Since its conception in 2005, irreversible electroporation (IRE), a non-thermal tumor ablation modality, was investigated for safety and efficacy in clinical applications concerning different organs. IRE utilizes high voltage (~3kV), short duration (~100us) pulses to create transient nanoscale defects in the plasma membrane to cause cell death due to irreversible defects, osmotic imbalances and ATP loss. More recently, high-frequency irreversible electroporation (H-FIRE), which employs narrow bipolar pulses (~0.5-10us) delivered in bursts (on time ~100us), was invented to provide benefits such as the mitigation of intense muscle contractions associated with IRE-based treatments. Furthermore, H-FIRE exhibits the potential to improve lesion predictability in homogeneous and heterogeneous tissue masses. Therapeutic IRE and H-FIRE utilize source and sink electrodes inserted into or around the tumor to deliver the treatment. Prediction of the ablation size, for a set of parameters, can be achieved by the use of pre-treatment planning algorithms that calculate the induced electric field distribution in the target tissue. An electric field above a certain threshold induces cell death and parameters are tuned to ensure complete tumor coverage while sparing the nearby healthy tissue. IRE studies have shown that the underlying field is influenced by the increase in tissue conductivity due to enhanced membrane permeability, and treatment outcome can be improved when this nonlinearity is accounted for in numerical models. Since IRE pulses far exceed the time constant of the cell (~1us), the tissue response can be treated as essentially DC a static approximation can be used to predict the field distribution. Alternately, as H-FIRE pulses are on the order of the time constant of the membrane, the tissue response can no longer be treated as DC. The complexity of the H-FIRE-induced field distribution is further enhanced due to the dispersion and non-linearity in biological tissue impedance during treatment. In this dissertation, we have studied the electromagnetic fields induced in tissue during H-FIRE using several experimental and modeling techniques. In addition, we have characterized the nonlinearity and dispersion in tissue impedance during H-FIRE treatments and proposed simpler methods to predict the field distribution to enable easier translation to the clinic. / Ph. D.

Conventional methods of cancer therapy have been severely limited by inefficient delivery of therapeutic doses without incidence of harsh and toxic side effects in normal tissues. Consequently, countless new methods for early detection and drug delivery have been investigated in the area of nanoparticles and hydrogels. Although many of these methods are promising, the complex nature of cancer increases the difficultly for the development of the perfect system. Cellulose nanocrystals (CNCs) have been studied widely for a variety of applications. Despite their advantages, investigations of their abilities in the biomedical field have not been explored. The goal of this project is to delve into the potential uses of CNCs in detection, targeted drug delivery, and potentiation of irreversible electroporation (IRE)-induced cell death in folate receptor (FR)-positive cancers. To accomplish this task we have prepared stable and reproducible CNCs from wood pulp via sulfuric acid hydrolysis. Furthermore, we have functionalized the surface of these nanoparticles and conjugated them with the targeting ligand folic acid (FA) and the fluorescent imaging agent fluorescein-5\'-isothiocyanate (FITC) to create FITC-CNC-FA; CNCs have also been conjugated with doxorubicin (DOX), a potent chemotherapeutic (DOX-ALAL-CNC-FA). We have determined FITC-CNC-FA's and DOX-ALAL-CNC-FA's ability to specifically target FR-positive cancer cells in vitro; meanwhile non-targeted CNCs (FITC-CNC) were shown unable to bind to these cell types. In addition, we have investigated FITC-CNC-FA's pharmacokinetic activity in vivo. To properly model the CNC conjugate's activity in vivo, a physiologically based pharmacokinetic (PBPK) model has been constructed. We have also examined CNCs' ability to potentiate a new technique for tumor ablation, IRE. Pre-incubation with FA-conjugated CNCs (CNC-FA) have shown an increase in cytotoxicity in FR-positive cancer cells induced by IRE. In addition, CNC-FA did not potentiate IRE-induced cytotoxicity in a FR-negative cancer cell type. For a more comprehensive understanding of CNC-FA's ability to potentiate IRE induced cytotoxicity, we optimized a 3D in vitro hydrogel system. Preliminary data suggest this method of experimentation will be more realistic to in vivo studies to be completed in the future. Together, these studies showcase CNCs as efficient and effective nano-carriers in tumor detection and treatment. / Ph. D.

Emerging technologies in the healthcare industry encompass revolutionary devices or drugs that have the potential to change how healthcare will be practiced in the future. While there are several emerging healthcare technologies in the pipeline, a few key innovations are slated to be implemented clinically sooner based on their mass appeal and potential for healthcare breakthroughs. This thesis will focus on specific topics in the emerging technological fields of nanotechnology for photothermal cancer therapy, 3D printing for irreversible electroporation applications, and imageable biomaterials. While these general areas are receiving significant attention, we highlight the potential opportunities and limitations presented by our select efforts in these fields. First, in the realm of nanomedicine, we discuss the optimization and characterization of sodium thiosulfate facilitated gold nanoparticle synthesis. While many nanoparticles have been examined as agents for photothermal cancer therapy, we closely examine the structure and composition of these specific nanomaterials and discuss key findings that not only impact their future clinical use, but elucidate the importance of characterization prior to preclinical testing. Next, we examine the potential use of 3D printing to generate unprecedented multimodal medical devices for local pancreatic cancer therapy. This additive manufacturing technique offers exquisite design detail control, facilitating tools that would otherwise be difficult to fabricate by any other means. Lastly, in the field of imageable biomaterials, we demonstrate the development of composite catheters that can be visualized with near infrared imaging. This new biomaterial allows visualization with near infrared imaging, offering potentially new medical device opportunities that alleviate the use of ionizing radiation. This collective work emphasizes the need to thoroughly optimize and characterize emerging technologies prior to preclinical testing in order to facilitate rapid translation. / Master of Science

Diese Dissertation behandelt sowohl die Theorie, als auch beobachtetes Verhalten in Stoppproblemen. In einem Stoppproblem, beobachtet ein Agent die Entwicklung eines stationären, stochastischen Prozesses über die Zeit. Zu jedem Zeitpunkt genießt der Agent das Recht den Prozess zu stoppen, um eine Auszahlung einzustreichen die Funktion des gegenwärtigen und der vergangenen Realisationen des Prozesses sind. Das Ziel des Agenten ist es den Stoppzeitpunkt so zu wählen, dass die erwartete Auszahlung oder der erwartete Verlust durch Stoppen maximiert bzw. minimiert wird. Stoppprobleme dieser Art konstituieren können als die einfachsten, jedoch wirklich dynamischen Entscheidungsprobleme in der ökonomischen Theorie angesehen werden Das erste Kapitel legt neue theoretische Resultate hinsichtlich der optimalen Stoppstrategien unter Erwartungsnutzentheorie, sog. gain-loss utilities und Bedauerungspräferenzen vor. Das zweite Kapitel behandelt sodann die Ergebnisse eines Laborexperiments in dem die theoretischen Vorhersagen getestet werden. Kapitel drei beschäftigt sich mit der Situation in der die Agenten nicht vollständig über Wahrscheinlichkeiten für künftige Ereignisse informiert sind, d.h. es herrscht Ambiguität. / This dissertation is concerned with theory and behavior in stopping problems. In a stopping problem an agent or individual observes the realization of some exogenous and stationary stochastic process over time. At every point in time, she has the right or the once-only option to stop the process in order to earn a function of the past and current values of the process. The agent''s objective then is to choose the point in time to exercise the option in order to maximize an expected reward or to minimize an expected loss. Such problems constitute the most rudimentary, yet truly dynamic class of choice problems that is studied in economics. The first chapter provides new theoretical results about optimal stopping both under expected utility, as well as gain-loss utility and regret preferences. The second chapter presents a laboratory experiment that tests several of the theoretical predictions about behavior made in the first chapter. The third chapter is concerned with stopping behavior in a setting, where the probability law that drives the observed process is not perfectly known to the decision maker, i.e. there is ambiguity.

Els tractaments actuals pel càncer de pàncreas presenten un eficàcia limitada de manera que es necessari el desenvolupament de noves teràpies antitumorals. La teràpia gènica pel càncer de pàncreas basada en l’ús d’adenovirus es troba limitada per la baixa capacitat dels virus d’arribar a les masses tumoral, de distribuir-se pel tumor i d’infectar les cèl·lules tumorals. Nosaltres hem observat que l’administració intraductal d’adenovirus al ducte biliar de ratolins Ela-myc permet arribar als tumors pancreàtics de manera més eficient que per la via sistèmica. A més a més permet transduir la majoria de la massa tumoral restringint l’expressió adenoviral al teixit pancreàtic. D’altre banda, l’administració intraductal del tractament AduPARTat8TK/GCV retarda significativament el creixement tumoral i disminueix la toxicitat associada al tumor. El nou adenovirus AdTATMMP és activat per les MMP2/9 restaurant la capacitat de transducció de l’AdYTGRE in vitro, i incrementant 7,3 vegades la infecció del tumor pancreàtic. El tractament combinat de l’AduPARTat8TK/GCV amb gemcitabina presenta un efecte sinèrgic in vitro, però no millora la eficàcia antitumoral de les teràpies simples. D’altre banda el tractament de l’electroporació irreversible presenta efectes antitumorals significatius en tumors ortotòpics de la línia cel·lular BxPC-3-Luc i allarga la supervivència dels ratolins provocant una toxicitat mínima. / Novel therapies are needed to overcome the limited efficacy of current treatments in pancreatic cancer. Adenoviral gene therapy against pancreatic tumors is challenged by the limitation of viruses to reach the tumor mass, poorly distribute within the tumor and inefficiently transduce tumor cells. We show that intraductal administration of adenoviruses into the common bile duct of Ela-myc mice targets pancreatic tumors more efficiently than systemic delivery with relevant transduction of the bulk of the tumor and restricts expression to pancreatic tissue. Moreover, intraductal administration of AduPARTat8TK/GCV treatment significantly delayed tumor growth ameliorating tumor-associated toxicity. Noticeable the new generated MMP-activatable adenovirus AdTATMMP was susceptible to MMP2/9 activation, restored the transduction capacity of AdYTRGE in vitro, and increased 7.3 times tumor pancreas transduction. The multimodal treatment AduPARTat8TK/GCV and gemcitabine showed synergistic effects in vitro; however, did not enhance the antitumoral efficacy of single therapies. Interestingly, IRE treatment exhibited significant antitumor effects in BxPC-3-Luc orthotopic tumors and prolonged mice survival with minimal toxicity.

Preservation of tissue structure, morphology and biomarkers is of utmost importance for pathological examination of biopsy specimens for diagnostic and therapeutic purposes. However current methods employed to evade tissue degradation and preserve biomarkers have several shortcomings that include irreproducibility, morphological artifacts and altered biomarker antigenicity. These artifacts may affect the analysis and subsequent diagnosis of the tissue pathology. This creates need for developing improved preservation methods that reproducibly maintain tissue morphology and biomarker antigenicity and are simple, rapid and inexpensive. Experiments conducted for testing the hypothesis that cryopreservation procedures yield high quality morphology and antigenicity showed that cryopreservation maintains tissue structure, morphology and antigenicity at equivalent or better levels compared to standard freezing techniques. In order to understand the mechanisms of osmotic transport in cellular systems upon exposure to multi-component solutions that are prevalent in virtification protocols, experimental studies were undertaken using microfluidics for single cell manipulation. The experimental data yielded permeability parameters in binary and ternary solutions for MC3T3-E1 murine osteoblasts for the first time. The hydraulic conductivity (L(p)) decreased with increasing concentrations but the solute permeability either increased or decreased with increasing solution concentration. The changes in hydraulic conductivity were consistent with previously published trends and conform to a functional relationship in the form of Arrhenius type relationship between L(p) and solution concentration. Further a theoretical model was developed from principles of linear irreversible thermodynamics to simulate multi--‐‑component mass transport across membrane. The model was successfully validated by comparison with experimental data for murine osteoblasts and showed good agreement between the numerical predictions and experimental observations. The modeling approach can be used to investigate the transport mechanisms, which show that in multicomponent osmotic transport response, the dynamics is dictated by slower moving solute.

Thesis advisor: Eranthie Weerapana / The 1,3,5-triazine is a privileged scaffold in that it is planar and has three-fold symmetry which allows for controlled modification around the ring structure with various substituents. In this thesis, we report on two modular inhibitor libraries that center around a 1,3,5-triazine core scaffolding system, which have been shown to target protein disulfide isomerase A1 (PDIA1), glutaredoxin-3 (GLRX3), and 6-phosphofructo-1-kinase (PFKP). Protein disulfide isomerase A1 (PDIA1) is a thiol-disulfide oxidoreductase localized in the lumen of the endoplasmic reticulum (ER), and is an important folding catalyst and chaperone for proteins in the secretory pathway. PDIA1 contains two active-site domains (a and a’), each containing a Cys-Gly-His-Cys (CGHC) active-site motif. Here, we synthesize a targeted library o second-generation triazine-based inhibitors to optimize the potency and selectivity of our lead compound, RB-11-ca. Characterization of this targeted library afforded an optimized PDIA1 inhibitor, KSC-34, which covalently modifies C53 in the a site of PDIA1 and demonstrates time-dependent inhibition of the reductase activity of PDIA1 in vitro with a kinact/KI = 9.66 x 103 M-1s-1. Interestingly, KSC-34 treatment demonstrated that a-site inhibition led to decreased secretion of amyloidogenic antibody light chain, thus illustrating that site-selective inhibitors like KSC-34 provide useful tools for delineating the pathological role and therapeutic potential of PDIA1. In 2014, our lab first reported on RB7, a dichlorotriazine-based electrophilic small molecule which displayed extremely high reactivity and selectivity toward lysine residues in the proteome. Herein, we further on this study by investigating the unique reactivity of RB7 through the synthesis of a second-generation small molecule electrophile library and investigating proteome-wide reactivity in vitro and in situ. This library afforded KSC-46, an RB-7 analogue with p-chlorothiophenol tuning element, which provided optimal proteome reactivity to use as a scaffold for the generation of a targeted library. To take advantage of the tuned reactivity of KSC-46, a second-generation targeted library was generated to target react residues in the proteome. This library yielded two molecules, KSC-56 and KSC-65, which were identified to target glutaredoxin-3 (GLRX3) and 6-phosphofructo-1-kinase (PFKP), respectively. GLRX3 is a cytosolic, monothiol iron-sulfur cluster chaperon protein which relies on two nucleophilic cysteine residues to bind and transfer iron clusters. PFKP is known to catalyze the first irreversible step in glycolysis and regulates the flux of glucose metabolism in the cell, which makes PFKP an attract therapeutic target. KSC-56 was further characterized to bind to Cys261 in the C-terminal glutaredoxin domain of GLRX3. / Thesis (PhD) — Boston College, 2018. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Chemistry.

Neste estudo, randomizado, duplo cego, avaliamos a eficácia anestésica em quarenta e dois pacientes, do Setor de Urgência da Faculdade de Odontologia da Universidade de São Paulo, com pulpite irreversível, que receberam mepivacaína 2% (n=21) ou lidocaína 2% (n=21) associadas à epinefrina 1:100.000 para bloqueio do nervo alveolar inferior (BNAI). O sinal subjetivo de anestesia do lábio e língua, a presença de anestesia pulpar e ausência de dor durante o procedimento de pulpectomia foram avaliados, respectivamente, por indagação ao paciente, pelo testador elétrico pulpar (TEP) e pela escala de dor verbal (VAS). Pela técnica pterigomandibular indireta das três posições, foi injetado primeiramente um tubete (1,8mL), e depois de 10 minutos, foi testada a anestesia pulpar (AP), pelo TEP, por duas leituras negativas ao estímulo máximo (80A) do aparelho. Quando não instalada a AP, um segundo tubete (mais 1,8mL) era reinjetado. Confirmada a anestesia pulpar, iniciava-se a pulpectomia. O sucesso do BNAI foi definido como a capacidade de acessar a câmara pulpar e a realização da pulpectomia sem relato de dor (VAS) pelo paciente (escore 0 ou 1), enquanto o insucesso foi caracterizado pelo incomodo/dor (escore 2 ou 3), que impedisse a continuação. Nesse caso, um terceiro e último tubete foi dado por técnicas complementares (intraligamentar ou intrapulpar) para finalizar o procedimento. Na análise estatística utilizou-se o teste Exato de Fisher e ANOVA com nível de significância fixado em 5%. Obtivemos que no grupo Mepivacaína com 1,8mL, a taxa de anestesia pulpar (AP) foi de 52% (11/21), e sucesso no BNAI de 55% (6/11); a injeção de mais 1,8mL (2º tubete) aumentou a AP para 86% (18/21) e o sucesso no BNAI para 55% (10*/18). No Grupo Lidocaína, com 1.8mL, a taxa da AP foi de 33% (7/21), o BNAI foi de 0%; com mais 1,8mL (2º tubete), a AP aumentou para 67% (14/21) e sucesso no BNAI para 14% (2*/14) (*com diferença estatística onde p0,05). A mepivacaína com volume menor proporcionou, clinicamente, maior índice de anestesia pulpar e sucesso do BNAI (pulpectomia total), e permitiu chegar mais próximo da polpa quando comparada a lidocaína. Concluímos que a mepivacaína obteve melhores resultados no sucesso do BNAI para a realização da pulpectomia em molares inferiores com pulpite irreversível. / The aim of this, double blind randomized study, was to evaluate the anesthetic efficacy of inferior alveolar nerve block (IANB) using 2% mepivacaine and 2% lidocaine both associated with adrenaline 1:100,000 in molars with irreversible pulpitis. The sample of these study consisted of forty-two healthy patients diagnosed with irreversible pulpitis actively experiencing pain. For blocking the IAN was established the following protocol: injection of one cartridge (1.8 mL) by the technique pterigomandibular of three indirect positions, expected 10 minutes and electric pulp test (EPT) Vitality Scanner®-SybronEndo, USA was accomplishment. If tooth sensitivity pain persisted another cartridge (plus 1.8 mL) was given by the same technique and the same methodology was performed. The pulpectomy was continued after the confirmation of pulpal anesthesia, which was established as the lack of response at maximum stimulation (80A) of EPT. In cases where the patient reported pain during pulpectomy even confirmed the blockage, a third cartridge was given by complementary techniques (intraligamentary or intrapulpal) to complete the endodontic procedure. The anesthetic efficacy in IANB was established when the pulpectomy was performed without report of pain and without complementation. For analysis and comparison of results we used the Fisher exact statistical test and ANOVA with significance level set at 5%. Regarding the effectiveness of the anesthetic with 1,8 mL mepivacaine, determined pulpar anesthesia (PA) was 52% (11/21), and success in IANB (pulpectomy) 55% (6/11), the most injection of 1,8 mL, increased 86% (18/21) for AP and success in IANB to 55% (10*/18). In the lidocaine group, with 1.8mL, AP rate was 33% (7/21), the IANB was 0%, with a further 1.8 mL (cartridge 2) the AP increased to 67% (14/21) and success was to IANB 14 % (2*/14), (*statistically difference p 0.05). Mepivacaine with smaller volume clinically provided a higher rate of pulpal anesthesia and most successful of IANB (pulpectomy total), allowing to reach nearest dental pulp than lidocaine. Concluded that mepivacaine performed better in success of IANB (pulpectomy) that lidocaine in molars with irreversible pulpitis

No trabalho estuda-se o crescimento de trincas em carga monotônica e cíclica nos casos de materiais quase-frágeis, introduzindo uma lei de dano cíclico. Revisam-se conceitos sobre modelos coesivos, leis de carga-descarga, leis de evolução de dano e efeito de escala. É seguido o modelo coesivo irreversível proposto por Wang e Siegmund (2006). Em particular se dá ênfase aos efeitos de escala não estatísticos. O modelo de zona coesiva irreversível apresenta uma formulação de dano e considera carregamento em fadiga. Quando o tamanho estrutural é reduzido (ou as trinca se extendem), a fratura por fadiga não mais ocorre por propagação de trinca, mas sim por uma decoesão uniforme. O objetivo desde trabalho é implementar este modelo e verificar sua potencialidade na captura de efeitos de escala, comparando com experimentos e dados disponíveis na literatura. / At present work is intended to study crack growth in cyclic and monotonic loading in the case of quasi-brittle materials, introducing a damage mechanism, is reviewed concepts of cohesive models, loading-unloading laws, damage evolution laws and effect of scale. The irreversible cohesive zone model proposed by Wang e Siegmund (2006) is followed. In particular emphasizes in the not statistical size effects. The irreversible cohesive zone model, presents a damage formulation and considers fatigue loading. It is demonstrated in this study that, when the structure size is reduced (or extend cracks), the fatigue fracture no longer occurs by crack propagation, then occurs by uniform decohesion . The objetive of this work is implementing this model and verify its capability to capture the scale effect compared with experiments and data available in literature.

Este trabalho procura responder a questão: \"quais as barreiras conceituais que impedem a compreensão da segunda lei da Termodinâmica pelos estudantes? A partir de três questões apresentadas a estudantes do terceiro ano do ensino médio e do primeiro ano do curso superior de licenciatura em física, foram coletadas as respostas produzidas; posteriormente analisadas, permitiram a explicitação de concepções que, confrontadas com as idéias da Termodinâmica clássica, revelaram um raciocínio inadequado. A idéia de sistema centrado no objeto, parece constituir uma barreira conceitual difícil de transpor; assim também ocorre com os conceitos de processo e transformação exibidos pelo senso comum. Tais conceitos são fundamentais para o estabelecimento da Termodinâmica e devem ser pensados no trabalho em sala de aula capaz de produzir conflitos cognitivos com meta a uma mudança conceitual. Por outro lado, numa perspectiva histórica, foi possível identificar alguma ressonância entre as idéias dos estudantes e o desenvolvimento da concepção de calor; focalizamos especialmente as idéias de Carnot e de Clausius ao longo da elaboração das leis da Termodinâmica. O trabalho também identificou lacunas decorrentes da forma como os conteúdos científicos são apresentados na sala de aula e nos livros didáticos o que pode explicar as dificuldades encontradas. Finalmente, o trabalho apresenta subsídios de relevância pedagógica que podem contribuir para a melhoria do ensino da Termodinâmica no nível médio, com atividades especiais que tocam nos pontos conflitantes entre as concepções dos estudantes e o conhecimento científico. / This work aims to answering the following question: what are the conceptual barriers that prevent students from understanding the second law of Thermodynamics ? Starting from three questions that were proposed to third year secondary school and to first year physics teaching university students, the collected answers were analysed and led to an explicitation of conceptions that disclose inadequate reasoning when confronted with the ideas of classical Thermodynamics. The idea of a system centered on the object seems to be a conceptual barrier that is difficult to overcome and the same happens with the concepts of process and change that are usual in commonsense. But these concepts are essential for the fundamentals of Thermodynamics and should be used in class work to create cognitive conflicts aiming to a conceptual change. On the other hand, from a historical perspective, it was possible to identify some of the students ideas in the development of the concept of heat particularly in the development of the ideas of Carnot and Clausius in the elaboration of the laws of thermodynamics. The work has also identified gaps resulting from the way scientific contents are presented in the classroom and in textbooks that could explain the students difficulties. At last, the work presents pedagogically relevant contributions that may be helpful in teaching thermodynamics in secondary schools with special activities that touch points that are conflicting between students conceptions and scientific knowledge.