Peritoneal carcinomatosis (PC), the intra-abdominal dissemination of malignancy, is equated with a 5-year survival of 15%, depending on the source. Appendiceal PC is a challenge to treat as cancer cells are embedded in copious amounts of mucin and are difficult to target. Oncolytic viruses (OVs) preferentially replicate and lyse cancer cells and present a targeted, novel strategy for PC. The hypothesis of this study is that appendiceal PC will show variable susceptibility to OVs and that protein expression in these tumours will predict OV replication efficiency. Human appendiceal PC infected ex-vivo with 4 different OVs displayed variable infectivity and replication by fluorescence microscopy and plaque assay. Immunohistochemistry analysis revealed differential expression of IRF3, pERK and TK in tumour compared to normal appendix. No correlation of protein expression with viral replication was observed. Personalizing OV therapy will be critical in the optimization of future care of patients treated with this modality.

A fototerapia com lasers em baixa intensidade de lesões de herpes simples tem sido demonstrada clinicamente ora prevenindo a formação de vesículas, ora cicatrizando rapidamente as lesões e até aumentando o espaço de tempo entre o aparecimento dessas manifestações recorrentes. No entanto, os mecanismos básicos de ação dos lasers nessas situações são desconhecidos. Dessa forma, o objetivo do trabalho foi realizar ensaios in vitro utilizando células epiteliais em cultivo e culturas do vírus HSV-1 para estudar a interferência do laser em baixa intensidade na infecção do HSV-1. Material e Métodos: Culturas de vírus HSV-1 e de células epiteliais de macaco (linhagem Vero) infectadas ou não infectadas, crescidas em déficit nutricional (2 % de soro fetal bovino - sfb) foram utilizadas. As irradiações foram realizadas com um laser de GaAlAs (660 e 780 nm, área focal de 3,6 mm2). Uma, duas e três irradiações com intervalos de 6 h foram realizadas. Os grupos experimentais foram: Controle: não-irradiadas; 660 nm/ 3 J/cm2 (28 s); 660 nm/ 5 J/cm2 (38 s); 780 nm/ 3 J/cm2 (19 s) e, 780 nm/ 5 J/cm2 (25 s). Os efeitos citopáticos do HSV-1 e a viabilidade celular de culturas irradiadas e controles foram analisadas em 4 condições: 1) irradiação das células epiteliais não infectadas; 2) células epiteliais irradiadas antes da infecção; 3) irradiação dos vírus antes da infecção; 4) irradiação das células previamente infectadas pelo HSV-1. A viabilidade celular foi obtida pelo teste da redução do MTT e os efeitos citopáticos por observação em microscopia de luz. Resultados: A viabilidade celular de culturas irradiadas crescidas em déficit nutricional, independentemente do número de irradiações, foi sempre significantemente menor que aquela de culturas não-irradiadas e crescidas nas condições ideais de concentração de sfb (10 %). A viabilidade celular de culturas não infectadas foi similar em todos os grupos. O número de irradiações influenciou o crescimento celular positiva e proporcionalmente ao número de irradiações, exceto para o grupo 660 nm/ 3 J/cm2. Nenhuma diferença nos efeitos citopáticos foi observada entre os grupos, independentemente do número de irradiações nas 3 condições do estudo. A viabilidade celular de todos os grupos não mudou nem pela irradiação das células nem do vírus antes da inoculação nas células. A viabilidade de células infectadas antes da irradiação foi significantemente maior que o controle quando 2 irradiações foram realizadas. Conclusão: Nas condições deste estudo a radiação laser em baixa intensidade é capaz de aumentar o crescimento de células Vero crescidas em déficit, no entanto, não o suficiente para atingir o crescimento característico dessas células crescidas nas suas condições ideais. O número de irradiações influencia o crescimento das células de forma positiva e proporcional ao número de irradiações, exceto para o parâmetro 660 nm/ 3 J/cm2. A radiação laser não altera nem a susceptibilidade das células à infecção, nem a virulência do HSV-1. No entanto, ela prolonga a viabilidade das células infectadas pelo HSV-1. Efeitos positivos da fototerapia que tem sido relatados clinicamente parecem ser devido a efeitos no hospedeiro não relacionados com a replicação viral nas células infectadas. / Purpose: The clinical effects attributed to phototherapy relative to Herpes simplex lesions have included prevention of lesion formation, speeding the healing of lesions, and decreasing the frequency of recurrent lesions. The mechanisms underlying these findings have not been established yet. The aim of this in vitro study was to analyze the effect of phototherapy on epithelial cells, on HSV-1, and on infected epithelial cells in culture. Material and Methods: Cultures of HSV-1 and infected or non-infected monkey epithelial cells (Vero cell line) grown in deficient media (2 % fetal bovine serum-fbs) were used. The laser irradiation was delivered using a GaAlAs laser (660 and 780 nm, focal spot of 3.6 mm2). One, two and three irradiations with 6 hourintervals were done. The experimental groups were: Control: non-irradiated; 660 nm/3 J/cm2 (28 sec); 660 nm/5 J/cm2 (38 sec); 780 nm/3 J/cm2 (19 sec), and 780 nm/5 J/cm2 (25 sec). The HSV-1 cytopatic effects and the cell viability of irradiated cultures and controls were analyzed in four different conditions: 1) irradiation of noninfected epithelial cells; 2) epithelial cells irradiated prior infection; 3) virus irradiated prior infection; and 4) irradiation of HSV-1 infected cells. The cell viability was assessed by the reduction of the MTT test and the cytopatic effects by the light microscopy observation. Results: The cell viability of irradiated cultures grown in nutritional deficit, independently of the irradiation numbers, was always significantly smaller than that of non-irradiated cultures grown at the ideal serum concentration condition (10 %). The cell viability of non-infected cells was similar amongst the groups. The number of irradiations influenced the cell growth positively and proportionally to the number of irradiations, except for the 660 nm/3J/cm2 group. Any variation in cytopatic effects was observed amongst the experimental groups, independently of the irradiation numbers at the 3 conditions analyzed. The cell viability of all experimental groups were not altered either by irradiation of the cells or of the virus prior infection. The viability of infected cells prior irradiation was significantly higher than that of non-irradiated cultures when 2 irradiations were done. Conclusion: The experimental conditions for this study demonstrate that the phototherapy is capable of enhancing the growth of Vero cells grown under nutritional deficit conditions, however, not enough to reach the characteristic cell growth of cells grown at the ideal serum concentration condition. The number of irradiations influences the cell growth positive and proportionally, except when the parameter 660 nm and 3 J/cm2 was used. The laser radiation does not change either the susceptibility of the Vero cell to the HSV-1 infection or the HSV-1 virulence; however, prolongs the cell viability of HSV-1 infected cells. Positive benefits of phototherapy that have been reported clinically would appear to be due to host effects unrelated to viral replication in infected cells.

Resumo: Neste trabalho apresenta-se uma estratégia de programação linear especializada para resolver, de forma eficiente, o problema de planejamento da expansão de sistemas de transmissão de energia elétrica usando o algoritmo de busca em vizinhança variável (VNS). O modelo matem'atico para este problema é um modelo de programação linear inteira mista. A solução deste modelo através de metaheurísticas, dependendo do sistema elétrico sob análise, pode exigir a solução de centenas de milhares de problemas de programação linear (PL) que exigem elevado esforço computacional e consomem a maior parte do tempo de CPU exigido pela metaheurística. A ideia fundamental deste artigo consiste em identificar soluções básicas adequadas para iniciar o processo de solução do problema de planejamento da transmissão e os inúmeros problemas de PL exigidos pela metaheurística, fazendo que os problemas de PL sejam resolvidos de forma eficiente computacionalmente. A solução básica adequada consiste na definição uma base dual factível específica para o problema que permite que novos problemas de PL resultantes das alterações de alguns parâmetros do problema de planejamento possam ser resolvidos usando um algoritmo dual simplex canalizado. A metodologia proposta foi validada e testada em um sistema de médio porte / Abstract: This paper presents a linear programming expert strategy to solve efficiently the energy transmission network expansion planning problem using the variable neighborhood search algorithm (VNS). The mathematical model used is a mixed integer linear programming problem. Solution of this model using different metaheuristics, depending on the electrical system under analysis, may require hundreds of thousands of linear programming problems (LP) which require high computational effort and consume almost all of the CPU time required by the metaheuristic. The basic idea of this paper is to identify appropriate basic solutions which help to start the process and speed up the solution of the many subsequent subproblems generated during a typical run, solving the LP problems efficiently. The starting basic solution is defined as an appropriate dual feasible basis for the specific problem, new LP problems resulting from changes in some parameters of the planning problem are solved using a bounded dual simplex algorithm. The proposed methodology was validated and tested in a system of medium size / Orientador: Rubén Augusto Romero Lázaro / Coorientador: Jose Roberto Sanches Mantovani / Banca: Anna Diva Plasencia Lotufo / Banca: Carlos Roberto Minussi / Banca: Antônio César Baleeiro Alves / Banca: Marcos Julio Rider Flores / Doutor

Natural killer (NK) cells are non-specific killer cells of the innate immune system that eliminate target cells based on discrimination between self and non-self. Activation is carefully regulated through integration of signals received through both activating and inhibitory receptors. During the course of a herpes simplex virus type-1 (HSV-1) infection, NK cells can influence host susceptibility to infection with severe infections occurring in individuals with genetic defects in the NK cell response. In response to HSV infection, NK cells are recruited to the inflammatory tissue where ensuing reciprocal interactions with accessory cells and proinflammatory cytokines induce NK cell activation, cytolytic activity, and cytokine production, contributing to innate immune response and ultimately influencing the adaptive immune response. The objective of this study was to elucidate the multiple roles of NK cells during the numerous steps in anti-HSV immune induction. Accordingly, we have demonstrated that NK cells are novel helpers that assist and influence an anti-HSV immune response via the secretion of cytokines that enhance HSV-specific CD8+ T cell effector function and cytokine production. Taken together, data from this study presented the critical importance of NK cells in mounting an essential and efficient anti-HSV immunity. The key findings of our study were: 1. In the absence of NK cells, dendritic cells have decreased capacity to prime HSV-specific T cells. 2. HSV infected NK cells can be directly activated via toll-like receptor (TLR) in a MyD88-dependent mechanism; however, interaction with HSV infected dendritic cells yields optimal NK cell activation and function (CD69 and IFNγ). 3. TRAIL-expressing NK cells eliminate antigen-bearing immature dermal DCs (CD11c+CD8α-DR5+), that migrate to draining lymphoid organs, to facilitate antigen transfer to lymphoid resident CD8α+ DC for T cell cross priming. 4. 'Helpless' CD8+ T cell function, generated in the absence of CD4+ T cells, can be partially restored to wild-type levels by NK cell supplementation. 5. Treatment of NK cells with anti-CD69 antibody results in a heightened NK activated state and augments the adaptive immune response, without increasing NK cell numbers. These findings may contribute to the potential revelation of avenues to manipulate NK cells for anti-viral therapies.

The obligate intracellular bacterium, Chlamydia trachomatis, and Herpes Simplex Virus Type-2 (HSV-2) are the leading sexually transmitted pathogens in the world. These infections are usually asymptomatic and clinically mild, but complications can be severe. Reports of dual detection of Chlamydia and HSV within the genital tracts of humans led our laboratory to develop an in vitro Chlamydia/HSV co-infection model. Little is known regarding the specific pathogenesis of Chlamydia and HSV co-infections, but HSV-super-infection of Chlamydia-infected cells caused the chlamydiae to deviate from their normal developmental cycle into a non-replicative state termed persistence, or the chlamydial stress response. Interactions between HSV envelope protein, gD with host cell junction protein, nectin-1, were enough to stimulate the departure from normal chlamydial development. Additional data also suggested that there might be differences between single infection and co-infection outcomes in vivo. Thus, two diverging hypotheses were investigated here: i) that host nectin-1 is required for normal chlamydial development; and ii) that pathogen shedding and/or disease progression in Chlamydia and HSV-2 co-infected animals will differ from that observed in singly-infected animals. Chlamydial infection of nectin-1 knockdown cell lines revealed no inhibition of chlamydial entry, but significant reductions in inclusion size and production of infectious chlamydiae. Additionally, nectin-1 knockout mice shed fewer Chlamydia compared to wild type mice. In other studies, we developed a novel in vivo Chlamydia and HSV-2 intravaginal super-infection model in BALB/c mice. Infection with Chlamydia muridarum, followed up to 9 days later by HSV-2 super-infection, both reduced HSV shedding and protected mice from HSV-induced fatal neurologic disease compared to HSV singly-infected animals. Protection is lost when: i) infected animals are no longer shedding C. muridarum; ii) when mice are inoculated with UV-inactivated C. muridarum; or iii) when viable chlamydiae are eliminated from the genital tract using antibiotics prior to HSV-2 super-infection. Altogether, we have determined that host nectin-1 is required for chlamydial development both in vitro and in vivo, and that chlamydial pre-infection protects mice from subsequent HSV infection. We predict that these observations may lead to novel approaches to prevent human infection by these two common sexually transmitted pathogens.

Le virus Herpes Simplex de type 1 (HSV-1) possède un mode d'infection particulier dit bimodal. Il peut soit se répliquer de manière active lors d'une phase dite lytique soit migrer dans les neurones et rester en latence. Il peut réactiver pour rétablir une infection lytique. Une protéine virale majeure dans la réactivation de HSV-1 est ICP0. C'est une protéine nucléaire à activité E3 ubiquitine ligase, qui possède la particularité d'induire la dégradation par le protéasome de plusieurs protéines centromériques constitutives, ce qui provoque une déstabilisation du centromère interphasique. Mon équipe a découvert une réponse cellulaire à l'instabilité centromérique, induite par la protéine ICP0, et nommée iCDR (pour interphase Centromere Damage Response.). L'objectif général de ma thèse est de déterminer les modifications structurales que subissent les centromères endommagés par ICP0 à l'origine de l'iCDR et probablement de la réactivation. J'ai pu démontrer qu'ICP0 affectait toute la structure protéique étroitement associée aux centromères durant l'interphase. Suite à ces résultats, j'ai pu démontrer, par des analyses de digestion de chromatine à la nucléase microccocale (MNAse), que l'occupation nucléosomique de la chromatine centromérique suite à l'activité d'ICP0 était affectée de façon significative. Une étude in vivo effectuée à partir de tissus nerveux provenant de souris infectées de manière latente, a permis de démontrer une co-localisation entre les génomes HSV-1 latents et les centromères. Cette co-localisation est associée à une répression transcriptionnelle du virus. Les résultats de ma thèse montrent donc que les effets d'ICP0 sur la déstabilisation des centromères sont en relation avec un rôle de ces centromères durant la latence. Ceci suggère fortement une implication de la déstabilisation des centromères dans le processus de réactivation contrôlé par ICP0.

Esta investigación explora las relaciones entre la enfermedad dermatológica y las variables de personalidad. Se recoge una muestra de 108 pacientes de enfermedades de piel consideradas influenciables por variables psicológicas. Las enfermedades estudiadas fueron neurodermitis, psoriasis y boca urente. La enfermedad de neurodermitis se produce por rascado intenso en localizaciones accesibles al paciente y produce liquenificación de la piel en respuesta a ese rascado, frecuentemente asociado a situaciones emocionales estresantes. Como consecuencia se produce picor intenso que mantiene el ciclo de rascado, provocando unas lesiones características crónicas. Se comparan los perfiles de personalidad en estas enfermedades, con la población normal para obtener las características de personalidad en este grupo dermatológico. Los resultados avalan la hipótesis de que existen estilos de personalidad en la grupo dermatológico analizado que se diferencian de la población normal. Estas diferencias son interpretadas a la luz de la teoría de la personalidad de Millon. / This research explores the relationships between dermatologic disease and personality. 108 dermatologic patients affected with skin diseases influenced by psychological variables were recruited. The dermatologic diseases were Lichen simplex Chronicus, Psoriasis and Burning Mouth syndrome. Lichen Simplex Chronicus is produced by a intense scratching in locations accessible to the patient witch produces lichenification of the skin in response to the scratching, usually associated with emotionally stressful situations. As a consequence, itching is produced and maintains the scratching cycle witch provokes the lesions. The personality profiles of these dermatologic conditions are compared with those in normal population to obtain the personality characteristics of this dermatologic group.Results support the hypothesis that different personality styles exist between dermatologic and normal group. These differences are interpreted following the Millon theory of Personality.

This thesis aims to study the theoretical complexity and empirical performance of decomposition algorithms. We focus on linear programs with a block-angular structure. Decomposition algorithms used to be the only way to solve large-scale special structured problems, in terms of memory limit and CPU time. However, with the advances in computer technology over the past few decades, many large-scale problems can now be solved simply by using some general purpose LP software, without exploiting the problems' inner structures. A question arises naturally, should we solve a structured problem with decomposition, or directly solve it as a whole? We try to understand how a problem's characteristics influence its computational performance, and compare the relative efficiency of algorithms with and without decomposition. Two comparisons are conducted in our research: first, the Dantzig-Wolfe decomposition method (DW) versus the simplex method (simplex); second, the analytic center cutting plane method (ACCPM) versus the interior point method (IPM). These comparisons fall into the two main solution approaches in linear programming: simplex-based algorithms and IPM-based algorithms. Motivated by our observations of ACCPM and DW decomposition, we devise a hybrid algorithm combining ACCPM and DW, which are the counterparts of IPM and simplex in the decomposition framework, to take the advantages of both: the quick convergence rate of IPM-based methods, as well as the accuracy of simplex-based algorithms. A large set of 316 instances is incorporated in our experiments, so that different dimensioned problems with primal or dual block-angular structures are covered to test our conclusions.

This thesis aims to study the theoretical complexity and empirical performance of decomposition algorithms. We focus on linear programs with a block-angular structure. Decomposition algorithms used to be the only way to solve large-scale special structured problems, in terms of memory limit and CPU time. However, with the advances in computer technology over the past few decades, many large-scale problems can now be solved simply by using some general purpose LP software, without exploiting the problems' inner structures. A question arises naturally, should we solve a structured problem with decomposition, or directly solve it as a whole? We try to understand how a problem's characteristics influence its computational performance, and compare the relative efficiency of algorithms with and without decomposition. Two comparisons are conducted in our research: first, the Dantzig-Wolfe decomposition method (DW) versus the simplex method (simplex); second, the analytic center cutting plane method (ACCPM) versus the interior point method (IPM). These comparisons fall into the two main solution approaches in linear programming: simplex-based algorithms and IPM-based algorithms. Motivated by our observations of ACCPM and DW decomposition, we devise a hybrid algorithm combining ACCPM and DW, which are the counterparts of IPM and simplex in the decomposition framework, to take the advantages of both: the quick convergence rate of IPM-based methods, as well as the accuracy of simplex-based algorithms. A large set of 316 instances is incorporated in our experiments, so that different dimensioned problems with primal or dual block-angular structures are covered to test our conclusions.

Bivariate or multivariate correlated data may be collected on a sample of unit in many applications. When the experimenters concern about the failure times of two related subjects for example paired organs or two chronic diseases, the bivariate binary data is often acquired. This type of data consists of a observation point x and indicators which represent whether the failure times happened before or after the observation point. In this work, the observed bivariate data can be written with the following form {x, £_1=I(X1≤ x), £_2=I(X2≤ x)}.The corresponding optimal design problems for parameter estimation under this type of bivariate data are discussed. For this kind of the multivariate responses with explanatory variables, their marginal distributions may be from different distributions. Copula model is a way to formulate the relationship of these responses, and the association between pairs of responses. Copula models for bivariate binary data are considered useful in practice due to its flexibility. In this dissertation for bivariate binary data, the marginal functions are assumed from exponential or Weibull distributions and two assumptions, independent or correlated, about the joint function between variables are considered. When the bivariate binary data is assumed correlated, the Clayton copula model is used as the joint cumulative distribution function. There are few works addressed the optimal design problems for bivariate binary data with copula models. The D-optimal designs aim at minimizing the volume of the confidence ellipsoid for estimating unknown parameters including the association parameter in bivariate copula models. They are used to determine the best observation points. Moreover, the Ds-optimal designs are mainly used for estimation of the important association parameter in Clayton model. The D- and Ds-optimal designs for the above copula model are found through the general equivalence theorem with numerical algorithm. Under different model assumptions, it is observed that the number of support points for D-optimal designs is at most as the number of model parameters for the numerical results. When the difference between the marginal distributions and the association are significant, the association becomes an influential factor which makes the number of supports gets larger. The performances of estimation based on optimal designs are reasonably well by simulation studies. In survival experiments, the experimenter customarily takes trials at some specific points such as the position of the 25, 50 and 75 percentile of distributions. Hence, we consider the design efficiencies when the design points for trials are at three or four particular percentiles. Although it is common in practice to take trials at several quantile positions, the allocations of the proportion of sample size also have great influence on the experimental results. To use a locally optimal design in practice, the prior information for models or parameters are needed. In case there is not enough prior knowledge about the models or parameters, it would be more flexible to use sequential experiments to obtain information in several stages. Hence with robustness consideration, a sequential procedure is proposed by combining D- and Ds-optimal designs under independent or correlated distribution in different stages of the experiment. The simulation results based on the sequential procedure are compared with those by the one step procedures. When the optimal designs obtained from an incorrect prior parameter values or distributions, those results may have poor efficiencies. The sample mean of estimators and corresponding optimal designs obtained from sequential procedure are close to the true values and the corresponding efficiencies are close to 1. Huster (1989) analyzed the corresponding modeling problems for the paired survival data and applied to the Diabetic Retinopathy Study. Huster (1989) considered the exponential and Weibull distributions as possible marginal distributions and the Clayton model as the joint function for the Diabetic Retinopathy data. This data was conducted by the National Eye Institute to assess the effectiveness of laser photocoagulation in delaying the onset of blindness in patients with diabetic retinopathy. This study can be viewed as a prior experiment and provide the experimenter some useful guidelines for collecting data in future studies. As an application with Diabetic Retinopathy Study, we develop optimal designs to collect suitable data and information for estimating the unknown model parameters. In the second part of this work, the optimal design problems for parameter estimations are considered for the type of proportional data. The nonlinear model, based on Jorgensen (1997) and named the dispersion model, provides a flexible class of non-normal distributions and is considered in this research. It can be applied in binary or count responses, as well as proportional outcomes. For continuous proportional data where responses are confined within the interval (0,1), the simplex dispersion model is considered here. D-optimal designs obtained through the corresponding equivalence theorem and the numerical results are presented. In the development of classical optimal design theory, weighted polynomial regression models with variance functions which depend on the explanatory variable have played an important role. The problem of constructing locally D-optimal designs for simplex dispersion model can be viewed as a weighted polynomial regression model with specific variance function. Due to the complex form of the weight function in the information matrix is considered as a rational function, an approximation of the weight function and the corresponding optimal designs are obtained with different parameters. These optimal designs are compared with those using the original weight function.