections of cells in nude mice. We conclude that BSP acts to drive primary and secondary tumor growth of breast cancers in vivo.

The decline of northern malaria and population dynamics of Plamodium vivax

Europe was declared malaria free in 1975. The disappearance of malaria has traditionally been attributed to numerous deliberate actions like vector control, the screening of houses, more efficient medication etc. Malaria, however, disappeared from many countries like Finland before any counter measures had even started. The aim of this thesis is to study the population ecology of P. vivax and its interaction with the human host and the vector. By finding the factors that attributed to the extinction of vivax malaria it might be possible to improve the modern strategy against P. vivax. The parasite was studied with data from Finland, which provides the longest time series (1749-2008) of malaria statistics in the world. The malaria vectors, Anopheles messeae and A. beklemishevi are still common species in the country. The eradication of vivax malaria is difficult because the parasite has a dormant stage that can cause a relapse long after a primary infection. It was now shown that P. vivax is able to detect the presence of a potential vector. A dormant stage is triggered even from a bite of an uninfected Anopheles mosquito. This optimizes the chances for the Plasmodium to reach a mosquito vector for sexual reproduction. The longevity of the dormant stage could be shown to be at least nine years. The parasite spends several years in its human host and the behaviour of the human carrier had a profound impact on the decline of the disease in Finland. Malaria spring epidemics could be explained by a previous warm summer. Neither annual nor summer mean temperature had any impact on the long term malaria trend. Malaria disappeared slowly from Finland without mosquito control. The sociological change from extended families to nuclear families led to decreased household size. The decreased household size correlated strongly with the decline of malaria. That led to an increased isolation of the subpopulations of P. vivax. Their habitat consisted of the bedrooms in which human carriers slept together with the overwintering vectors. The isolation of the parasite ultimately led to the extinction of vivax malaria. Metapopulation models adapted to local conditions should therefore be implemented as a tool for settlement planning and socio-economic development and become an integrated part of the fight against malaria.

Development of salinity tolerance in rice by constitutive-overexpression of genes involved in the regulation of programmed cell death

Environmental factors contribute to over 70% of crop yield losses worldwide. Of these drought and salinity are the most significant causes of crop yield reduction. Rice is an important staple crop that feeds more than half of the world’s population. However among the agronomically important cereals rice is the most sensitive to salinity. In the present study we show that exogenous expression of anti-apoptotic genes from diverse origins, AtBAG4 (Arabidopsis), Hsp70 (Citrus tristeza virus) and p35 (Baculovirus), significantly improves salinity tolerance in rice at the whole plant level. Physiological, biochemical and agronomical analyses of transgenic rice expressing each of the anti-apoptotic genes subjected to salinity treatment demonstrated traits associated with tolerant varieties including, improved photosynthesis, membrane integrity, ion and ROS maintenance systems, growth rate, and yield components. Moreover, FTIR analysis showed that the chemical composition of salinity-treated transgenic plants is reminiscent of non-treated, unstressed controls. In contrast, wild type and vector control plants displayed hallmark features of stress, including pectin degradation upon subjection to salinity treatment. Interestingly, despite their diverse origins, transgenic plants expressing the anti-apoptotic genes assessed in this study displayed similar physiological and biochemical characteristics during salinity treatment thus providing further evidence that cell death pathways are conserved across broad evolutionary kingdoms. Our results reveal that anti-apoptotic genes facilitate maintenance of metabolic activity at the whole plant level to create favorable conditions for cellular survival. It is these conditions that are crucial and conducive to the plants ability to tolerate/adapt to extreme environments.

Improved prediction of plane transverse jets in supersonic crossflows

THE flowfield due to transverse injection of a round sonic jet into a supersonic flowis a configuration of interest in the design of supersonic combustors or thrust vector control of supersonic jets. The flow is also of fundamental interest because it presents separation from a smooth surface, embedded subsonic regions, curved shear layers, strong shocks, an unusual development of the injected jet into a kidney-shaped streamwise vortex pair, and a wake behind the jet. Although the geometry is simple, the flow is complex and is a good candidate for assessing the behavior of turbulence models for high-speed flow, beginning with the corresponding two-dimensional flow shown in Fig. 1. At the slot, an underexpanded sonic jet expands rapidly into the supersonic crossflow. Expansion waves reflect at the jet boundary, coalesce, and give rise to a Mach surface (Mach disk for round jets).

Defective in Cuticular Ridges (DCR) of Arabidopsis thaliana, a Gene Associated with Surface Cutin Formation, Encodes a Soluble Diacylglycerol Acyltransferase

A key step in the triacylglycerol (TAG) biosynthetic pathway is the final acylation of diacylglycerol (DAG) by DAG acyltransferase. In silico analysis has revealed that the DCR (defective in cuticular ridges) (At5g23940) gene has a typical HX4D acyltransferase motif at the N-terminal end and a lipid binding motif VX(2)GF at the middle of the sequence. To understand the biochemical function, the gene was overexpressed in Escherichia coli, and the purified recombinant protein was found to acylate DAG specifically in an acyl-CoA-dependent manner. Overexpression of At5g23940 in a Saccharomyces cerevisiae quadruple mutant deficient in DAG acyltransferases resulted in TAG accumulation. At5g23940 rescued the growth of this quadruple mutant in the oleate-containing medium, whereas empty vector control did not. Lipid particles were localized in the cytosol of At5g23940-transformed quadruple mutant cells, as observed by oil red O staining. There was an incorporation of 16-hydroxyhexadecanoic acid into TAG in At5g23940-transformed cells of quadruple mutant. Here we report a soluble acyl-CoA-dependent DAG acyltransferase from Arabidopsis thaliana. Taken together, these data suggest that a broad specific DAG acyltransferase may be involved in the cutin as well as in the TAG biosynthesis by supplying hydroxy fatty acid.

A method of tracking the peak power points for a variable speed wind energy conversion system

In this paper, a method of tracking the peak power in a wind energy conversion system (WECS) is proposed, which is independent of the turbine parameters and air density. The algorithm searches for the peak power by varying the speed in the desired direction. The generator is operated in the speed control mode with the speed reference being dynamically modified in accordance with the magnitude and direction of change of active power. The peak power points in the P-omega curve correspond to dP/domega = 0. This fact is made use of in the optimum point search algorithm. The generator considered is a wound rotor induction machine whose stator is connected directly to the grid and the rotor is fed through back-to-back pulse-width-modulation (PWM) converters. Stator flux-oriented vector control is applied to control the active and reactive current loops independently. The turbine characteristics are generated by a dc motor fed from a commercial dc drive. All of the control loops are executed by a single-chip digital signal processor (DSP) controller TMS320F240. Experimental results show that the performance of the control algorithm compares well with the conventional torque control method.

Apparatus and method for heat-run test on high-power PWM converters with low energy expenditure

Before installation, a voltage source converter is usually subjected to heat-run test to verify its thermal design and performance under load. For heat-run test, the converter needs to be operated at rated voltage and rated current for a substantial length of time. Hence, such tests consume huge amount of energy in case of high-power converters. Also, the capacities of the source and loads available in the research and development (R&D) centre or the production facility could be inadequate to conduct such tests. This paper proposes a method to conduct heat-run tests on high-power, pulse width modulated (PWM) converters with low energy consumption. The experimental set-up consists of the converter under test and another converter (of similar or higher rating), both connected in parallel on the ac side and open on the dc side. Vector-control or synchronous reference frame control is employed to control the converters such that one draws certain amount of reactive power and the other supplies the same; only the system losses are drawn from the mains. The performance of the controller is validated through simulation and experiments. Experimental results, pertaining to heat-run tests on a high-power PWM converter, are presented at power levels of 25 kVA to 150 kVA.

Harmonic Current Injection based Powerline Communication Method for Grid Connected Single Phase Inverters in Domestic Microgrids

The following paper presents a Powerline Communication (PLC) Method for Single Phase interfaced inverters in domestic microgrids. The PLC method is based on the injection of a repeating sequence of a specific harmonic, which is then modulated on the fundamental component of the grid current supplied by the inverters to the microgrid. The power flow and information exchange are simultaneously accomplished by the grid interacting inverters based on current programmed vector control, hence there is no need for dedicated hardware. Simulation results have been shown for inter-inverter communication under different operating conditions to propose the viability. These simulations have been experimentally validated and the corresponding results have also been presented in the paper.

A Mycobacterial Phosphoribosyltransferase Promotes Bacillary Survival by Inhibiting Oxidative Stress and Autophagy Pathways in Macrophages and Zebrafish

Mycobacterium tuberculosis employs various strategies to modulate host immune responses to facilitate its persistence in macrophages. The M. tuberculosis cell wall contains numerous glycoproteins with unknown roles in pathogenesis. Here, by using Concanavalin A and LC-MS analysis, we identified a novel mannosylated glycoprotein phosphoribosyltransferase, encoded by Rv3242c from M. tuberculosis cell walls. Homology modeling, bioinformatic analyses, and an assay of phosphoribosyltransferase activity in Mycobacterium smegmatis expressing recombinant Rv3242c (MsmRv3242c) confirmed the mass spectrometry data. Using Mycobacterium marinum-zebrafish and the surrogate MsmRv3242c infection models, we proved that phosphoribosyltransferase is involved in mycobacterial virulence. Histological and infection assays showed that the M. marinum mimG mutant, an Rv3242c orthologue in a pathogenic M. marinum strain, was strongly attenuated in adult zebrafish and also survived less in macrophages. In contrast, infection with wild type and the complemented Delta mimG: Rv3242c M. marinum strains showed prominent pathological features, such as severe emaciation, skin lesions, hemorrhaging, and more zebrafish death. Similarly, recombinant Msm Rv3242c bacteria showed increased invasion in non-phagocytic epithelial cells and longer intracellular survival in macrophages as compared with wild type and vector control M. smegmatis strains. Further mechanistic studies revealed that the Rv3242c- and mimG-mediated enhancement of intramacrophagic survival was due to inhibition of autophagy, reactive oxygen species, and reduced activities of superoxide dismutase and catalase enzymes. Infection with MsmRv3242c also activated the MAPK pathway, NF-kappa B, and inflammatory cytokines. In summary, we show that a novel mycobacterial mannosylated phosphoribosyltransferase acts as a virulence and immunomodulatory factor, suggesting that it may constitute a novel target for antimycobacterial drugs.

A construção da integralidade nas práticas em saúde: o processo de implantação das ações de vigilância da dengue na estratégia de Saúde da Família do município de Piraí RJ

As atividades de controle da dengue no município de Piraí RJ, incorporadas às atividades da Estratégia de Saúde da Família (ESF) desde 2005, experimentaram avanços relacionados às ações de Vigilância Ambiental em Saúde, envolvidos na vigilância da dengue. O novo modelo promoveu a integração das ações, a vinculação e responsabilidade do setor saúde com a população adscrita, fortalecendo o cuidado com o território na prevenção de doenças e agravos transmitidos por vetores. Esta pesquisa teve como objetivo a análise do processo de implantação da integração das ações de vigilância da dengue nas unidades da Estratégia de Saúde da Família de Piraí tendo como eixo norteador a construção da integralidade nas práticas em saúde. Realizou a contextualização do processo político gerencial que motivou e garantiu a integração das ações e buscou identificar como são as percepções, significados e os desafios pelos Agentes Comunitários de Saúde (ACS) no dia a dia das ações de vigilância e da integração delas com suas práticas cotidianas. Os resultados dos indicadores de controle do vetor, obtidos após a integração das ações no município de Piraí, mostram que ela tem sido efetiva. No grupo focal os agentes de saúde expressaram a dificuldade da população em perceber e entender esse novo papel desempenhado pelos ACS. Por outro lado, eles têm grande preocupação com a possibilidade desta nova atividade vir a interferir no vínculo deles com a comunidade. Também foi identificado que os agentes ainda distanciam a ação de educação em saúde da vigilância ambiental. Esta dificuldade parece estar relacionada a própria concepção, ainda medicalizada e individualista, do papel da educação e promoção em saúde a serem desenvolvidas no território. Apesar do grande sucesso alcançado na integração das ações, é preciso entender que este é um processo dinâmico e em constante transformação. Ele implica em um exercício constante de avaliação e diálogo sobre as questões envolvidas no cotidiano desta prática.

Controle por modo deslizante para sistemas não-lineares com atraso.

Nesta Dissertação são propostos dois esquemas de controle para sistemas não-lineares com atraso. No primeiro, o objetivo é controlar uma classe de sistemas incertos multivariáveis, de grau relativo unitário, com perturbações não-lineares descasadas dependentes do estado, e com atraso incerto e variante no tempo em relação ao estado. No segundo, deseja-se controlar uma classe de sistemas monovariáveis, com parâmetros conhecidos, grau relativo arbitrário, atraso arbitrário conhecido e constante na saída. Admitindo-se que o atraso na entrada pode ser deslocado para a saída, então, o segundo esquema de controle pode ser aplicado a sistemas com atraso na entrada. Os controladores desenvolvidos são baseados no controle por modo deslizante e realimentação de saída, com função de modulação para a amplitude do sinal de controle. Além disso, observadores estimam as variáveis de estado não-medidas. Em ambos os esquemas de controle propostos, garante-se propriedades de estabilidade globais do sistema em malha fechada. Simulações ilustram a eficácia dos controladores desenvolvidos.

永磁伺服电动机的矢量调速控制系统研究

永磁同步伺服电动机（PMSM）以其优越的性能广泛应用于各个机械传动领域。对PMSM的研究具有非常大的实际意义和价值，尤其对于我们单位目前在建大科学工程CSR。本文采用美国TI公司专用于电动机控制的数字信号处理器（DSP）芯片TMs320LF2406A作为核心，设计和开发全数字化的PMSM矢量调速控制系统。深入研究永磁同步电动机的矢量控制理论，建立起相应数学模型，然后提出矢量控制调速方案，并通过Mat lab仿真论证其可行性。介绍了硬件，软件结构及其实现。硬件方面主要介绍了控制电路各部分的设计和调试。在硬件基础上，用T工公司DSP汇编语言编程，实现电流环和速度环的双环控制，给出了系统程序和PWM信号产生的思路，并给出了主要模块的源程序。为了提高程序的运行效率，节省存储空间和提高系统的可靠性，程序中尽可能多的采用了数据表结构。

基于神经网络的永磁同步电机矢量控制

采用单神经元自适应PID控制器对永磁同步电机进行了调速控制。详细介绍了PMSM(PermanentMagnetSynchronousMotor)的矢量控制原理。最后给出PMSM单神经元自适应PID控制的仿真结果和硬件实现方法。仿真结果表明,该系统具有良好的动态性能。

Controlo directo de binário de uma máquina assíncrona trifásica

Dissertação para obtenção do grau de Mestre em Engenharia Electrotécnica Ramo de Automação e Electrónica Industrial