O fluconazol é um fármaco antifúngico utilizado na prevenção e tratamento de infecções micóticas. Atualmente, no mercado brasileiro, vários laboratórios farmacêuticos comercializam produtos a base de fluconazol na forma de cápsulas de 150 mg. Estes produtos são considerados similares e, portanto, teoricamente intercambiáveis, por conterem o mesmo princípio ativo nas mesmas dosagem e forma farmacêutica. No entanto, não existem estudos atestando a bioequivalência entre eles. Pretendeu-se, nesse trabalho, realizar avaliação biofarmacotécnica in vitro (cinética de dissolução) e in vivo (bioequivalência) de duas formulações do mercado nacional contendo fluconazol: Zoltec® 150 mg (laboratórios Pfizer Ltda.), considerado produto referência (R) e Flunazol® 150 mg (Laboratórios Sintofarma S.A.), considerado produto teste (T). Inicialmente, desenvolveu-se método para a análise da cinética de dissolução, já que não existe teste oficial de dissolução para formas farmacêuticas contendo fluconazol. Após padronização do método, avaliou-se a cinética de dissolução de cápsulas de fluconazol provenientes de dois lotes de R e dois lotes de T por meio dos parâmetros ks (constante de velocidade de dissolução) e t85% (tempo necessário para dissolução de 85% do fármaco presente na forma farmacêutica), derivados dos perfis de dissolução. Obteve-se ks de 0,1079 min-1 e 0,1377 min-1 para os dois lotes de R testados e 0,5421 min-1 para os dosi lotes de T, e t85% entre 15,09 min e 20,06 min para R e entre 5,64 min e 6,02 min para T. O ensaio de bioequivalência foi do tipo aleatório cruzado, com coleta de amostras de sangue até 96 horas após administração dos produtos R e T a 28 voluntários em jejum. Para quantificação do fluconazol em amostras de plasma desenvolveu-se e validou-se método simples, exato, preciso e sensível por cromatografia líqüida de alta eficiência (CLAE) sem utilização de padrão interno, e detecção em ultravioleta a 210 nm, após extração com solvente orgânico em meio básico. A bioequivalência entre os produtos foi determinada através da comparação dos parâmetros farmacocinéticos Cmax (concentração plasmática máxima), tmax (tempo necessário para Cmax) e AUCT (área sob a curva de decaimento plasmático) obtidos para R e T. Os resultados foram submetidos a análise estatística conforme recomendado pelo FDA-USA, determinando-se os intervalos de confiança 90% (I.C. 90%) para as relações entre Cmax de T e R e AUCT de T e R. Os valores médios de Cmax, tmax e AUCT para R e T foram, respectivamente: 3,64 mg/L e 3,75 mg/L; 2,96 h e 2,79 h; 153,33 mgh/L e 154,45 mgh/L. Os I.C. 90% para Cmax e AUCT foram, respectivamente, 101,06% a 105,45% e 97,96% a 103,36%. Concluiu-se que R e T são bioequivalentes, podendo ser administrados de forma intercambiável, sem prejuízo do efeito terapêutico. / Fluconazole is an antifungal agent widely used in the prevention and treatment of invasive fungal infections. Many brazilian pharmaceutical industries manufacture capsules containing 150 mg of fluconazole. As such products contain the same amount of the same therapeutically active ingredient in the same dosage form, they are considered to be interchangeable, indeed no bioequivalence study have been conducted to assess this. The present study was designed to perform in vitro (dissolution kinetics) and in vivo (bioequivalence) biopharmaceutical evaluation of two commercial products available in Brazil: Zoltec® 150 mg (Pfizer) as the reference product (R) and Flunazol® 150 mg (Sintofarma) as the test product (T). There is no official dissolution method for fluconazole dosage forms so initially a methos was developed and standardized for the evaluation of dissolution kinetics of fluconazole capsules. Dissolution kinetics for samples from two batches of R and two batches of T was analysed through ks (dissolution rate constant) and t85% (time for dissolution of 85% of the drug in the dosage form), obtained from dissolution profiles. Results showed ks values of 0,1079 min-1 and 0,1377 min-1 for the two tested batches of R and 0,5421 min-1 for both tested batches of T, and t85% values between 15,09 min and 20,06 min for R and between 5,64 min and 6,02 min for T. Bioequivalence assay was crossover and randomized. Blood samples were collected throughout a 96 hours period after administration of R and T to 28 fasting volunteers. A simple, accurate, precise and sensitive high-performance liquid chromatographic (HPLC) method without internal standard, and ultraviolet detection at 210 nm, was developed and validated for quantification of fluconazole im plasma samples after liquid-liquid extraction. Bioequivalence was assessed through pharmacokinetic parameters Cmax (peak plasma concentration), tmax (time to reach Cmax) and AUCT (área under the “plasma concentration vs time" curve) for R and T. Results were submitted to statistical analysis according to the FDA-USA and 90% confidence intervals (90% C.I.) were calculated for T and R Cmax ratios and T and R AUCT ratios. Average Cmax, tmax and AUCT values for R and T were, respectively: 3,64 mg/L and 3,75 mg/L; 2,96 h and 2,79 h; 153,33 mgh/L and 154,45 mgh/L. 90% C.I. for Cmax and AUCT were, respectively, 101,06% - 105,45% and 97,96% - 103,36%. Results show that R and T are bioequivalent and can be administered in an interchangeable way, without any prejudice of therapeutic effect.

Il est de plus en plus reconnu que les connexions du cerveau jouent un rôle important sur la fonction cérébrale, en particulier les fonctions cognitives supérieures comme le langage. Cependant l’imperfection des techniques traitant les connexions macroscopiques humaines a empêché l’avancement de nos connaissances sur l'anatomie des faisceaux. Nous nous appuyons ainsi aujourd’hui essentiellement sur la littérature du XIXème siècle. Les définitions des trajets et des connexions anatomiques de nombreux faisceaux sont constamment débattues. En utilisant l'imagerie de diffusion, nous réévaluons les anatomies des faisceaux clés dans une grande cohorte saine. Nous utilisons une nouvelle approche de segmentation des faisceaux qui vise à reproduire la méthode introduite par les dissectionistes. Celle-ci définit un tract comme l’ensemble des fibres passant par une même tige, minimisant ainsi l’a priori sur leurs terminaisons. Nous nous concentrons sur les faisceaux d'association des capsules externe et extrême, notamment les faisceaux occipito-frontal inférieur (FOFI) et unciné (FU) impliqués dans le circuit du langage ventral. Nous passons en revue la littérature sur ces tracts, fournissons des descriptions détaillées de leurs connectivités anatomiques et donnons un nouvel éclairage sur leur asymétrie et organisation interne. Dans une première étude, nous confirmons que les deux faisceaux ont de plus vastes projections dans le cortex qu'on ne le pensait, et nous présentons de nouveaux résultats concernant les branches asymétriques des faisceaux. Dans une deuxième étude, nous étudions en profondeur le FU et ses sous composantes. Nous résolvons un débat d’un siècle en exhibant clairement sa frontière avec le FOFI et nous identifions pour chaque sous composante des caractéristiques anatomiques distinctives y compris des asymétries. Ces résultats apportent un éclairage nouveau sur le FOFI et le FU qui sera crucial pour démêler leurs rôles multifonctionnels. / The importance of the brain’s connections for cerebral function is increasingly emphasized especially for higher cognitive functions like language. But the imperfection of the techniques used to address the human macroscopic connections has prevented the advancement of our knowledge on the anatomy of fibre pathways. Thus we rely heavily on XIXth century literature. Controversy surrounding the anatomical course and connections of many fibre pathways persists. Using diffusion imaging, we reevaluate the anatomies of key pathways in a large healthy cohort. We use a novel tract segmentation approach that aims to reproduce the method introduced by dissectionists – defining a tract as all fibers passing through a stem, thus minimizing a priori on their terminations. We focus on the association pathways of the external and extreme capsules, namely the inferior fronto-occipital (IFOF) and uncinate fasciculi (UF), implicated in the ventral language circuitry. We review the literature on these tracts, provide detailed descriptions of their connectional anatomies and present new insights regarding their asymmetry and internal organization. In a first study, we confirm that both tracts have more extensive projections within the cortex than previously thought and present new results regarding asymmetrical tract branches. In a second study we further investigate the UF including its subcomponents. We resolve a century old debate by clarifying its elusive boundary with the IFOF and reveal the distinctive anatomical features including asymmetry patterns of each subcomponent. These results shed new light on the IFOF and UF and will be crucial for disentangling their multifunctional roles.

O fluconazol é um fármaco antifúngico utilizado na prevenção e tratamento de infecções micóticas. Atualmente, no mercado brasileiro, vários laboratórios farmacêuticos comercializam produtos a base de fluconazol na forma de cápsulas de 150 mg. Estes produtos são considerados similares e, portanto, teoricamente intercambiáveis, por conterem o mesmo princípio ativo nas mesmas dosagem e forma farmacêutica. No entanto, não existem estudos atestando a bioequivalência entre eles. Pretendeu-se, nesse trabalho, realizar avaliação biofarmacotécnica in vitro (cinética de dissolução) e in vivo (bioequivalência) de duas formulações do mercado nacional contendo fluconazol: Zoltec® 150 mg (laboratórios Pfizer Ltda.), considerado produto referência (R) e Flunazol® 150 mg (Laboratórios Sintofarma S.A.), considerado produto teste (T). Inicialmente, desenvolveu-se método para a análise da cinética de dissolução, já que não existe teste oficial de dissolução para formas farmacêuticas contendo fluconazol. Após padronização do método, avaliou-se a cinética de dissolução de cápsulas de fluconazol provenientes de dois lotes de R e dois lotes de T por meio dos parâmetros ks (constante de velocidade de dissolução) e t85% (tempo necessário para dissolução de 85% do fármaco presente na forma farmacêutica), derivados dos perfis de dissolução. Obteve-se ks de 0,1079 min-1 e 0,1377 min-1 para os dois lotes de R testados e 0,5421 min-1 para os dosi lotes de T, e t85% entre 15,09 min e 20,06 min para R e entre 5,64 min e 6,02 min para T. O ensaio de bioequivalência foi do tipo aleatório cruzado, com coleta de amostras de sangue até 96 horas após administração dos produtos R e T a 28 voluntários em jejum. Para quantificação do fluconazol em amostras de plasma desenvolveu-se e validou-se método simples, exato, preciso e sensível por cromatografia líqüida de alta eficiência (CLAE) sem utilização de padrão interno, e detecção em ultravioleta a 210 nm, após extração com solvente orgânico em meio básico. A bioequivalência entre os produtos foi determinada através da comparação dos parâmetros farmacocinéticos Cmax (concentração plasmática máxima), tmax (tempo necessário para Cmax) e AUCT (área sob a curva de decaimento plasmático) obtidos para R e T. Os resultados foram submetidos a análise estatística conforme recomendado pelo FDA-USA, determinando-se os intervalos de confiança 90% (I.C. 90%) para as relações entre Cmax de T e R e AUCT de T e R. Os valores médios de Cmax, tmax e AUCT para R e T foram, respectivamente: 3,64 mg/L e 3,75 mg/L; 2,96 h e 2,79 h; 153,33 mgh/L e 154,45 mgh/L. Os I.C. 90% para Cmax e AUCT foram, respectivamente, 101,06% a 105,45% e 97,96% a 103,36%. Concluiu-se que R e T são bioequivalentes, podendo ser administrados de forma intercambiável, sem prejuízo do efeito terapêutico. / Fluconazole is an antifungal agent widely used in the prevention and treatment of invasive fungal infections. Many brazilian pharmaceutical industries manufacture capsules containing 150 mg of fluconazole. As such products contain the same amount of the same therapeutically active ingredient in the same dosage form, they are considered to be interchangeable, indeed no bioequivalence study have been conducted to assess this. The present study was designed to perform in vitro (dissolution kinetics) and in vivo (bioequivalence) biopharmaceutical evaluation of two commercial products available in Brazil: Zoltec® 150 mg (Pfizer) as the reference product (R) and Flunazol® 150 mg (Sintofarma) as the test product (T). There is no official dissolution method for fluconazole dosage forms so initially a methos was developed and standardized for the evaluation of dissolution kinetics of fluconazole capsules. Dissolution kinetics for samples from two batches of R and two batches of T was analysed through ks (dissolution rate constant) and t85% (time for dissolution of 85% of the drug in the dosage form), obtained from dissolution profiles. Results showed ks values of 0,1079 min-1 and 0,1377 min-1 for the two tested batches of R and 0,5421 min-1 for both tested batches of T, and t85% values between 15,09 min and 20,06 min for R and between 5,64 min and 6,02 min for T. Bioequivalence assay was crossover and randomized. Blood samples were collected throughout a 96 hours period after administration of R and T to 28 fasting volunteers. A simple, accurate, precise and sensitive high-performance liquid chromatographic (HPLC) method without internal standard, and ultraviolet detection at 210 nm, was developed and validated for quantification of fluconazole im plasma samples after liquid-liquid extraction. Bioequivalence was assessed through pharmacokinetic parameters Cmax (peak plasma concentration), tmax (time to reach Cmax) and AUCT (área under the “plasma concentration vs time” curve) for R and T. Results were submitted to statistical analysis according to the FDA-USA and 90% confidence intervals (90% C.I.) were calculated for T and R Cmax ratios and T and R AUCT ratios. Average Cmax, tmax and AUCT values for R and T were, respectively: 3,64 mg/L and 3,75 mg/L; 2,96 h and 2,79 h; 153,33 mgh/L and 154,45 mgh/L. 90% C.I. for Cmax and AUCT were, respectively, 101,06% - 105,45% and 97,96% - 103,36%. Results show that R and T are bioequivalent and can be administered in an interchangeable way, without any prejudice of therapeutic effect.

abstract: In recent years, conventional convolutional neural network (CNN) has achieved outstanding performance in image and speech processing applications. Unfortunately, the pooling operation in CNN ignores important spatial information which is an important attribute in many applications. The recently proposed capsule network retains spatial information and improves the capabilities of traditional CNN. It uses capsules to describe features in multiple dimensions and dynamic routing to increase the statistical stability of the network. In this work, we first use capsule network for overlapping digit recognition problem. We evaluate the performance of the network with respect to recognition accuracy, convergence and training time per epoch. We show that capsule network achieves higher accuracy when training set size is small. When training set size is larger, capsule network and conventional CNN have comparable recognition accuracy. The training time per epoch for capsule network is longer than conventional CNN because of the dynamic routing algorithm. An analysis of the GPU timing shows that adjusting the capsule structure can help decrease the time complexity of the dynamic routing algorithm significantly. Next, we design a capsule network for speech recognition, specifically, overlapping word recognition. We use both capsule network and conventional CNN to recognize 2 overlapping words in speech files created from 5 word classes. We show that capsule network achieves a considerably higher recognition accuracy (96.92%) compared to conventional CNN (85.19%). Our results show that capsule network recognizes overlapping word by recognizing each individual word in the speech. We also verify the scalability of capsule network by increasing the number of word classes from 5 to 10. Capsule network still shows a high recognition accuracy of 95.42% in case of 10 words while the accuracy of conventional CNN decreases sharply to 73.18%. / Dissertation/Thesis / Masters Thesis Electrical Engineering 2018

Francisella tularensis is the etiological agent of tularemia, a severe and potentially fatal disease in humans. It is extremely infectious by the aerosol route, being thought to cause disease in humans with an infectious dose as small as one to ten organisms, which led to its weaponization by several nations and classification as a category A select agent by the Centers for Disease Control and Prevention. An intracellular pathogen, relatively little is known about the mechanisms by which Francisella is capable of successfully modulating host cell processes to escape its phagosome and replicate within the cytosol and what genes beyond the Francisella pathogenicity island are required. Furthermore, in the context of aerosol exposure, it is unknown what cells F. tularensis initially interacts with and the overall contribution of those interactions to inhalational tularemia. I initiated this study by generating an in vitro model system to study interactions of F. tularensis with epithelial cell lines in tissue culture. Utilizing this system, I determined that F. tularensis LVS was capable of adherence to human epithelial cell lines of alveolar (A549), bronchial airway (HBE), and cervical carcinoma (HEp-2) origin. Furthermore, LVS was capable of invading these cell lines and growing productively within them. In order to detect genes important for virulence in this system, I generated a ~15,000 member transposon library in virulent strain Schu S4 that was could be screened in a high-throughput manner by transposon site hybridization. As uptake in the in vitro epithelial cell line system was relatively inefficient, I screened this library through human primary macrophages. Results of the screen implicated 207 genes as negatively selected in the human macrophage model. Of these, I generated mutants in genes residing in a locus of the Francisella chromosome, FTT1236, FTT1237, and FTT1238, to determine their virulence phenotypes. Mutants in these genes demonstrated significant vulnerability to complement-mediated lysis as compared with wild type Schu S4. Analysis of purified LPS and capsule from these mutants further showed that they had marked defects in O-antigen and capsular polysaccharide biosynthesis. Complementation of these mutants restored surface polysaccharide biosynthesis and further determined that FTT1236 and FTT1237 compose an operon, as a mutation in FTT1236 is polar onto FTT1237. Characterization of the intracellular defect of these mutants in the absence of active complement demonstrated that they were taken up more efficiently by primary human macrophages than wild type Schu S4 and were capable of phagosomal escape but exhibited reduced intracellular growth. Microscopic analysis of macrophages infected with mutant bacteria revealed that, as early as 16 hpi, these macrophages exhibited signs of cell death. In contrast, cells infected with Schu S4 exhibited a healthy, spread morphology as late as 32 h, despite significantly more extensive F. tularensis cytosolic replication. Quantitation of cell death by the release of lactate dehydrogenase, signifying membrane permeability, confirmed that mutants in FTT1236, FTT1237, and FTT1238 induced early cell death in infected macrophages as compared with wild type Schu S4. Together, this work contributes to our understanding of the factors, such as O-antigen and capsule, required for and genes involved in Francisella's lifecycle as an intracellular pathogen.

Biomedical engineering is the application of engineering principles and techniques to the medical field. It combines the design and problem solving skills of engineering with medical and biological sciences to improve healthcare diagnosis and treatment. As the result of improvements in robotics and micro technology science in the 20th century, micro electromechanical system technology has joined with medical applications which results in micro robotic medical applications. Drug delivery is one of the most important and controversial topics which scientists and engineers have tried to improve in medical applications. For diseases like cancer, localized drug delivery is a highlight target involving bombarding a small area of a human’s body and this technology has not been completely achieved yet. The ultimate objective of this thesis is the development of wireless capsule robot controlled by a magnetic drive unit. A magnetic drive unit is a system that consists of electromagnets, which produce the magnetic field from outside of the patient’s body. The capsule robot, which is the slave robot in the system, moves inside a human’s gastrointestinal tract. This project is focused mainly on a human esophagus and all the experiments are done in a prototype of the human’s esophagus. Drug delivery for diseases like cancer is the objective of the capsule robot. The proposed design consists of a slave permanent magnet for the motion of capsule robot in a tube, a reservoir of drug, and a micro mechanical mechanism for drug release. The capsule robot is fabricated and developed in a 12mm length and 5mm diameter with the weight of 1.78 grams without the built-in permanent magnet. The drug delivery system is a semi-magnetized system, which can be controlled by an external magnetic field. It consists of a mechanical plunger and spring, which can be open and close through an external magnetic field manipulation. The amount of drug for a desired location can be controlled by manipulating the external magnetic field. To achieve this target, analytical modeling is conducted. A numerical simulation and an experimental setup demonstrate that a capsule robot in a human esophagus in a simple and multi channel system. Horizontal control is set for the capsule robot, using a custom-designed controller and a colored liquid is released with the external magnetic field. The present study with its fabricated prototype is a research is this area to prove the concept of wireless control of a robot inside a human body and the potential for a drug delivery system. It is expected that the results achieved in this project will help realize and promote capsule robot for medical treatments.

China is now the main supplier in the generic pharmaceutical and bulk drugs supply market. The low-cost sourcing of raw materials from domestic manufacturers allows Chinese pharmaceutical companies to be competitive. Purchasing becomes strategic importance to the overall business performance through the implementation of concrete purchasing practices. This thesis is mainly to study the capsule purchasing practices in Chinese pharmaceutical companies. Three objectives are proposed in order to achieve the purpose: 1) Describe the capsule purchasing procedures of the companies; 2) Compare and analyse similarities and differences of their purchasing procedures; 3) Give acceptable reasons for these similarities and differences. These objectives are fulfilled on the basis of the developed van Weele’s purchasing procedure model and the factors integrated in purchasing. In this multiple-case study, qualitative approach is utilized in order to describe and interpret the how and why questions. The unstructured face-to face interviews are used. The study finds that the capsule purchasing strategy in Chinese pharmaceutical companies is at the stage of supply management and this may induct the raw material purchasing strategy in Chinese pharmaceutical industry.

Biomedical engineering is the application of engineering principles and techniques to the medical field. It combines the design and problem solving skills of engineering with medical and biological sciences to improve healthcare diagnosis and treatment. As the result of improvements in robotics and micro technology science in the 20th century, micro electromechanical system technology has joined with medical applications which results in micro robotic medical applications. Drug delivery is one of the most important and controversial topics which scientists and engineers have tried to improve in medical applications. For diseases like cancer, localized drug delivery is a highlight target involving bombarding a small area of a human’s body and this technology has not been completely achieved yet. The ultimate objective of this thesis is the development of wireless capsule robot controlled by a magnetic drive unit. A magnetic drive unit is a system that consists of electromagnets, which produce the magnetic field from outside of the patient’s body. The capsule robot, which is the slave robot in the system, moves inside a human’s gastrointestinal tract. This project is focused mainly on a human esophagus and all the experiments are done in a prototype of the human’s esophagus. Drug delivery for diseases like cancer is the objective of the capsule robot. The proposed design consists of a slave permanent magnet for the motion of capsule robot in a tube, a reservoir of drug, and a micro mechanical mechanism for drug release. The capsule robot is fabricated and developed in a 12mm length and 5mm diameter with the weight of 1.78 grams without the built-in permanent magnet. The drug delivery system is a semi-magnetized system, which can be controlled by an external magnetic field. It consists of a mechanical plunger and spring, which can be open and close through an external magnetic field manipulation. The amount of drug for a desired location can be controlled by manipulating the external magnetic field. To achieve this target, analytical modeling is conducted. A numerical simulation and an experimental setup demonstrate that a capsule robot in a human esophagus in a simple and multi channel system. Horizontal control is set for the capsule robot, using a custom-designed controller and a colored liquid is released with the external magnetic field. The present study with its fabricated prototype is a research is this area to prove the concept of wireless control of a robot inside a human body and the potential for a drug delivery system. It is expected that the results achieved in this project will help realize and promote capsule robot for medical treatments.

Reproduction de : Thèse doctorat : Pharmacie. Biochimie et biologie moléculaire : Reims : 2007. / Titre provenant de l'écran-titre. Bibliogr. f.126-143.

Salmonella infections remain one of the most common food borne diseases worldwide. Gastroenteritis, which can be caused by many non-typhoidal Salmonella (NTS) serovars, is relatively common in North America. One of the main risk factors of NTS gastroenteritis is travel to endemic areas in the developing world. The current treatment of NTS infections with antibiotics is reserved for severe cases. A growing concern with antibiotic use is that clinical isolates are becoming drug resistant. Although most NTS infections are self-limiting in nature, the burden on the body and recovery can take several months. Thus, it is vital to prevent NTS infections rather than solely rely on treatment. We have previously discovered two novel surface associated polysaccharides in Salmonella: O-Antigen capsule and X-factor. Not only O-Antigen Capsule is considered a common surface antigen, but its’ genes were found to be expressed during in vivo infections in mice. Such an antigen would be a suitable candidate in developing a vaccine against Salmonella induced gastroenteritis. The goal of this research was to evaluate the use of O-Antigen capsule to develop a traveler’s vaccine for NTS associated gastroenteritis. Results and Conclusions: We have developed a purification protocol and purified the capsule and X-factor from Salmonella Typhimurium, Enteritidis, and Heidelberg. Lipopolysaccharide (LPS) was co-isolated with O-Antigen capsule, but removed using Triton extraction. Salmonella LPS is strain-specific and an adaptive immune response against LPS will not provide cross-protection. We generated specific immune sera in rabbits to recognize O-Antigen capsule and X-factor produced by Salmonella Typhimurium and Enteritidis. We used a mouse model to determine the immunization dose of O-Antigen capsule and showed that conjugation is necessary to enhance the immune response in mice. To boost capsule production, we analyzed PyihUTSRQPO activity using a luciferase-based reporter system. Deletion of a putative transcriptional repressor (YihW) resulted in over 100-fold increase in PyihUTSRQPO confirming YihW as a repressor. We have also looked at the effect of growth media, temperature, and sugar precursors on PyihUTSRQPO activity, and were able to show that PyihUTSRQPO has highest activity in Tryptone broth at 30oC in the absence of any additional sugars.